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Motshosi P, Phinius BB, Jongman M, Baruti K, Bhebhe L, Mulenga G, Choga WT, Moyo S, Gaseitsiwe S, Anderson M. Optimization of dried blood spot for hepatitis B virus surface antibody quantification. PLoS One 2025; 20:e0304931. [PMID: 40424262 PMCID: PMC12111668 DOI: 10.1371/journal.pone.0304931] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/21/2024] [Accepted: 10/06/2024] [Indexed: 05/29/2025] Open
Abstract
Dried blood spot (DBS) cards can be used as an alternative sample collection method to plasma, however, there is no optimized elution protocol for DBS cards specifically for hepatitis B surface antibody (anti-HBs) testing. The study aimed to develop a DBS elution protocol for anti-HBs quantification. Our study sought to determine the ideal phosphate-buffered saline (PBS) volume to use by comparing three PBS volumes (300 µL, 450 µL, and 500 µL), and the optimal time to agitate DBS discs on a plate shaker (1, 2, 3, and 4 hours) to yield DBS anti-HBs concentrations that are comparable to corresponding plasma anti-HBs concentrations. The optimal DBS storage temperature (25°C, -20°C, and -80°C) was investigated to determine the ideal long-term storage temperature of the cards. Residual samples were used for optimization (2019-2021). A total of 50 DBS-plasma pairs were used throughout the study, with plasma anti-HBs concentrations being used as the golden standard to compare. A two-way analysis of variance (ANOVA) was also performed to determine the impact of PBS elution volumes, elution time, and storage temperature on the anti-HBs concentration of DBS samples. No statistically significant difference between the DBS-plasma anti-HBs pairs was observed when using 450 or 500 µL of PBS and when samples were agitated for 3 hours (p = 0.59, p = 0.50) respectively. The optimal storage temperature for DBS cards was 25°C because the results showed no statistically significant difference between DBS-plasma anti-HBs titers (p = 0.59). The two-way ANOVA analysis showed that elution volumes and time had no statistically significant impact on the DBS anti-HBs concentrations, p = 0.95 and p = 0.38 respectively. Storage temperature had a statistically significant impact on the DBS anti-HBs concentrations, p = 0.002. The optimized DBS elution protocol for anti-HBs quantification will help monitor vaccine efficacy in infants due to the low sample volumes required compared to plasma and also can be used for anti-HBs testing in resource-limited areas around the country.
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Affiliation(s)
| | - Bonolo B. Phinius
- Botswana Harvard Health Partnership, Gaborone, Botswana
- School of Allied Health Professions, Faculty of Health Sciences, University of Botswana Gaborone, Botswana
| | | | - Kabo Baruti
- Botswana Harvard Health Partnership, Gaborone, Botswana
- Department of Biological Sciences, University of Botswana, Gaborone, Botswana
| | | | - Graceful Mulenga
- Faculty of Mathematics and Statistical Sciences, Botswana International University of Science and Technology, Palapye, Botswana
| | - Wonderful T. Choga
- Botswana Harvard Health Partnership, Gaborone, Botswana
- School of Allied Health Professions, Faculty of Health Sciences, University of Botswana Gaborone, Botswana
| | - Sikhulile Moyo
- Botswana Harvard Health Partnership, Gaborone, Botswana
- School of Allied Health Professions, Faculty of Health Sciences, University of Botswana Gaborone, Botswana
- Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, United States of America
- School of Health Systems and Public Health, University of Pretoria, South Africa
- Department of Pathology, Division of Medical Virology, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg, South Africa
| | - Simani Gaseitsiwe
- Botswana Harvard Health Partnership, Gaborone, Botswana
- Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, United States of America
| | - Motswedi Anderson
- Botswana Harvard Health Partnership, Gaborone, Botswana
- Africa Health Research Institute, Durban, South Africa
- The Francis Crick Institute, London
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Ko K, Lokteva LM, Akuffo GA, Phyo Z, Chhoung C, Bunthen E, Ouoba S, Sugiyama A, Akita T, Rattana K, Vichit O, Takahashi K, Tanaka J. A comparative study of extraction free detection of HBV DNA using sodium dodecyl sulfate, N-lauroylsarcosine sodium salt, and sodium dodecyl benzene sulfonate. Sci Rep 2024; 14:25442. [PMID: 39455809 PMCID: PMC11512040 DOI: 10.1038/s41598-024-75944-7] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/20/2024] [Accepted: 10/09/2024] [Indexed: 10/28/2024] Open
Abstract
This study aimed to develop an extraction-free method for quantitative and qualitative detection of HBV DNA compared to traditional nucleic acid extraction. Paired serum and dried blood spot (DBS) samples from 67 HBsAg-positive and 67 HBsAg-negative individuals were included. Two samples with known HBV DNA titers (~ 109 copies/mL) were examined by extraction-free detection using three surfactants (0.2 to 1% of Sodium dodecyl sulfate:SDS, N-Lauroylsarcosine sodium salt:NL, Sodium dodecyl benzene sulfonate:SDBS), two stabilizing agents (0.1 or 0.01% 2-Mercaptoethanol:2ME and 3.5 or 7% Bovine Serum Albumin:BSA) and two Taq polymerases (Fast Advanced and Prime Direct Probe). HBV DNA in all 67 HBsAg-positive and 67 HBsAg-negative serum and DBS samples was directly quantified by Rt-PCR using 0.4% SDS or 0.4% NL with Fast Advance or Prime Direct Probe Taq. Extraction-free amplification was also performed. Detection limits were varied by different surfactants and Taq. SDS combined with Fast Advanced Taq showed lower detection limits, while SDS with Prime Direct Probe Taq outperformed NL or SDBS-based detection. Adding 2ME to SDS improved detection limit with Prime Direct Probe Taq but not significantly compared to SDS alone. BSA did not significantly enhance detection limits but provided insights into sample stability. The senitivity and specificity of 0.4% SDS and NL in combination with either Fast advanced or Prime Direct Probe Taq polymerase in serum samples were > 90% and 100% resepctively, while it was > 80% and 100% respectively in DBS samples. Extraction-free HBV DNA amplification provided 100% identity with original genomes. Our study suggests that SDS, NL or SDBS-based extraction-free HBV DNA detection strategies using Prime Direct Probe Taq have potential to simplify and accelerate HBV DNA detection with high sensitivity and specificity in both serum and DBS samples, with implications for resource-limited settings and clinical applications. Utilizing surfactants with 2ME is optional, and further research and validation are necessary to broaden its application in real-world diagnostics.
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Affiliation(s)
- Ko Ko
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan
| | - Lyubov Mikhailovna Lokteva
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Research Institute of Virology, Tashkent, Uzbekistan
| | - Golda Ataa Akuffo
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan
| | - Zayar Phyo
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan
| | - Chanroth Chhoung
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan
| | - E Bunthen
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- National Payment Certification Agency, National Social Protection Council, Ministry of Economic and Finance, Phnom Penh, Cambodia
| | - Serge Ouoba
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Unité de Recherche Clinique de Nanoro (URCN), Institut de Recherche en Science de La Santé (IRSS), Nanoro, Burkina Faso
| | - Aya Sugiyama
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan
| | - Tomoyuki Akita
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan
| | - Kim Rattana
- National Maternal and Child Health Center (NMCHC), Ministry of Health, Phnom Penh, Cambodia
| | - Ork Vichit
- National Immunization Program (NIP), Ministry of Health, Phnom Penh, Cambodia
| | - Kazuaki Takahashi
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan
| | - Junko Tanaka
- Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami, Hiroshima, 734-8551, Japan.
- Project Research Center for Epidemiology and Prevention of Viral Hepatitis and Hepatocellular Carcinoma, Hiroshima University, Hiroshima, Japan.
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Motshosi P, Phinius BB, Jongman M, Baruti K, Bhebhe L, Choga WT, Moyo S, Gaseitsiwe S, Anderson M. OPTIMIZATION OF DRIED BLOOD SPOT ASSAYS FOR HEPATITIS B VIRUS SURFACE ANTIBODY QUANTIFICATION. BIORXIV : THE PREPRINT SERVER FOR BIOLOGY 2024:2024.05.23.595500. [PMID: 38826424 PMCID: PMC11142191 DOI: 10.1101/2024.05.23.595500] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/04/2024]
Abstract
Dried blood spot (DBS) cards can be used as an alternative sample collection method to plasma, however, there is no optimized elution protocol for DBS cards specifically for hepatitis B surface antibody (anti-HBs) testing. The study aimed to develop a DBS elution protocol for anti-HBs quantification. Our study sought to determine the ideal phosphate-buffered saline (PBS) buffer volume to use by comparing three PBS volumes (300uL, 450uL, and 500uL), and the optimal time to agitate DBS discs on a plate shaker (1hr, 2hrs, 3hrs, and 4hrs) to yield DBS anti-HBs concentrations that are comparable to corresponding plasma anti-HBs concentrations. The optimal DBS storage temperature (25°C, -20°C, and -80°C) was investigated to determine the ideal long-term storage temperature of the cards. Residual samples were used for optimization (2019-2021). A total of 50 DBS-plasma pairs was used throughout the study, with plasma anti-HBs concentrations being used as the golden standard to compare. The analysis of results was carried out by determining the p-values of the Wilcoxon sign rank. A two-way analysis of variance (ANOVA) was also performed to determine the impact of PBS elution volumes, elution time, and storage temperature on the anti-HBs concentration of DBS samples on STATA Version 15.0. No statistically significant difference between the DBS-plasma anti-HBs pairs was observed when using 450 or 500uL of PBS buffer and when samples were agitated for 3 hours (p=0.594, p=0.499 respectively). The optimal storage temperature for DBS cards was 25°C because the results showed no statistically significant difference between DBS-plasma anti-HBs titers (p=0.594). The two-way ANOVA analysis showed that elution volumes and time had no statistically significant impact on the DBS anti-HBs concentrations, p=0.948 and p=0.381 respectively. Storage temperature had a statistically significant impact on the DBS anti-HBs concentrations, p=0.002. The optimized DBS elution protocol for anti-HBs quantification will help monitor vaccine efficacy in infants due to the low sample volumes required compared to plasma and also can be used for anti-HBs testing in resource-limited areas around the country.
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Affiliation(s)
| | - Bonolo B. Phinius
- Botswana Harvard Health Partnership, Gaborone, Botswana
- School of Allied Health Professions, Faculty of Health Sciences, University of Botswana Gaborone, Botswana
| | | | - Kabo Baruti
- Botswana Harvard Health Partnership, Gaborone, Botswana
- Department of Biological Sciences, University of Botswana, Gaborone, Botswana
| | | | - Wonderful T. Choga
- Botswana Harvard Health Partnership, Gaborone, Botswana
- School of Allied Health Professions, Faculty of Health Sciences, University of Botswana Gaborone, Botswana
| | - Sikhulile Moyo
- Botswana Harvard Health Partnership, Gaborone, Botswana
- School of Allied Health Professions, Faculty of Health Sciences, University of Botswana Gaborone, Botswana
- Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, USA
- School of Health Systems and Public Health, University of Pretoria, South Africa
- Division of Medical Virology, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg, South Africa
| | - Simani Gaseitsiwe
- Botswana Harvard Health Partnership, Gaborone, Botswana
- Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, USA
| | - Motswedi Anderson
- Botswana Harvard Health Partnership, Gaborone, Botswana
- University of KwaZulu Natal
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Virtudazo MCC, Aquino JB, Arellano RNB, Fortes RA, Kaw RC, Tantengco OAG. The role of dried blood spot tests in the detection of hepatitis B infection: A systematic review. J Viral Hepat 2024; 31:35-46. [PMID: 37789709 DOI: 10.1111/jvh.13890] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/18/2023] [Accepted: 09/23/2023] [Indexed: 10/05/2023]
Abstract
Hepatitis B remains a public health problem worldwide despite vaccine availability. Although the existing diagnostic tools help detect the infection, logistics support and limited resources and technologies affect their usefulness and reliability in developing countries. This systematic review evaluated the performance of dried blood spots (DBS) as a collection and storage tool for diagnosing an hepatitis B virus (HBV) infection. A comprehensive search using OVID, Scopus and CINAHL databases was performed to collate articles published up to April 2023 that detected Hepatitis B infections using DBS. Five reviewers independently performed identification, screening, quality assessment and data extraction. A qualitative synthesis of the included studies was conducted. Of the 402 articles, 78 met the inclusion criteria. The results show that most studies focused on populations with known HBV, HCV and/or HIV status. Approximately half (49%) of the included studies utilized the Whatman Protein Saver Card for DBS collection. The DBS samples were then predominantly stored in room temperature conditions. In line with this, storage conditions influenced the concentration and stability of the analyte from the DBS samples, affecting the accuracy of downstream diagnostic methods. ELISA methods, using hepatitis B surface antigen (HBsAg) as an HBV marker, were the most widely used diagnostic tool for detecting HBV infection in DBS samples. The simplicity and cost-effectiveness of the ELISA technique highlight its potential to be used in low-resource settings. In line with this, the detection of HBsAg using an ELISA immunoassay had higher sensitivity (85.6%-100%), and specificity (95%-100%) ranges as compared to other target molecules and methods. Although this review only performed a qualitative analysis, DBS offers a promising method for collecting and storing blood samples; however, the standardization of sampling, storing conditions and diagnostic techniques is required to ensure sustainable application.
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Affiliation(s)
| | - Jericho B Aquino
- Department of Biology, College of Arts and Sciences, University of the Philippines Manila, Manila, Philippines
| | - Rose Nicole B Arellano
- Department of Biology, College of Arts and Sciences, University of the Philippines Manila, Manila, Philippines
| | - Robert A Fortes
- Department of Biology, College of Arts and Sciences, University of the Philippines Manila, Manila, Philippines
| | - Raphaela C Kaw
- Department of Biology, College of Arts and Sciences, University of the Philippines Manila, Manila, Philippines
| | - Ourlad Alzeus G Tantengco
- Department of Biology, College of Arts and Sciences, University of the Philippines Manila, Manila, Philippines
- Department of Physiology, College of Medicine, University of the Philippines Manila, Manila, Philippines
- Department of Biology, College of Science, De La Salle University, Manila, Philippines
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Tu T, Ajoyan H, George J. Novel Assays to Solve the Clinical and Scientific Challenges of Chronic Hepatitis B. Clin Liver Dis 2023; 27:837-855. [PMID: 37778773 DOI: 10.1016/j.cld.2023.05.002] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/03/2023]
Abstract
Chronic infection with Hepatitis B is a common, incurable, and deadly infection. Despite inexpensive laboratory tests for diagnosis and management that have been established for decades, the worldwide rate of diagnosis is only ∼10%, and ∼5% of people are under treatment. Novel assays have been developed to improve linkage to care by providing more flexible approaches to determine a patient's health status. Other assays have been established to better investigate intrahepatic host-virus interactions to support clinical trials for cure research. This review outlines the clinical and scientific challenges still to be solved and the upcoming methods used to address them.
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Affiliation(s)
- Thomas Tu
- Storr Liver Centre, The Westmead Institute for Medical Research, The University of Sydney at Westmead Hospital, Westmead, New South Wales, Australia; Centre for Infectious Diseases and Microbiology, Sydney Infectious Diseases Institute, The University of Sydney at Westmead Hospital, Westmead, New South Wales, Australia.
| | - Harout Ajoyan
- Storr Liver Centre, The Westmead Institute for Medical Research, The University of Sydney at Westmead Hospital, Westmead, New South Wales, Australia
| | - Jacob George
- Storr Liver Centre, The Westmead Institute for Medical Research, The University of Sydney at Westmead Hospital, Westmead, New South Wales, Australia
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Djaogol T, Périères L, Marcellin F, Diouf A, Carrieri MP, Diallo A, Boyer S. Hepatitis B prevention and treatment needs in women in Senegal (ANRS 12356 AmBASS survey). BMC Public Health 2023; 23:825. [PMID: 37143029 PMCID: PMC10161542 DOI: 10.1186/s12889-023-15710-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2022] [Accepted: 04/19/2023] [Indexed: 05/06/2023] Open
Abstract
BACKGROUND Although mother-to-child transmission (MTCT) of hepatitis B virus (HBV) is prevalent in West Africa, epidemiological data on HBV infection in women remain scarce. We studied i) hepatitis B surface antigen (HBsAg) prevalence and its correlates, ii) HBV screening history and serological status awareness, iii) MTCT risk and treatment needs in Senegalese women. METHODS A cross-sectional population-based serosurvey for HBsAg positivity was conducted in 2018-2019 in the rural area of Niakhar (Fatick region, Senegal). Participants were offered home-based HBV screening and answered face-to-face questionnaires. HBsAg-positive participants underwent clinical and biological assessments. Data were weighted and calibrated to be representative of the area's population. Logistic regression models helped identify factors associated with HBsAg-positivity in adult women (> 15 years old). RESULTS HBsAg prevalence in adult women was 9.2% [95% confidence interval: 7.0-11.4]. Factors associated with HBsAg-positivity were being 15-49 years old (ref: ≥ 50), living in a household with > 2 other HBsAg-positive members, and knowing someone with liver disease. Only 1.6% of women had already been tested for HBV; no one who tested HBsAg positive was already aware of their serological status. In women 15-49 years old, 5% risked MTCT and none were eligible for long-term antiviral treatment. CONCLUSIONS Adult women have a high HBsAg prevalence but a low MTCT risk. Low rates of HBV screening and serological status awareness argue for the adoption of systematic screening during pregnancy using free and rapid diagnostic tests. Additionally, screening household members of HBsAg-positive women may greatly improve the cascade of care in rural Senegal. TRIAL REGISTRATION ClinicalTrials.gov identifier (NCT number): NCT03215732.
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Affiliation(s)
- Tchadine Djaogol
- Aix Marseille Univ, Inserm, IRD, SESSTIM, Sciences Economiques & Sociales de La Santé & Traitement de L'Information Médicale, ISSPAM, Marseille, France
- Univ. Bordeaux, INSERM, Institut Bergonié, BPH, U1219, CIC-P 1401, F-33000, Bordeaux, France
| | - Lauren Périères
- Aix Marseille Univ, Inserm, IRD, SESSTIM, Sciences Economiques & Sociales de La Santé & Traitement de L'Information Médicale, ISSPAM, Marseille, France
- VITROME, Campus IRD-UCAD, Dakar, Senegal
| | - Fabienne Marcellin
- Aix Marseille Univ, Inserm, IRD, SESSTIM, Sciences Economiques & Sociales de La Santé & Traitement de L'Information Médicale, ISSPAM, Marseille, France
| | | | - Maria Patrizia Carrieri
- Aix Marseille Univ, Inserm, IRD, SESSTIM, Sciences Economiques & Sociales de La Santé & Traitement de L'Information Médicale, ISSPAM, Marseille, France
| | | | - Sylvie Boyer
- Aix Marseille Univ, Inserm, IRD, SESSTIM, Sciences Economiques & Sociales de La Santé & Traitement de L'Information Médicale, ISSPAM, Marseille, France.
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Auma E, Hall T, Chopra S, Bilton S, Ramkhelawon L, Amini F, Calvert A, Amirthalingam G, Jones CE, Andrews N, Heath PT, Le Doare K. Using Dried Blood Spots for a Sero-Surveillance Study of Maternally Derived Antibody against Group B Streptococcus. Vaccines (Basel) 2023; 11:vaccines11020357. [PMID: 36851236 PMCID: PMC9966576 DOI: 10.3390/vaccines11020357] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2022] [Revised: 01/18/2023] [Accepted: 01/27/2023] [Indexed: 02/08/2023] Open
Abstract
Vaccination during pregnancy could protect women and their infants from invasive Group B Streptococcus (GBS) disease. To understand if neonatal dried blood spots (DBS) can be used to determine the amount of maternally derived antibody that protects infants against invasive GBS disease, a retrospective case-control study was conducted in England between 1 April 2014 and 30 April 2015. The DBS of cases with invasive GBS disease (n = 61) were matched with healthy controls (n = 125). The haematocrit, DBS storage temperature, freeze-thaw cycle, and paired serum/DBS studies were set up to optimise the antibody assessment. The samples were analysed using a multiplex immunoassay, and the results were assessed using parametric and nonparametric tests. Antibody concentrations were stable at haematocrits of up to 50% but declined at 75%. DBS storage at room temperature was stable for three months compared with storage from collection at -20 °C and rapidly degraded thereafter. Total IgG levels measured in DBS and paired serum showed a good correlation (r2 = 0.99). However, due to suboptimal storage conditions, no difference was found in the GBS IgG levels between DBS samples from cases and controls. We have demonstrated a proof of concept that assays utilising DBS for assessing GBS serotype-specific antibodies in infants is viable. This method could be used to facilitate future large sero-correlate studies, but DBS samples must be stored at -20 °C for long term preservation of antibody.
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Affiliation(s)
- Erick Auma
- Department of Biology, Université Claude Bernard Lyon, ENS de Lyon, CNRS, UMR, 69100 Villeurbanne, France
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
| | - Tom Hall
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
- Correspondence:
| | - Simran Chopra
- Immunity and Infection, Faculty of Medicine, Imperial College London, London SW7 2BX, UK
| | - Sam Bilton
- Neonatal Unit, John Radcliffe Hospital, Oxford University Hospitals NHS Foundation Trust, Oxford OX3 9DU, UK
| | - Laxmee Ramkhelawon
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
| | - Fahimah Amini
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
| | - Anna Calvert
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
| | - Gayatri Amirthalingam
- Immunisation and Vaccine Preventable Diseases Division, UK Health Security Agency, London NW9 5EQ, UK
| | - Christine E. Jones
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
- Faculty of Medicine and Institute for Life Sciences, University of Southampton, Southampton, SO16 6YD, UK
- NIHR Southampton Clinical Research Facility and Biomedical Research Centre, University Hospital Southampton NHS Foundation Trust, Southampton SO16 6YD, UK
| | - Nick Andrews
- Immunisation and Vaccine Preventable Diseases Division, UK Health Security Agency, London NW9 5EQ, UK
| | - Paul T. Heath
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
| | - Kirsty Le Doare
- Centre for Neonatal and Paediatric Infection, Institute for Infection and Immunity, St George’s, University of London, London SW17 0RE, UK
- Makerere University—Johns Hopkins University Research Collaboration, Kampala P.O. Box 23491, Uganda
- Pathogen Immunology Group, UK Health Security Agency, Porton Down, Salisbury SP4 0JG, UK
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Kouamé DR, Aboli RA, Kabran M, Adiko AC, Coulibaly M, Dembele B, Inwoley A. Validation of dried blood spots samples for the screening of hepatitis B virus surface by enzyme immunoassay method. J Immunol Methods 2023; 513:113412. [PMID: 36586510 DOI: 10.1016/j.jim.2022.113412] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/14/2021] [Revised: 12/26/2022] [Accepted: 12/27/2022] [Indexed: 12/29/2022]
Abstract
Dried Blood Spots (DBS) are blood collection carriers that facilitate the storage and transport of samples. Used for quality control during sero-epidemiological investigations, DBS eluate are not the conventional specimen indicated by manufacturers for enzyme immunoassay method (EIA) for hepatitis B virus surface (HBs antigen). The aim of our study was to evaluate DBS eluates used as a matrix for EIA of HBs antigen in a reference laboratory. This study took place from August 2016 to November 2017 at the Centre for Diagnosis and Research on AIDS and other infectious diseases (CeDReS) in Abidjan, Côte d'Ivoire. We used a panel of 149 whole blood samples from blood donors. The DBS performed with these samples were analyzed after elution with the HBsAg (version ULTRA) ELISA, Dia.Pro Diagnostic Bioprobes S.R.L., Sesto San Giovanni, Italy. The technical performance (sensitivity and specificity and kappa coefficient) of the test performed on DBS was determined for different ratios (optical density/threshold value) compared to the results obtained on the plasma used as reference. We obtained a sensitivity of 100% with DBS for all ratios. The specificity increased according to the ratio of optical density of the individual EIA reaction to the threshold value, with 6.09%, 47.0%, and 83.0%, respectively, for ratios of 1.0, 3.0, and 5.0. Best performance was observed at ratio of 10.0 with a sensitivity and specificity of 100%. In conclusion, DBS eluate can be used for the diagnosis of viral hepatitis B and would be useful for conducting sero-epidemiological investigations. However, ratio giving best performance must be determined for each enzyme immunoassay method kits.
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Affiliation(s)
- Denis Rodrigue Kouamé
- Immunology Unit, UFR Pharmaceuticals and Biological Sciences, Félix Houphouët-Boigny University, BPV 34 Abidjan 01, Abidjan, Ivory Coast
| | - Roseline Affi Aboli
- Immunology Unit, UFR Pharmaceuticals and Biological Sciences, Félix Houphouët-Boigny University, BPV 34 Abidjan 01, Abidjan, Ivory Coast; Research and Diagnosis Center for AIDS and other infectious diseases (CeDReS), CHU (University Hospital) of Treichville, BP V3 Abidjan 01, Abidjan, Ivory Coast
| | - Mathieu Kabran
- Immunology Unit, UFR Pharmaceuticals and Biological Sciences, Félix Houphouët-Boigny University, BPV 34 Abidjan 01, Abidjan, Ivory Coast; Research and Diagnosis Center for AIDS and other infectious diseases (CeDReS), CHU (University Hospital) of Treichville, BP V3 Abidjan 01, Abidjan, Ivory Coast
| | - Aimé Cézaire Adiko
- Immunology Unit, UFR Pharmaceuticals and Biological Sciences, Félix Houphouët-Boigny University, BPV 34 Abidjan 01, Abidjan, Ivory Coast; Research and Diagnosis Center for AIDS and other infectious diseases (CeDReS), CHU (University Hospital) of Treichville, BP V3 Abidjan 01, Abidjan, Ivory Coast
| | - Mabarakissa Coulibaly
- Immunology Unit, UFR Pharmaceuticals and Biological Sciences, Félix Houphouët-Boigny University, BPV 34 Abidjan 01, Abidjan, Ivory Coast
| | - Bamory Dembele
- Immunology Unit, UFR Pharmaceuticals and Biological Sciences, Félix Houphouët-Boigny University, BPV 34 Abidjan 01, Abidjan, Ivory Coast; Laboratory of NBTS (National Blood Transfusion Center), BP V15 Abidjan 01, Ivory Coast.
| | - Andre Inwoley
- Immunology Unit, UFR Pharmaceuticals and Biological Sciences, Félix Houphouët-Boigny University, BPV 34 Abidjan 01, Abidjan, Ivory Coast; Research and Diagnosis Center for AIDS and other infectious diseases (CeDReS), CHU (University Hospital) of Treichville, BP V3 Abidjan 01, Abidjan, Ivory Coast
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9
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Miyano S, Pathammavong C, Ichimura Y, Sugiyama M, Phounphenghack K, Tengbriacheu C, Khamphaphongphane B, Nouanthong P, Franzel L, Yang TU, Raaijimakers H, Ota T, Funato M, Komada K, Hachiya M. Prevalence of hepatitis B and C virus infections in Lao People's Democratic Republic: The first national population-based cross-sectional survey. PLoS One 2022; 17:e0278933. [PMID: 36584043 PMCID: PMC9803141 DOI: 10.1371/journal.pone.0278933] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2022] [Accepted: 11/23/2022] [Indexed: 12/31/2022] Open
Abstract
Population-based seroprevalence of chronic hepatitis B and C infections has not been examined in Lao People's Democratic Republic (PDR). Therefore, this study aimed to estimate the seroprevalence of these infections in the general population of Lao PDR and perform subgroup analysis. A nationwide seroprevalence survey was conducted in Lao PDR in June 2019 using the multistage cluster sampling method. Dried blood spot samples were collected onto WhatmanTM 903 filter paper by finger prick. A chemiluminescent microparticle immunoassay was used to measure the levels of hepatitis B surface antigen (HBsAg) and hepatitis C antibody (HCV-Ab). Samples in which the HBsAg level was above 0.05 IU/ml and HCV-Ab was above the signal/cutoff ratio of 1.0 were considered positive based on comparisons with the relative light unit value of a calibration sample. A total of 1,927 samples (male: 47.3%, mean age: 23.0 years) were included in the analysis. The prevalence was estimated to be 4.2% (95% confidence interval [CI]: 2.7-6.3) for HBsAg and 1.6% (95% CI: 0.5-5.3) for HCV-Ab. Multivariable analysis revealed that those aged 20-24 years (adjusted odds ratio (AOR): 2.3, 95% CI: 1.1-4.6), those aged 25-29 years (AOR: 2.7, 95% CI: 1.3-5.6), those from the Northern region (AOR: 2.8, 95% CI: 1.2-6.6), and those who were Khmu (AOR: 3.6, 95% CI: 2.0-6.8) or Hmong (AOR: 5.0, 95% CI: 3.3-7.5) were significantly more likely to be positive for HBsAg. Although there were no statistically significant differences in the HCV-Ab prevalence according to each variable, males (2.9%, 95% CI: 0.7-10.7), those aged ≥40 years (6.1%, 95% CI: 2.1-16.8), and those from the Southern region (3.3%, 95% CI: 0.6-15.3) tended to have a higher prevalence. This novel population-based survey found differences in the prevalence of chronic hepatitis B and hepatitis C virus infections in Lao PDR according to sex, age group, region, and ethnicity; however, the results of this study should be confirmed in future studies, and relevant responses tailored for each target also need to be determined to control the transmission of hepatitis B and C infections.
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Affiliation(s)
- Shinsuke Miyano
- Bureau of International Health Cooperation and WHO Collaborating Center for Health Systems Development, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan
- * E-mail:
| | - Chansay Pathammavong
- National Immunization Program, Mother and Child Health Center, Ministry of Health, Lao People’s Democratic Republic (Lao PDR), Vientiane Capital, Lao PDR
| | - Yasunori Ichimura
- Bureau of International Health Cooperation and WHO Collaborating Center for Health Systems Development, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan
| | - Masaya Sugiyama
- Genome Medical Science Project, The Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Ichikwa, Chiba, Japan
| | - Kongxay Phounphenghack
- National Immunization Program, Mother and Child Health Center, Ministry of Health, Lao People’s Democratic Republic (Lao PDR), Vientiane Capital, Lao PDR
| | | | | | - Phonethipsavanh Nouanthong
- Institute Pasteur du Laos, National Immunization Technical Advisory Group, Ministry of Health, Vientiane Capital, Lao PDR
| | - Lauren Franzel
- Vaccine-Preventable Diseases and Immunization Team, WHO Lao PDR, Vientiane Capital, Lao PDR
| | - Tae Un Yang
- Vaccine-Preventable Diseases and Immunization Team, WHO Lao PDR, Vientiane Capital, Lao PDR
| | | | - Tomomi Ota
- Bureau of International Health Cooperation and WHO Collaborating Center for Health Systems Development, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan
| | - Masafumi Funato
- Bureau of International Health Cooperation and WHO Collaborating Center for Health Systems Development, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan
| | - Kenichi Komada
- Bureau of International Health Cooperation and WHO Collaborating Center for Health Systems Development, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan
| | - Masahiko Hachiya
- Bureau of International Health Cooperation and WHO Collaborating Center for Health Systems Development, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan
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10
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Okawa S, Komada K, Ichimura Y, Sugiyama M, Do HT, Le HX, Hoang TT, Nguyen TB, Huynh MK, Hoang HTH, Tran NAT, Le TH, Ngo QT, Miyano S, Yokobori Y, Inoue Y, Mizoue T, Hachiya M. Comparison between a rapid diagnostic test and dried blood spot-based immunoassay for hepatitis B surface antigen testing: Performance and cost implications in a population-based serosurvey in Vietnam. Int J Infect Dis 2022; 125:51-57. [PMID: 36241163 DOI: 10.1016/j.ijid.2022.10.011] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/08/2022] [Revised: 10/01/2022] [Accepted: 10/06/2022] [Indexed: 11/06/2022] Open
Abstract
OBJECTIVES This study aimed to determine the agreement between a rapid diagnostic test (RDT) and a dried blood spot (DBS)-based electrochemiluminescence immunoassay (ECLIA) of hepatitis B surface antigen and to compare the costs of conducting serosurveys using RDTs and DBS in a field setting. METHODS A serosurvey was conducted in the South Central Coast region of Vietnam in May 2019. Participants aged 1-39 years were recruited using a four-stage random sampling method and tested for hepatitis B surface antigen using an RDT kit (Alere Determine) and a DBS-based ECLIA. The agreement between the RDT and the DBS-based ECLIA was assessed using cross-tabulation and Cohen kappa. Cost data were categorized by input (personnel, transportation, field consumables, laboratory consumables, and capital item/overhead) and survey phase (survey preparation, data/biospecimen collection, laboratory testing, and coordination). RESULTS A total of 2072 participants were analyzed. There was a 99% agreement between the RDT and the DBS-based ECLIA results, with a Cohen kappa of 0.9. The estimated cost of conducting a serosurvey by DBS was UD $75,291, whereas RDT was $53,182. CONCLUSION RDTs and DBS-based ECLIA provide test results with high agreements. RDTs are a better option in terms of cost, whereas the DBS-based ECLIA may be useful when evaluating multiple infectious diseases.
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Affiliation(s)
- Sumiyo Okawa
- Bureau of International Health Cooperation, National Center for Global Health and Medicine, Sumiyo Okawa, 1-21-1, Toyama, Shinjuku-ku, Tokyo 162-8655, Japan.
| | - Kenichi Komada
- Bureau of International Health Cooperation, National Center for Global Health and Medicine, Sumiyo Okawa, 1-21-1, Toyama, Shinjuku-ku, Tokyo 162-8655, Japan
| | - Yasunori Ichimura
- Bureau of International Health Cooperation, National Center for Global Health and Medicine, Sumiyo Okawa, 1-21-1, Toyama, Shinjuku-ku, Tokyo 162-8655, Japan
| | - Masaya Sugiyama
- Genome Medical Science Project, The Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Chiba, Japan
| | - Hung Thai Do
- Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Huy Xuan Le
- Medical Health Service Center, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Thanh Tien Hoang
- Department of Infectious Disease Control and Prevention, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Trieu Bao Nguyen
- Department of Microbiology and Immunology, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Mai Kim Huynh
- Department of Microbiology and Immunology, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Hang Thi Hai Hoang
- Department of Infectious Disease Control and Prevention, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Nhu Anh Thi Tran
- Department of Microbiology and Immunology, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Thieu Hoang Le
- Department of Infectious Disease Control and Prevention, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Quyet Thi Ngo
- Department of Microbiology and Immunology, Pasteur Institute in Nha Trang, Nha Trang, Khánh Hòa, Viet Nam
| | - Shinsuke Miyano
- Bureau of International Health Cooperation, National Center for Global Health and Medicine, Sumiyo Okawa, 1-21-1, Toyama, Shinjuku-ku, Tokyo 162-8655, Japan
| | - Yuta Yokobori
- Bureau of International Health Cooperation, National Center for Global Health and Medicine, Sumiyo Okawa, 1-21-1, Toyama, Shinjuku-ku, Tokyo 162-8655, Japan
| | - Yosuke Inoue
- Department of Epidemiology and Prevention, National Center for Global Health and Medicine, Tokyo, Japan
| | - Tetsuya Mizoue
- Department of Epidemiology and Prevention, National Center for Global Health and Medicine, Tokyo, Japan
| | - Masahiko Hachiya
- Bureau of International Health Cooperation, National Center for Global Health and Medicine, Sumiyo Okawa, 1-21-1, Toyama, Shinjuku-ku, Tokyo 162-8655, Japan
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11
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Evaluation of blood samples collected by dried blood spots (DBS) method for hepatitis B virus DNA quantitation and its stability under real life conditions. JOURNAL OF CLINICAL VIROLOGY PLUS 2022. [DOI: 10.1016/j.jcvp.2022.100111] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/03/2022] Open
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12
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Kikuchi M, Lindstrom P, Tejada-Strop A, Mixson-Hayden T, Kamili S, Sawabe M. Dried blood spot is the feasible matrix for detection of some but not all hepatitis B virus markers of infection. BMC Res Notes 2022; 15:287. [PMID: 36064629 PMCID: PMC9446784 DOI: 10.1186/s13104-022-06178-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/11/2022] [Accepted: 08/25/2022] [Indexed: 11/30/2022] Open
Abstract
Objective Use of dried blood spots (DBS) for detection of hepatitis B virus (HBV) markers of infection has the potential to facilitate diagnosis of HBV infection especially in resource-limited countries. The aim of this study was to evaluate the feasibility of DBS for detection of various markers of HBV infections. Results Fifty-four DBS samples were engineered from well-characterized plasma samples. All DBS samples were tested for HBsAg, total anti-HBc and HBV DNA, 20 of 54 samples were also tested for HBeAg using commercially available assays. HBsAg was detected in 24 of 25 (96%), HBV DNA in 22 of 25 (88%), total anti-HBc in all 9 (100%), and HBeAg in all 7 (100%) DBS samples. The average difference in HBV DNA levels between DBS eluates and corresponding plasma samples was 2.7 log10 IU/mL. Fifteen DBS eluates positive for HBV DNA were sequenced and all of them belonged to HBV genotype A. Thirteen samples which were negative for all HBV markers showed HBeAg false positivity. Therefore, DBS is a reliable sample matrix for detection of HBsAg, total anti-HBc and HBV DNA, but not HBeAg. Further feasibility studies of DBS for diagnostic purposes and epidemiologic studies are warranted. Supplementary Information The online version contains supplementary material available at 10.1186/s13104-022-06178-x.
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Affiliation(s)
- Minami Kikuchi
- Division of Viral Hepatitis, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30329, USA.,Department of Molecular Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo, Tokyo, 113-8519, Japan
| | - Patrick Lindstrom
- Division of Viral Hepatitis, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30329, USA
| | - Alexandra Tejada-Strop
- Division of Viral Hepatitis, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30329, USA
| | - Tonya Mixson-Hayden
- Division of Viral Hepatitis, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30329, USA
| | - Saleem Kamili
- Division of Viral Hepatitis, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30329, USA
| | - Motoji Sawabe
- Department of Molecular Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo, Tokyo, 113-8519, Japan.
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13
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Shaver ZM, Anderson M, Bhebhe L, Baruti K, Chogaa WT, Ngidi J, Mbangiwa T, Taua M, Setlhare DR, Melamu P, Phinius BB, Musonda R, Mine M, Moyo S, Gaseitsiwe S. Decreased hepatitis B virus vaccine response among HIV-positive infants compared with HIV-negative infants in Botswana. AIDS 2022; 36:755-762. [PMID: 35113045 PMCID: PMC7614825 DOI: 10.1097/qad.0000000000003183] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
OBJECTIVES We sought to determine vaccine antibody titres and the prevalence of hepatitis B surface antigen (HBsAg) in both HIV-positive and HIV-negative infants born to HIV-positive mothers in Botswana. DESIGN This was a retrospective cross-sectional study using 449 archived dried blood spot samples from both HIV-positive and HIV-negative infants collected between 2016 and 2018. METHODS We screened dried blood spot samples for HBsAg and determined hepatitis B surface antibody titres. We determined hepatitis B virus (HBV) genotypes by amplifying 415 base-pairs of the surface region. RESULTS HIV-positive infants mounted a significantly lower immune response to the HBV vaccine (P < 0.001). Furthermore, a lower proportion of HIV-positive infants had protective hepatitis B surface antibody titres (74.5%) than HIV-negative infants (89.2%) (P < 0.001). HIV-positive infants were older and 50.9% of them had completed vaccination (P = 0.018). Of the 449 infant samples tested, three (0.67%) were positive for HBsAg. Of the three HBsAg-positive infants, two had protective titres (>10 mIU/ml). Two of the three HBV-positive infants were infected with genotype D3 and had no drug-resistance or escape mutations. CONCLUSION Vaccine response was lower among HIV-positive infants compared with HIV-negative infants. HBV infections were observed in both HIV-positive and HIV-negative infants in Botswana. Studies to investigate additional preventive strategies to reduce HBV mother-to-child transmission are recommended.
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Affiliation(s)
| | | | | | - Kabo Baruti
- Botswana Harvard AIDS Institute Partnership, Gaborone
- University of Botswana, Department of Biological Sciences, Gabarone, Botswana
| | - Wonderful T. Chogaa
- Botswana Harvard AIDS Institute Partnership, Gaborone
- Division of Human Genetics, Department of Pathology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa
| | - Julia Ngidi
- Botswana Harvard AIDS Institute Partnership, Gaborone
- National Health Laboratory, Ministry of Health and Wellness, Gaborone, Botswana
| | | | - Modiri Taua
- Botswana Harvard AIDS Institute Partnership, Gaborone
- National Health Laboratory, Ministry of Health and Wellness, Gaborone, Botswana
| | - Ditiro R. Setlhare
- Botswana Harvard AIDS Institute Partnership, Gaborone
- National Health Laboratory, Ministry of Health and Wellness, Gaborone, Botswana
| | - Pinkie Melamu
- Botswana Harvard AIDS Institute Partnership, Gaborone
| | | | | | - Madisa Mine
- Botswana Harvard AIDS Institute Partnership, Gaborone
- National Health Laboratory, Ministry of Health and Wellness, Gaborone, Botswana
| | - Sikhulile Moyo
- Botswana Harvard AIDS Institute Partnership, Gaborone
- Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, USA
| | - Simani Gaseitsiwe
- Botswana Harvard AIDS Institute Partnership, Gaborone
- Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, USA
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14
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Périères L, Diallo A, Marcellin F, Nishimwe ML, Ba EH, Coste M, Lo G, Halfon P, Touré Kane C, Maradan G, Carrieri P, Diouf A, Shimakawa Y, Sokhna C, Boyer S. Hepatitis B in Senegal: A Successful Infant Vaccination Program but Urgent Need to Scale Up Screening and Treatment (ANRS 12356 AmBASS survey). Hepatol Commun 2022; 6:1005-1015. [PMID: 34918868 PMCID: PMC9035578 DOI: 10.1002/hep4.1879] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/06/2021] [Revised: 11/18/2021] [Accepted: 11/24/2021] [Indexed: 12/17/2022] Open
Abstract
Senegal introduced the infant hepatitis B virus (HBV) vaccination in 2004 and recently committed to eliminating hepatitis B by 2030. Updated epidemiological data are needed to provide information on the progress being made and to develop new interventions. We estimated the prevalence of hepatitis B surface antigen (HBsAg) in children and adults living in rural Senegal and assessed hepatitis B treatment eligibility. A cross-sectional population-based serosurvey of HBsAg was conducted in 2018-2019 in a large sample (n = 3,118) of residents living in the Niakhar area (Fatick region, Senegal). Individuals positive for HBsAg subsequently underwent clinical and biological assessments. Data were weighted for age and sex and calibrated to be representative of the area's population. Among the 3,118 participants, 206 were HBsAg positive (prevalence, 6.9%; 95% confidence interval [CI], 5.6-8.1). Prevalence varied markedly according to age group in individuals aged 0-4, 5-14, 15-34, and ≥35 years as follows: 0.0% (95% CI, 0.00-0.01); 1.5% (95% CI, 0.0-2.3); 12.4% (95% CI, 9.1-15.6); and 8.8% (95% CI, 6.1-11.5), respectively. Of those subsequently assessed, 50.9% (95% CI, 41.8-60.0) had active HBV infection; 4 (2.9%; 95% CI, 0.9-9.4) were eligible for hepatitis B treatment. Conclusion: In this first population-based serosurvey targeting children and adults in rural Senegal, HBsAg prevalence was very low in the former, meeting the World Health Organization's (WHO) < 1% HBsAg 2020 target; however, it was high in young adults (15-34 years old) born before the HBV vaccine was introduced in 2004. To reach national and WHO hepatitis elimination goals, general population testing (particularly for adolescents and young adults), care, and treatment scale-up need to be implemented.
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Affiliation(s)
- Lauren Périères
- Vecteurs-Infections Tropicales et Méditerranéennes (VITROME)Campus Institut de Recherche pour le Développement (IRD)-Universite Cheikh Anta DiopDakarSenegal
| | - Aldiouma Diallo
- Vecteurs-Infections Tropicales et Méditerranéennes (VITROME)Campus Institut de Recherche pour le Développement (IRD)-Universite Cheikh Anta DiopDakarSenegal
| | - Fabienne Marcellin
- Institut National de la Santé et de la Recherche MédicaleIRDSciences Economiques and Sociales de la Santé and Traitement de l'Information MédicaleInstitut des Sciences de la Santé Publique - ISSPAMAix-Marseille UniversityMarseilleFrance
| | - Marie Libérée Nishimwe
- Institut National de la Santé et de la Recherche MédicaleIRDSciences Economiques and Sociales de la Santé and Traitement de l'Information MédicaleInstitut des Sciences de la Santé Publique - ISSPAMAix-Marseille UniversityMarseilleFrance
| | - El Hadji Ba
- Vecteurs-Infections Tropicales et Méditerranéennes (VITROME)Campus Institut de Recherche pour le Développement (IRD)-Universite Cheikh Anta DiopDakarSenegal
| | - Marion Coste
- Institut National de la Santé et de la Recherche MédicaleIRDSciences Economiques and Sociales de la Santé and Traitement de l'Information MédicaleInstitut des Sciences de la Santé Publique - ISSPAMAix-Marseille UniversityMarseilleFrance.,Centre National de la Recherche ScientifiqueÉcole des Hautes Études en Sciences SocialesCentrale MarseilleAix-Marseille School of EconomicsAix-Marseille UniversityMarseilleFrance
| | - Gora Lo
- Institut de Recherche en Santé de Surveillance Epidémiologique et de FormationDakarSenegal
| | | | - Coumba Touré Kane
- Institut de Recherche en Santé de Surveillance Epidémiologique et de FormationDakarSenegal
| | - Gwenaëlle Maradan
- Institut National de la Santé et de la Recherche MédicaleIRDSciences Economiques and Sociales de la Santé and Traitement de l'Information MédicaleInstitut des Sciences de la Santé Publique - ISSPAMAix-Marseille UniversityMarseilleFrance.,Observatoire Régional de la Santé Provence-Alpes-Côte d'AzurMarseilleFrance
| | - Patrizia Carrieri
- Institut National de la Santé et de la Recherche MédicaleIRDSciences Economiques and Sociales de la Santé and Traitement de l'Information MédicaleInstitut des Sciences de la Santé Publique - ISSPAMAix-Marseille UniversityMarseilleFrance
| | - Assane Diouf
- Vecteurs-Infections Tropicales et Méditerranéennes (VITROME)Campus Institut de Recherche pour le Développement (IRD)-Universite Cheikh Anta DiopDakarSenegal
| | - Yusuke Shimakawa
- Unité d'Épidémiologie des Maladies ÉmergentesInstitut PasteurParisFrance
| | - Cheikh Sokhna
- IRDService de santé des arméesVITROMEAix-Marseille UniversityMarseilleFrance
| | - Sylvie Boyer
- Institut National de la Santé et de la Recherche MédicaleIRDSciences Economiques and Sociales de la Santé and Traitement de l'Information MédicaleInstitut des Sciences de la Santé Publique - ISSPAMAix-Marseille UniversityMarseilleFrance
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15
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Bezerra CS, Portilho MM, Barbosa JR, de Azevedo CP, Mendonça ACDF, da Cruz JNM, Frota CC, do Lago BV, Villar LM. Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection. Sci Rep 2022; 12:1651. [PMID: 35102169 PMCID: PMC8803841 DOI: 10.1038/s41598-022-05264-1] [Citation(s) in RCA: 6] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/07/2021] [Accepted: 01/10/2022] [Indexed: 02/06/2023] Open
Abstract
Hepatitis B virus (HBV) diagnosis is performed on serum samples, but the access to this diagnosis is difficult in low-income regions. The use of dried blood spot (DBS) samples does not require special structure for collection, storage or transport. This study evaluates the use of DBS for detection, quantification and sequencing of HBV DNA using in-house techniques. Two study groups were included: 92 HBsAg + individuals and 49 negative controls. Serum and DBS samples were submitted to quantitative and qualitative in-house PCR for S/pol genes, sequencing and phylogenetic analyses. Total of 84 serum samples were successfully amplified. Of them, 63 paired DBS were also positive in qualitative PCR. Qualitative PCR in DBS presented a sensitivity of 75% and specificity of 100% (Kappa = 0.689). Quantitative PCR in DBS presented a detection limit of 852.5 copies/mL (250 IU/mL), sensitivity of 77.63% and specificity of 100% (Kappa = 0.731). A total of 63 serum samples and 36 DBS samples were submitted to sequencing, revealing the circulation of genotypes A (65.08%), D (4.8%), E (3.2%) and F (27%) with 100% of correspondence between serum and DBS. All sequenced samples displayed polymorphisms in HBsAg gene. An HIV-coinfected patient presented the rtM204V/I-rtL180M double resistance mutation in serum and DBS. In conclusion, DBS is an alternative to detect, quantify and characterize HBV DNA, being a possibility of increasing diagnosis in low-income settings, closing gaps in HBV control.
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16
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Périères L, Protopopescu C, Lo G, Marcellin F, Ba EH, Coste M, Touré Kane C, Diallo A, Sokhna C, Boyer S. Sibling status, home birth, tattoos and stitches are risk factors for chronic hepatitis B virus infection in Senegalese children: A cross-sectional survey. J Viral Hepat 2021; 28:1515-1525. [PMID: 34355470 DOI: 10.1111/jvh.13589] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/05/2021] [Revised: 07/12/2021] [Accepted: 07/22/2021] [Indexed: 12/14/2022]
Abstract
Sub-Saharan Africa's hepatitis B virus (HBV) burden is primarily due to infection in infancy. However, data on chronic HBV infection prevalence and associated risk factors in children born post-HBV vaccination introduction are scarce. We estimated hepatitis B surface antigen (HBsAg) prevalence and risk factors in Senegalese children born during the HBV vaccination era. In 2018-2019, a community-based cross-sectional survey was conducted in Senegal among children born between 2004 and 2015 (ie after the three-dose HBV vaccine series was introduced (2004) but before the birth dose's introduction (2016)). HBsAg-positive children were identified using dried blood spots. A standardized questionnaire collected socioeconomic information. Data were age-sex weighted and calibrated to be representative of children living in the study area. Risk factors associated with HBsAg positivity were identified using negative binomial regression. Among 1,327 children, 17 were HBsAg-positive (prevalence = 1.23% (95% confidence interval [CI] 0.61-1.85)). Older age (adjusted incidence-rate ratio [aIRR] 1.31 per one-year increase, 95% CI 1.10-1.57), home vs healthcare facility delivery (aIRR 3.55, 95% CI 1.39-9.02), stitches (lifetime) (aIRR 4.79; 95% CI 1.84-12.39), tattoos (aIRR 8.97, 95% CI 1.01-79.11) and having an HBsAg-positive sibling with the same mother (aIRR 3.05, 95% CI 1.09-8.57) were all independently associated with HBsAg positivity. The low HBsAg prevalence highlights the success of the Senegalese HBV vaccination program. To further reduce HBV acquisition in children, high-risk groups, including pregnant women and siblings of HBsAg-positive individuals, must be screened. Vital HBV infection prevention measures include promoting delivery in healthcare facilities, and increasing awareness of prevention and control procedures.
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Affiliation(s)
| | - Camelia Protopopescu
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Économiques & Sociales de la Santé & Traitement de l'Information Médicale, ISSPAM, Marseille, France
| | - Gora Lo
- Institut de Recherche en Santé de Surveillance Epidémiologique et de Formation, Dakar, Senegal
| | - Fabienne Marcellin
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Économiques & Sociales de la Santé & Traitement de l'Information Médicale, ISSPAM, Marseille, France
| | | | - Marion Coste
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Économiques & Sociales de la Santé & Traitement de l'Information Médicale, ISSPAM, Marseille, France.,Aix Marseille Univ, CNRS, EHESS, Centrale Marseille, AMSE, Marseille, France
| | - Coumba Touré Kane
- Institut de Recherche en Santé de Surveillance Epidémiologique et de Formation, Dakar, Senegal
| | | | - Cheikh Sokhna
- VITROME, IRD, AMU, AP-HM, SSA, IHU-MI, Marseille, France
| | - Sylvie Boyer
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Économiques & Sociales de la Santé & Traitement de l'Information Médicale, ISSPAM, Marseille, France
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17
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Jackson K, Tekoaua R, Li X, Locarnini S. Real-world application of the Xpert® HBV viral load assay on serum and dried blood spots. J Med Virol 2021; 93:3707-3713. [PMID: 33174623 DOI: 10.1002/jmv.26662] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/23/2020] [Revised: 11/03/2020] [Accepted: 11/08/2020] [Indexed: 12/31/2022]
Abstract
As we strive towards the WHO goal of elimination of viral hepatitis as a public health threat by 2030, implementation of reliable, accurate diagnostic assays is crucial to identify those at risk of disease progression and those at risk of transmission. Ironically those at greatest risk of chronic hepatitis B are often in resource-poor regions with limited access to testing, collection, storage, and/or transportation of peripheral blood. The Xpert® HBV Viral Load assay provides an easy to use, convenient means of measuring load on GeneXpert platforms. In this study, the Xpert assay is evaluated against four commercially available high-throughput assays for Hepatitis B virus (HBV) loads. In addition application of dried blood spots (DBS) for estimation of viral load is assessed on real-world samples collected from a remote Pacific Island, Kiribati. A total of 107 serum/plasma samples were tested in the Xpert HBV load assay and compared with the Abbott m2000, Alinity m, and Roche Cobas CAP/CTM and 6800. Fifty-three DBS were tested in the Xpert assay and compared with matching serum samples. Overall 82% serum/plasma samples demonstrated good correlation between the Xpert and Roche and Abbott assays, to within 0.5 log10 IU/ml. The greatest discrepancies were seen at the limits of quantification of all assays. About 85.4% DBS gave estimable viral loads to within 1 log10 IU/ml of the serum load. The Xpert HBV viral load assay is recommended for all settings but particularly useful for resource-poor settings. Utility of DBS with the Xpert assay provides a simple means for testing in remote settings.
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Affiliation(s)
- Kathy Jackson
- Victorian Infectious Diseases Reference Laboratory, Melbourne Health, Doherty Institute for Infection and Immunity, Melbourne, Australia
| | - Rosemary Tekoaua
- Ministry of Health and Medical Services, Tungaru Central Hospital, Tarawa, Republic of Kiribati
| | - Xin Li
- Victorian Infectious Diseases Reference Laboratory, Melbourne Health, Doherty Institute for Infection and Immunity, Melbourne, Australia
| | - Stephen Locarnini
- Victorian Infectious Diseases Reference Laboratory, Melbourne Health, Doherty Institute for Infection and Immunity, Melbourne, Australia
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18
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Bezerra CS, Portilho MM, Frota CC, Villar LM. Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples. Microbiologyopen 2021; 10:e1161. [PMID: 33970537 PMCID: PMC8107022 DOI: 10.1002/mbo3.1161] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/15/2020] [Revised: 12/22/2020] [Accepted: 12/29/2020] [Indexed: 01/15/2023] Open
Abstract
The dried blood spot (DBS) samples are a useful resource for viral DNA isolation and important in increasing access to HBV diagnosis. However, the choice of the DNA extraction method is crucial for reliable results. We compared the reliability of four DNA extraction methods using DBS samples for the qualitative and quantitative detection of HBV. A panel of serially diluted HBV DNA in whole blood was spotted onto filter paper (Whatman 903 paper and Whatman FTA cards). Four methods were used to extract DNA: QIAamp® DNA Blood Mini Kit (Qiagen); High Pure Viral Nucleic Acid Kit (Roche); Invisorb Spin Blood Midi Kit (Invitek), and DBS Genomic DNA Isolation Kit (Norgen Biotek). Two qualitative PCRs for the core and surface gene regions of HBV were used, and in‐house real‐time PCR was also evaluated. It was possible to detect HBV DNA using all extraction and PCR protocols. The lowest limit of detection was found using Whatman 903 paper, Roche extraction, and qualitative PCR (20 copies of HBV DNA per ml) for the surface/polymerase HBV region. In the case of in‐house real‐time PCR, the lowest limit of detection was found using both Roche and Qiagen assays (estimated 2 × 103 copies per ml). These results suggest the importance of both the extraction method and PCR protocol in detecting HBV DNA in DBS. This study provides insights into the utility of DBS samples in HBV molecular diagnosis and their feasibility in low resource areas where cold storage and transportation may be difficult.
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Affiliation(s)
- Cristianne Sousa Bezerra
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Fundação Oswaldo Cruz (Fiocruz, Rio de Janeiro, Brazil.,Departamento de Educação, Instituto Federal de Educação, Ciência e Tecnologia do Ceará, Fortaleza, Brazil
| | - Moyra Machado Portilho
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Fundação Oswaldo Cruz (Fiocruz, Rio de Janeiro, Brazil.,Instituto de Pesquisas Gonçalo Moniz, Fiocruz, Salvador, Brazil
| | - Cristiane Cunha Frota
- Departamento de Patologia e Medicina Legal, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, Brazil
| | - Lívia Melo Villar
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Fundação Oswaldo Cruz (Fiocruz, Rio de Janeiro, Brazil
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19
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Amini F, Auma E, Hsia Y, Bilton S, Hall T, Ramkhelawon L, Heath PT, Le Doare K. Reliability of dried blood spot (DBS) cards in antibody measurement: A systematic review. PLoS One 2021; 16:e0248218. [PMID: 33720928 PMCID: PMC7959368 DOI: 10.1371/journal.pone.0248218] [Citation(s) in RCA: 38] [Impact Index Per Article: 9.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2020] [Accepted: 02/23/2021] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Increasingly, vaccine efficacy studies are being recommended in low-and-middle-income countries (LMIC), yet often facilities are unavailable to take and store infant blood samples correctly. Dried blood spots (DBS), are useful for collecting blood from infants for diagnostic purposes, especially in low-income settings, as the amount of blood required is miniscule and no refrigeration is required. Little is known about their utility for antibody studies in children. This systematic review aims to investigate the correlation of antibody concentrations against infectious diseases in DBS in comparison to serum or plasma samples that might inform their use in vaccine clinical trials. METHODS AND FINDINGS We searched MEDLINE, Embase and the Cochrane library for relevant studies between January 1990 to October 2020 with no language restriction, using PRISMA guidelines, investigating the correlation between antibody concentrations in DBS and serum or plasma samples, and the effect of storage temperature on DBS diagnostic performance. We included 40 studies in this systematic review. The antibody concentration in DBS and serum/plasma samples reported a good pooled correlation, (r2 = 0.86 (ranged 0.43 to 1.00)). Ten studies described a decline of antibody after 28 days at room temperature compared to optimal storage at -20°C, where antibodies were stable for up to 200 days. There were only five studies of anti-bacterial antibodies. CONCLUSIONS There is a good correlation between antibody concentrations in DBS and serum/plasma samples, supporting the wider use of DBS in vaccine and sero-epidemiological studies, but there is limited data on anti-bacterial antibodies. The correct storage of DBS is critical and may be a consideration for longer term storage.
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Affiliation(s)
- Fahimah Amini
- Paediatric Infectious Disease Research Group, Institute for Infection and Immunity, St. George’s University of London, London, United Kingdom
| | - Erick Auma
- Department of Biology, University of Lyon, Université Claude Bernard Lyon, ENS de Lyon, CNRS, UMR, Lyon, France
| | - Yingfen Hsia
- Paediatric Infectious Disease Research Group, Institute for Infection and Immunity, St. George’s University of London, London, United Kingdom
- School of Pharmacy, Queen’s University Belfast, Belfast, United Kingdom
| | - Sam Bilton
- St Georges University Hospitals NHS Foundation Trust, Tooting, London, United Kingdom
| | - Tom Hall
- Paediatric Infectious Disease Research Group, Institute for Infection and Immunity, St. George’s University of London, London, United Kingdom
| | - Laxmee Ramkhelawon
- Paediatric Infectious Disease Research Group, Institute for Infection and Immunity, St. George’s University of London, London, United Kingdom
| | - Paul T. Heath
- Paediatric Infectious Disease Research Group, Institute for Infection and Immunity, St. George’s University of London, London, United Kingdom
- St Georges University Hospitals NHS Foundation Trust, Tooting, London, United Kingdom
| | - Kirsty Le Doare
- Paediatric Infectious Disease Research Group, Institute for Infection and Immunity, St. George’s University of London, London, United Kingdom
- St Georges University Hospitals NHS Foundation Trust, Tooting, London, United Kingdom
- Pathogen Immunology Group, Public Health England, Porton Down, United Kingdom
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20
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Martínez-Campreciós J, Rando-Segura A, Buti M, Rodrigo-Velásquez F, Riveiro-Barciela M, Barreira-Díaz A, Álvarez-López P, Salmerón P, Palom A, Tabernero D, Palomo N, Nindia A, Barbosa G, López E, Ferreira V, Saiago N, Kuchta A, Ferrer-Costa R, Esteban R, Molina I, Rodríguez-Frías F. Reflex viral load testing in dried blood spots generated by plasma separation card allows the screening and diagnosis of chronic viral hepatitis. J Virol Methods 2021; 289:114039. [PMID: 33338545 DOI: 10.1016/j.jviromet.2020.114039] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/21/2020] [Revised: 12/03/2020] [Accepted: 12/06/2020] [Indexed: 02/07/2023]
Abstract
Dried blood spots (DBS) have been proposed as an alternative diagnostic technique for chronic viral hepatitis. The aim of this observational study was to correlate serologic HBV, HCV, and HDV status and reflex the respective viral load testing by PSC-DBS samples from capillary blood vs conventional plasma samples in patients with chronic viral hepatitis. Besides, we apply these tests in a prospective study for chronic viral hepatitis diagnosis in a rural region of sub-Saharan Africa. In total, 124 HBsAg-positive patients, 75 anti-HCV positive, 2 with HBV-HCV coinfection, and 13 anti-HDV positive were included. PSC-DBS sensitivity/specificity was 98.4 %/96.2 % for HBsAg detection, 98.7 %/100 % for anti-HCV, and 84.6 %/100 % for anti-HDV. HCV-RNA was quantified in all viremic patients using DBS. Only 42 of 78 (53.8 %) samples with HBV-DNA viremia were quantifiable by DBS. Sensitivity increased to 95.7 % in patients with HBV-DNA levels >2000 IU/mL. There was a high correlation between DBS and venous blood. The prevalence of HBsAg among the 93 individuals tested in Angola was 11 %, and 60 % of cases had detectable HBV-DNA viremia. As a conclusion, PSC-DBS is useful for chronic viral hepatitis screening and reflex molecular diagnosis showing globally high sensitivities and correlation with conventional blood samples.
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Affiliation(s)
- Joan Martínez-Campreciós
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain; Universitat Autònoma de Barcelona, 08193, Bellaterra, Spain
| | - Ariadna Rando-Segura
- Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain; Department of Microbiology, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Universitat Autònoma de Barcelona, 08193, Bellaterra, Spain
| | - María Buti
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain; CIBERehd, Instituto Carlos II, Spain.
| | - Fernando Rodrigo-Velásquez
- Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain; Department of Microbiology, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Universitat Autònoma de Barcelona, 08193, Bellaterra, Spain
| | - Mar Riveiro-Barciela
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain; CIBERehd, Instituto Carlos II, Spain
| | - Ana Barreira-Díaz
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain
| | - Patricia Álvarez-López
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain
| | - Paula Salmerón
- Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain; Department of Microbiology, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Universitat Autònoma de Barcelona, 08193, Bellaterra, Spain
| | - Adriana Palom
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain
| | - David Tabernero
- CIBERehd, Instituto Carlos II, Spain; Liver Pathology Unit, Biochemistry and Microbiology Departments, Hospital Universitari Vall d'Hebron, Barcelona, Spain
| | - Nieves Palomo
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain
| | | | | | - Eva López
- Hospital Nossa Senhora da Paz, Cubal, Angola
| | - Vicelma Ferreira
- Hospital General de Benguela, Universidade Katyavla Bwila, Benguela, Angola
| | - Nelsa Saiago
- Hospital General de Benguela, Universidade Katyavla Bwila, Benguela, Angola
| | | | - Roser Ferrer-Costa
- Biochemistry Department, Clinical Laboratories Hospital Universitari Vall d'Hebron, Spain
| | - Rafael Esteban
- Liver Unit, Internal Medicine Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain; Vall d'Hebron Barcelona Hospital Campus, Passeig Vall d'Hebron 119-129, 080335 Barcelona, Spain; CIBERehd, Instituto Carlos II, Spain
| | - Israel Molina
- Infectious Diseases Department, Hospital Universitari Vall d'Hebron, PROSICS Barcelona, Universitat Autònoma de Barcelona, Barcelona, Spain
| | - Francisco Rodríguez-Frías
- CIBERehd, Instituto Carlos II, Spain; Biochemistry Department, Clinical Laboratories Hospital Universitari Vall d'Hebron, Spain; Liver Pathology Unit, Biochemistry and Microbiology Departments, Hospital Universitari Vall d'Hebron, Barcelona, Spain
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21
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Wannaratana S, Thontiravong A, Pakpinyo S. Comparison of three filter paper -based devices for safety and stability of viral sample collection in poultry. Avian Pathol 2020; 50:78-84. [PMID: 33059461 DOI: 10.1080/03079457.2020.1837343] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
General diagnosis of poultry viruses primarily relies on detection of viruses in samples, but many farms are located in remote areas requiring logistic transportation. Filter paper cards are a useful technology that offer an alternative for collecting and preserving samples without hazardous exposure. The goal of this study was to compare three filter papers: the Flinders Technology Associates filter (FTA®) card, dried blood spot (DBS) card and qualitative filter paper (FP) grade 2 to collect poultry samples. In particular, we have used Newcastle disease virus (NDV) to evaluate safety and a Marek's disease virus (MDV) attenuated vaccine (CVI988) to evaluate stability of viral DNA. This experiment was divided into two parts. The first part was to determine the DNA stability and detection limit of CVI988 in samples collected in different paper supports after four storage times (3, 7, 14 and 30 days post spot). The second part was to determine the safety of papers by evaluating the viral inactivation efficacy using NDV as a representative virus. Results showed that all papers could preserve CVI988 DNA at all times, with a detection limit of 0.5 PFU/5 µl for FTA® and DBS cards, and 5 PFU/5 µl for FP. Our results showed that the NDV remained viable and infectious on the DBS card and FP, while no viable virus was detected on the FTA® card, suggesting that the FTA® card was safest to use. Therefore, the use of the DBS card and FP for infectious sample collection should be discouraged and reconsidered. RESEARCH HIGHLIGHTS The detection limits of the FTA® card, DBS card and FP for CVI988 detection were 0.5, 0.5 and 5 PFU/5 µl, respectively. All three filter papers could preserve viral DNA for at least 30 days of post spot. The DBS card and FP are not suitable for collecting NDV samples, which is one of the major economical threats for the poultry industry worldwide.
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Affiliation(s)
- Suwarak Wannaratana
- Faculty of Veterinary Medicine, Rajamangala University of Technology Tawan-Ok, Chonburi, Thailand
| | - Aunyaratana Thontiravong
- Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.,Center of Excellence for Emerging and Re-emerging Infectious Diseases in Animals (CUEIDAs), Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand
| | - Somsak Pakpinyo
- Center of Excellence for Emerging and Re-emerging Infectious Diseases in Animals (CUEIDAs), Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.,Avian Health Research Unit, Department of Veterinary Medicine, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand
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22
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Evaluation of dried blood spot testing using the Abbott Alinity i. J Clin Virol 2020; 132:104638. [PMID: 33049642 DOI: 10.1016/j.jcv.2020.104638] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/17/2020] [Revised: 09/03/2020] [Accepted: 09/06/2020] [Indexed: 11/23/2022]
Abstract
BACKGROUND The West of Scotland Specialist Virology Centre currently uses the Abbott Architect for DBS serology. The new Abbott Alinity i will replace the Architect in our laboratory. In this study, mock and stored patient DBS samples were tested on both platforms and results compared. STUDY DESIGN Mock DBS were made from whole blood where patient results were known (38 negative samples and 141 positive samples; 39 HIV Antigen/Antibody (Ag/Ab), 35 HCV IgG antibody (HCVG), 34 HBV core IgG (HBCG) and 33 HBsAg). Mock DBS were tested on both Abbott platforms. Stored patient DBS samples (132 negative and 263 positive: 9 HIVAg/Ab, 10 HBsAg, 52 HBCG and 60 HCVG) previously tested on the Architect were retested on the Alinity i. RESULTS Mock DBS showed good correlation between the Architect and Alinity i for the HIV Ag/Ab,HBCG and HCVG assays. A poorer correlation occurred with HBsAg, the Alinity i reported HBsAg positives at a lower value compared to the Architect. The coefficient of variation for intra-assay variation was 1.69 % (HIVAg/Ab), 3.25 % (HCVG), 1.68 % (HBsAg) and 1.95 % (HBCG). The sensitivity and specificity was determined based on results from the mock and patient samples. At S/Co cut-off 1.0 both HIV and HBsAg had a sensitivity of 100 %. A cut-off 0.8 gave a sensitivity of 95.83 % (95 % CI 89.67%-98.85%) for HCVG and 0.3 gave a sensitivity of 98.8 % (CI 93.69%-99.97%) for HBCG. DISCUSSION The alinity i compared well against the architect and can be used to test DBS samples.
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23
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Roger S, Lefeuvre C, Grison M, Ducancelle A, Lunel-Fabiani F, Pivert A, Le Guillou-Guillemette H. Evaluation of the Aptima™ HBV Quant Dx assay for semi-quantitative HBV viral load from dried blood spots. J Clin Virol 2020; 129:104524. [PMID: 32629186 DOI: 10.1016/j.jcv.2020.104524] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2020] [Revised: 06/17/2020] [Accepted: 06/22/2020] [Indexed: 01/15/2023]
Abstract
BACKGROUND The detection and quantification of hepatitis B virus (HBV) DNA from dried blood spots (DBS) is a major tool for chronic hepatitis B management in resource-limited settings. This strategy fits in perfectly with the hepatitis control plan promoted by the World Health Organization. However, few commercial methods are validated for viral load (VL) measurement on DBS. OBJECTIVES Our objective was to evaluate the performance of the HBV VL measurement of the Aptima™ HBV Quant Dx assay on DBS compared to plasma samples on the Panther® platform (Hologic). STUDY DESIGN 266 whole blood samples for routine measurement were included. Five spots of 75 μL of whole blood were loaded onto a card before centrifugation and plasma settling. RESULTS 149 samples were quantifiable and 117 were not detected. We achieved excellent linearity (r² = 0.994) over a wide range of measurements suitable for clinical practice, and a 95 % lower limit of detection (LLOD-95 %) at 2.65 log10 IU/mL (445 IU/mL). A good performance of this assay was observed for samples with HBV VL > LLOD-95 % and 100 % of samples were detected if HBV VL was above 2.95 log10 IU/mL. The correlation between the two matrices for quantitative VLs was good (r² = 0.978) with a very low bias (-0.002 log10 IU/mL). CONCLUSION The Aptima™ assay can properly detect and quantify HBV DNA in DBS, providing a satisfactory use in clinical monitoring and therapeutic decisions. DBS represents an excellent alternative to plasma, especially in resource-limited countries, while maintaining the performance and advantages of an automated technique.
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Affiliation(s)
- Steven Roger
- Virology Department, Angers University Hospital, Angers, France
| | - Caroline Lefeuvre
- Virology Department, Angers University Hospital, Angers, France; HIFIH Laboratory EA 3859, LUNAM, Angers, France
| | - Marine Grison
- Virology Department, Angers University Hospital, Angers, France
| | - Alexandra Ducancelle
- Virology Department, Angers University Hospital, Angers, France; HIFIH Laboratory EA 3859, LUNAM, Angers, France
| | - Françoise Lunel-Fabiani
- Virology Department, Angers University Hospital, Angers, France; HIFIH Laboratory EA 3859, LUNAM, Angers, France
| | - Adeline Pivert
- Virology Department, Angers University Hospital, Angers, France; HIFIH Laboratory EA 3859, LUNAM, Angers, France
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24
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Poiteau L, Wlassow M, Hézode C, Pawlotsky JM, Chevaliez S. Evaluation of the Xpert HBV Viral Load for hepatitis B virus molecular testing. J Clin Virol 2020; 129:104481. [PMID: 32512377 DOI: 10.1016/j.jcv.2020.104481] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/04/2020] [Revised: 05/23/2020] [Accepted: 05/31/2020] [Indexed: 12/26/2022]
Abstract
BACKGROUND Accurate molecular methods to detect and quantify hepatitis B virus (HBV) DNA are essential to diagnose chronic infections, guide treatment decisions, assess response to treatment, and determine risk of HBV-related complications. New HBV DNA generations of real-time assay platforms are now available with the availability of results in less than 2-3 h and a continuous loading of specimens with true random access. OBJECTIVES The aim of this prospective study was to evaluate the performance of the new Xpert HBV Viral Load assay to accurately quantify HBV DNA in plasma and in whole blood collected on dried blood spot (DBS). METHODS Plasma and whole blood from 143 patients chronically infected with HBV were tested in parallel using two commercially real-time PCR assay (Cobas AmpliPrep/Cobas Taqman HBV test, version 2.0, and Xpert HBV Viral Load assay). RESULTS HBV DNA levels in whole blood strongly correlated with those measured in plasma. A positive significant correlation between the HBV DNA levels in plasma measured with the new Xpert HBV Viral Load and CAP/CTM HBV v2.0 assays was found. CONCLUSIONS The newly developed real-time PCR-based assay Xpert HBV Viral Load accurately quantifies HBV DNA in whole blood specimens as well as in plasma samples from patients with chronic HBV infection. Whole blood specimens collected on DBS can be confidently used for replicative HBV infection detection in patients with HBV DNA level above 3 Log using standardized, commercially available methods.
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Affiliation(s)
- Lila Poiteau
- National Reference Center for Viral Hepatitis B, C and Delta, Department of Virology, Hôpital Henri Mondor, France; INSERM U955, Créteil, France
| | - Mélanie Wlassow
- National Reference Center for Viral Hepatitis B, C and Delta, Department of Virology, Hôpital Henri Mondor, France; INSERM U955, Créteil, France
| | - Christophe Hézode
- Department of Hepatology and Gastroenterology, Hôpital Henri Mondor, France; INSERM U955, Créteil, France
| | - Jean-Michel Pawlotsky
- National Reference Center for Viral Hepatitis B, C and Delta, Department of Virology, Hôpital Henri Mondor, France; INSERM U955, Créteil, France
| | - Stéphane Chevaliez
- National Reference Center for Viral Hepatitis B, C and Delta, Department of Virology, Hôpital Henri Mondor, France; INSERM U955, Créteil, France.
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Abstract
Human immunodeficiency virus (HIV), a type of lentivirus (a subgroup of retrovirus), causes acquired immunodeficiency syndrome (AIDS). This pathophysiologic state destroys the immune system allowing opportunistic infections, cancer and other life-threatening diseases to thrive. Although many analytic tools including enzyme-linked immunoassay (ELISA), indirect and line immunoassay, Western blotting, radio-immunoprecipitation, nucleic acid amplification testing (NAAT) have been developed to detect HIV, recent developments in nanosensor technology have prompted its use as a novel diagnostic approach. Nanosensors provide analytical information about behavior and characteristics of particles by using biochemical reactions mediated by enzymes, immune components, cells and tissues. These reactions are transformed into decipherable signals, i.e., electrical, thermal, optical, using nano to micro scale technology. Nanosensors are capable of both quantitative and qualitative detection of HIV, are highly specific and sensitive and provide rapid reproducible results. Nanosensor technology can trace infant infection during mother-to-child transmission, the latent HIV pool and monitor anti-HIV therapy. In this chapter, we review nanosensor analytics including electrochemical, optical, piezoelectric, SERS-based lateral flow assay, microfluidic channel-based biosensors in the detection of HIV. Other techniques in combination with different biorecognition elements (aptamers, antibodies, oligonucleotides) are also discussed.
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Affiliation(s)
- Sarthak Nandi
- DBT-National Institute of Animal Biotechnology (DBT-NIAB), Hyderabad, Telangana, India
| | - Ayusi Mondal
- DBT-National Institute of Animal Biotechnology (DBT-NIAB), Hyderabad, Telangana, India
| | - Akanksha Roberts
- DBT-National Institute of Animal Biotechnology (DBT-NIAB), Hyderabad, Telangana, India
| | - Sonu Gandhi
- DBT-National Institute of Animal Biotechnology (DBT-NIAB), Hyderabad, Telangana, India.
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Evaluation of the efficiency of dried blood spot-based measurement of hepatitis B and hepatitis C virus seromarkers. Sci Rep 2020; 10:3857. [PMID: 32123234 PMCID: PMC7052143 DOI: 10.1038/s41598-020-60703-1] [Citation(s) in RCA: 22] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/07/2019] [Accepted: 02/10/2020] [Indexed: 01/26/2023] Open
Abstract
Although hepatitis B (HBV) and C (HCV) virus infections are still global health issues, measuring sero-markers by standard venipuncture is challenging in areas limited with the adequate human resources and basic infrastructure. This study aimed to inform the usefulness of dried blood spot (DBS) sampling technique for epidemiological study of HBV and HCV in the resources limited areas. We compared specimen recovery rate expressed as analytical sensitivity ratio of HBsAg, HBcAb and anti-HCV between serum specimens and DBS samples (HemaSpot vs Whatman903). Sensitivity ratio was calculated as the ratio of the measured value from DBS to the measured value from serum. Then both the qualitative and quantitative comparisons of HBsAg detection by DBS were done using Cambodian samples. HBsAg, HBcAb and anti-HCV sensitivity ratios for the highest sample dilution (8-fold) were 31.2:1, 38.9:1 and 32.0:1 for Whatman903 card and 17.6:1, 23.5:1 and 26.3:1 for HemaSpot respectively. Detection efficacy of HemaSpot (80%) was not inferior to Whatman903 (60%) after 1 month storage, and no significant difference in any hepatitis virus sero-markers was observed in HemaSpot-spotted patient samples stored for 2 weeks at −25 °C and 29 °C. All reference HemaSpot -spotted 400 HBsAg sero-negative samples showed negative. Sensitivity and specificity of HBsAg in HemaSpot were 92.3% and 100%. The recovery expressed as analytical sensitivity ratio of HBsAg, HBcAb and anti-HCV of HemaSpot specimen were not inferior to Whatman903. Therefore, DBS with its usefulness proved as an acceptable tool for large epidemiological study of HBV and HCV in resources limited remote area.
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Cruz HM, de Paula VS, Cruz JCM, do Ó KMR, Milagres FAP, Bastos FI, Mota JCD, Cruz MS, Andrade TMD, Pollo-Flores P, Leal E, Motta-Castro ARC, Ivantes CAP, Bezerra CS, Barbosa JR, da Cruz JNM, Lewis-Ximenez LL, Villar LM. Evaluation of accuracy of hepatitis B virus antigen and antibody detection and relationship between epidemiological factors using dried blood spot. J Virol Methods 2019; 277:113798. [PMID: 31837375 DOI: 10.1016/j.jviromet.2019.113798] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2019] [Revised: 10/15/2019] [Accepted: 12/10/2019] [Indexed: 02/07/2023]
Abstract
Dried blood spots (DBS) testing might increase the access for Hepatitis B virus (HBV) diagnosis, but little is known about the performance of these assays in real life conditions. This study aims to evaluate the diagnostic accuracy of HBsAg, anti-HBc and anti-HBs detection in DBS in clinical settings and field studies and to evaluate demographic and risk behaviour according the presence of HBsAg and anti-HBc. Paired sera and DBS samples were obtained from 2309 individuals from 3 groups, defined as follows: G1: clinical setting (n = 5-19), G2: general population (n = 1305) and G3: vulnerable individuals that could be more exposed to blood contact (n = 485). Sera and DBS were tested using commercial enzyme immunoassay (EIA), with some modifications added. Using DBS samples, the specificity values were above 90 % for HBsAg and anti-HBc in all groups and for anti-HBs range from 58.6%-85%. HBsAg testing had the best performance in GI (sensitivity = 84.4 %) and among those samples that the paired serum also presented anti-HBc marker (sensitivity = 91.6 %). High sensitivity of anti-HBc testing in DBS samples was observed in GI (80.8 %) and among HBV active cases (HBsAg+/anti-HBc+) (98.4 %). Testing of anti-HBs in DBS showed the highest sensitivity in GIII (65.5 %), in previous HBV exposed and cured individuals and when serum titers were above 100 IU/mL (86.7 %). DBS samples could be used for screening and prevalence studies for HBsAg and anti-HBc, particularly in clinical settings and among HBV active cases in field studies.
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Affiliation(s)
- Helena Medina Cruz
- Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil
| | | | | | | | | | - Francisco Inácio Bastos
- Institute of Communication and Scientific Information & Technology for Health, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil
| | - Jurema Corrêa da Mota
- Institute of Communication and Scientific Information & Technology for Health, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil
| | - Marcelo Santos Cruz
- Institute of Psychiatry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil
| | | | - Priscila Pollo-Flores
- Antonio Pedro University Hospital, Federal Fluminense University, Rio de Janeiro, Brazil
| | - Erotildes Leal
- Federal University of Rio de Janeiro, Faculty of Medicine, Rio de Janeiro, Brazil
| | | | | | - Cristianne Sousa Bezerra
- Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil; Instituto Federal de Ciencia e Tecnologia do Ceará, Brazil
| | - Jakeline Ribeiro Barbosa
- Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil; FIOCRUZ, Brasilia, Brazil
| | | | | | - Livia Melo Villar
- Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil.
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Villar LM, Cruz HM, Deodato RM, Miguel JC, da Silva EF, Flores GL, Lewis-Ximenez LL. Usefulness of automated assays for detecting hepatitis B and C markers in dried blood spot samples. BMC Res Notes 2019; 12:523. [PMID: 31429797 PMCID: PMC6700985 DOI: 10.1186/s13104-019-4547-y] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/27/2019] [Accepted: 08/07/2019] [Indexed: 12/28/2022] Open
Abstract
Objective Dried blood spots (DBSs) can be used as an alternative to serum samples because they are easily collected and can be transported without refrigeration to reference laboratories for diagnosis. The present study was performed to evaluate the utility of electrochemiluminescence immunoassay “ECLIA” for anti-HCV, HBsAg and anti-HBc detection from DBS samples. Results Anti-HCV was detected in 103 DBS samples from 108 paired, positive serum and undetected in 364 DBS samples from 366 paired, negative specimens, giving a sensitivity of 95.4% and a specificity of 99.4%. HBsAg was detected in 67 DBS samples out of 71 positive, paired serum and was undetected among 295 DBS samples from 298 paired, negative specimens, giving a sensitivity and specificity of 94.4% and 99%, respectively. Anti-HBc was detected in 160 DBS samples from 185 paired, positive serum specimens and undetected in 349 DBS samples from 357 paired, negative serum specimens, giving a sensitivity of 86.5% and a specificity of 97.8%. Overall, the Kappa index indicated a high agreement between results obtained for the serum and DBS samples (k: 0.95, 0.93 and 0.86 for anti-HCV, HBsAg, anti-HBc, respectively). In conclusion, the ECLIA test could be used for detecting hepatitis B and C markers in DBS.
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Affiliation(s)
- Livia Melo Villar
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Hélio and Peggy Pereira Pavilion - Ground Floor - Office B09, FIOCRUZ Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil.
| | - Helena Medina Cruz
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Hélio and Peggy Pereira Pavilion - Ground Floor - Office B09, FIOCRUZ Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil
| | - Raissa Martins Deodato
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Hélio and Peggy Pereira Pavilion - Ground Floor - Office B09, FIOCRUZ Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil
| | - Juliana Custódio Miguel
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Hélio and Peggy Pereira Pavilion - Ground Floor - Office B09, FIOCRUZ Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil
| | - Elisangela Ferreira da Silva
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Hélio and Peggy Pereira Pavilion - Ground Floor - Office B09, FIOCRUZ Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil
| | - Geane Lopes Flores
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Hélio and Peggy Pereira Pavilion - Ground Floor - Office B09, FIOCRUZ Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil
| | - Lia Laura Lewis-Ximenez
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Hélio and Peggy Pereira Pavilion - Ground Floor - Office B09, FIOCRUZ Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, RJ, 210360-040, Brazil
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Coste M, De Sèze M, Diallo A, Carrieri MP, Marcellin F, Boyer S. Burden and impacts of chronic hepatitis B infection in rural Senegal: study protocol of a cross-sectional survey in the area of Niakhar (AmBASS ANRS 12356). BMJ Open 2019; 9:e030211. [PMID: 31320358 PMCID: PMC6661601 DOI: 10.1136/bmjopen-2019-030211] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
INTRODUCTION Though Senegal has one of the highest estimated prevalence rates of chronic hepatitis B virus (HBV) infection worldwide, epidemiological data in the general population are lacking and consequences of the infection remain undocumented. The ANRS-12356 AmBASS study aims at evaluating the health and socioeconomic burden of chronic HBV infection at the individual, household and population level. Its specific objectives are (1) to document the epidemiology of chronic HBV infection, including prevalence and risk factors; (2) to assess the acceptability of home-based testing and first clinic visit; (3) to investigate the repercussions of chronic HBV infection on living conditions; and (4) to estimate the public health impact of chronic HBV infection at the population level and the feasibility of a decentralised model of HBV test and treat. METHODS AND ANALYSIS This multidisciplinary cross-sectional survey includes a twofold data collection: (1) home-based screening using dried blood spot (DBS) sampling and collection of sociodemographic, economic and behavioural data, and (2) additional clinical and biological data collection in chronic HBV carriers at the first clinic visit. The prevalence of chronic HBV infection will be estimated in the general population and in key subgroups. Risk factors for HBV acquisition in children will be explored using case-control analysis. HBV burden will be assessed through comparisons of health and economic outcomes between households affected by the disease versus non-affected households. Last, an economic evaluation will assess costs and health benefits of scaling-up HBV care. ETHICS AND DISSEMINATION This study was approved by the Senegalese National Ethical Committee for Research in Health, and received authorisation from the Senegalese Ministry of Health and the French Commission on Information Technology and Liberties (Senegalese Protocol Number: SEN17/15). The study results will be presented in peer-review journals, international conferences and at a workshop with national stakeholders in order to contribute to the design of programmes to address the HBV pandemic. TRIAL REGISTRATION NUMBER NCT03215732; Pre-results.
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Affiliation(s)
- Marion Coste
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Economiques & Sociales de la Santé & Traitement de l'Information Médicale, Marseille, France
- ORS PACA, Observatoire Régional de la Santé Provence-Alpes-Côte d'Azur, Marseille, France
| | - Maëlle De Sèze
- Centre Européen de Sociologie et de Science Politique (CESSP-Paris, UMR 8209), Université Paris 1 Panthéon-Sorbonne, Paris, France
- School of Health and Related Research (ScHARR), University of Sheffield, Sheffield, UK
| | - Aldiouma Diallo
- Campus International IRD-UCAD de l'IRD, UMR VITROME, IRD-Université Aix Marseille, AP-HM, SSA, IHU-Méditerranée Infection, Dakar, Senegal
| | - Maria Patrizia Carrieri
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Economiques & Sociales de la Santé & Traitement de l'Information Médicale, Marseille, France
- ORS PACA, Observatoire Régional de la Santé Provence-Alpes-Côte d'Azur, Marseille, France
| | - Fabienne Marcellin
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Economiques & Sociales de la Santé & Traitement de l'Information Médicale, Marseille, France
- ORS PACA, Observatoire Régional de la Santé Provence-Alpes-Côte d'Azur, Marseille, France
| | - Sylvie Boyer
- Aix Marseille Univ, INSERM, IRD, SESSTIM, Sciences Economiques & Sociales de la Santé & Traitement de l'Information Médicale, Marseille, France
- ORS PACA, Observatoire Régional de la Santé Provence-Alpes-Côte d'Azur, Marseille, France
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Jackson K, Holgate T, Tekoaua R, Nicholson S, Littlejohn M, Locarnini S. Evaluation of dried blood spots for hepatitis B and D serology and nucleic acid testing. J Med Virol 2019; 94:642-648. [PMID: 30977903 DOI: 10.1002/jmv.25485] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/08/2019] [Revised: 03/21/2019] [Accepted: 04/07/2019] [Indexed: 12/11/2022]
Abstract
Areas with the highest burden of hepatitis B virus (HBV) infection are often low-middle-income countries with limited access to diagnosis due to isolation, affordability, and/or feasibility. Dried blood spots (DBSs) provide an alternative for remote areas where collection and transportation of serum is impractical. In this study, the application of DBS for serological and molecular detection of HBV and hepatitis D virus (HDV) was evaluated. Hepatitis B surface antigen was detected in 87 of 91 (95.6%) DBS. Seventeen of 21 (81%) had detectable HBeAg and 52 of 71 (73.2%) were anti-HBe positive. Anti-HD was detectable in 11 of 12 (91.6%) spiked control DBS after an initial failure to detect in patient DBS. HBV DNA was detected from 50 of 70 (71.4%) DBS with serum loads greater than 200 IU/mL in an in-house assay and 18 of 24 (75%) DBS with loads exceeding 389 IU/mL in a commercial assay. Using linear regression, HBV DNA loads from DBS were able to predict serum loads in 46 of 50 (92%) samples to within 1 log of actual serum load. HDV RNA was detected in 42 of 47 (89%) DBS with serum levels greater than 7200 IU/mL. DBSs are recommended for diagnosis of HBV, monitoring, and detection of high loads in pregnant women where peripheral blood testing remains unfeasible. Detection of HDV RNA from DBS may prove useful in endemic areas.
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Affiliation(s)
- Kathy Jackson
- Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital and WHO Regional Reference Laboratory for Hepatitis B/D, at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia
| | - Thomas Holgate
- Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital and WHO Regional Reference Laboratory for Hepatitis B/D, at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia
| | - Rosemary Tekoaua
- Tungaru Central Hospital, Ministry of Health and Medical Services, Tarawa, Republic of Kiribati
| | - Suellen Nicholson
- Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital and WHO Regional Reference Laboratory for Hepatitis B/D, at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia
| | - Margaret Littlejohn
- Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital and WHO Regional Reference Laboratory for Hepatitis B/D, at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia
| | - Stephen Locarnini
- Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital and WHO Regional Reference Laboratory for Hepatitis B/D, at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia
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Applicability of Oral Fluid and Dried Blood Spot for Hepatitis B Virus Diagnosis. Can J Gastroenterol Hepatol 2019; 2019:5672795. [PMID: 31058110 PMCID: PMC6463598 DOI: 10.1155/2019/5672795] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/13/2018] [Accepted: 02/03/2019] [Indexed: 12/12/2022] Open
Abstract
Hepatitis B virus (HBV) is one of the major causes of chronic liver disease worldwide; however most of individuals are not aware about the infection. Oral fluid and dried blood spot (DBS) samples may be an alternative to serum to HBV diagnosis to increase the access to diagnosis in remote areas or high-risk groups. The main objective of this review is to give an insight about the usefulness of oral fluid and DBS for detecting HBV markers. Several groups have evaluated the detection of HBsAg, anti-HBc, and anti-HBs markers in oral fluid and DBS samples demonstrating 13 to 100% of sensitivity and specificity according different groups, sample collectors, and diagnosis assays. In the same way, HBV DNA detection using oral fluid and DBS samples demonstrate different values of sensitivity according type of collection, studied group, extraction, and detection methods. Thus, serological and molecular diagnostic tests demonstrated good performance for detecting HBV using oral fluid and DBS according some characteristics and could be useful to increase the access to the diagnosis of HBV.
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Kenmoe S, Tagnouokam PAN, Nde CK, Mella-Tamko GF, Njouom R. Using dried blood spot for the detection of HBsAg and anti-HCV antibodies in Cameroon. BMC Res Notes 2018; 11:818. [PMID: 30446000 PMCID: PMC6240176 DOI: 10.1186/s13104-018-3931-3] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/19/2018] [Accepted: 11/14/2018] [Indexed: 12/24/2022] Open
Abstract
OBJECTIVE Dried blood spots (DBS) offer multiple benefits for collecting, storing and shipping whole blood samples. Our objective was to compare, for the first time in Africa, the performance of DBS with respect to plasma in the detection of Hepatitis B surface antigen (HBsAg) and antibodies to Hepatitis C Virus (anti-HCV) using Architect, Abbott Diagnostics. RESULTS DBS had a sensitivity of 99%, a specificity of 100%, a positive predictive value of 99%, a negative predictive value of 100% and a kappa index of 0.99 for the detection of HBsAg. For anti-HCV detection, the sensitivity, specificity, positive predictive value, negative predictive value and kappa index were 99%, 98%, 98%, 99%, and 0.97, respectively. This study confirms that DBS may be a reliable alternative specimen type for HBV and HCV diagnosis.
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Affiliation(s)
- Sebastien Kenmoe
- Virology Department, Centre Pasteur du Cameroun, 451 Rue 2005, Yaoundé 2, P.O.Box 1274, Yaounde, Cameroon
| | | | | | | | - Richard Njouom
- Virology Department, Centre Pasteur du Cameroun, 451 Rue 2005, Yaoundé 2, P.O.Box 1274, Yaounde, Cameroon
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Chevaliez S, Pawlotsky JM. New virological tools for screening, diagnosis and monitoring of hepatitis B and C in resource-limited settings. J Hepatol 2018; 69:916-926. [PMID: 29800630 DOI: 10.1016/j.jhep.2018.05.017] [Citation(s) in RCA: 69] [Impact Index Per Article: 9.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/28/2017] [Revised: 05/11/2018] [Accepted: 05/14/2018] [Indexed: 02/07/2023]
Abstract
Worldwide, the increasingly dominant model of laboratory testing is the centralised laboratory, in which automation of analytical processes increases, enabling the analysis of large numbers of samples at a relatively low cost. However, this trend does not fulfil the requirements for care of patients with chronic hepatitis B and C in resource-limited settings. Alternative models using point-of-care (POC) tests and dried blood spots (DBSs) are increasingly being considered for viral hepatitis screening, diagnosis and monitoring. POC tests are small devices providing qualitative and/or quantitative determination of viral antibodies and/or antigens. They can use original specimen matrices, such as oral fluid or blood collected from a fingerstick. POC tests are particularly useful for large-scale screening, and to improve access to care in regions where laboratory access is limited. New POC devices that detect and quantify viral nucleic acids are at the developmental stage. DBSs offer the main advantage of enabling storage of desiccated blood that can be easily transported to reference centres, where state-of-the-art molecular and serological diagnostic tests are available. However, standardisation and better automation of DBS handling are needed. Herein, we review alternatives to classical hepatitis B and C virological tests, examining POC tests and DBSs, as well as alternatives to nucleic acid testing. Innovations in testing approaches resulting from the availability of these new assays are also discussed.
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Affiliation(s)
- Stéphane Chevaliez
- National Reference Center for Viral Hepatitis B, C and D, Department of Virology, Hôpital Henri Mondor, Université Paris-Est, Créteil, France; INSERM U955, Créteil, France.
| | - Jean-Michel Pawlotsky
- National Reference Center for Viral Hepatitis B, C and D, Department of Virology, Hôpital Henri Mondor, Université Paris-Est, Créteil, France; INSERM U955, Créteil, France
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Flores GL, Cruz HM, Miguel JC, Potsch DV, Pilotto JH, Lewis-Ximenez LL, Lampe E, Villar LM. Assessing hepatitis B immunity using dried blood spot samples from HIV+ individuals. J Med Virol 2018; 90:1863-1867. [PMID: 30085359 DOI: 10.1002/jmv.25275] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/14/2017] [Accepted: 07/19/2018] [Indexed: 12/17/2022]
Abstract
This study aims to evaluate the utility of an optimized enzyme immunoassay (EIA) to detect and quantify antibodies against hepatitis B surface antibody (anti-HBs) in dried blood spots (DBSs) within the context of human immunodeficiency virus (HIV) status. Serum and DBS samples were obtained from 56 HIV+ and 99 HIV- patients and subjected to EIA for the detection of anti-HBs, where sample volume and cut off value were modified for DBS testing. Sensitivities of anti-HBs detection in DBS were 79.8% and 76.8% in HIV- and HIV+ subjects, respectively. Concordant results for anti-HBs in serum and DBS presented high mean CD8 cell counts, HIV viral load and optical density (OD) values of anti-HBs. Anti-HBs titers were significantly higher in serum, whether or not anti-HBs titers were detected in DBS. It was possible to detect anti-HBs in DBS as low as 17.4 and 27.3 IU/mL among HIV+ and HIV- subjects, respectively. In conclusion, DBS can be used to detect and quantify anti-HBs in HIV-infected individuals, which could increase access to diagnosis and vaccination.
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Affiliation(s)
- Geane Lopes Flores
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil
| | - Helena Medina Cruz
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil
| | | | - Denise Vigo Potsch
- Infectious Disease Ambulatory, Clementino Fraga Filho Hospital, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil
| | - José Henrique Pilotto
- Department of Sexual Transmitted Disease, Nova Iguaçu General Hospital, Nova Iguaçu, Rio de Janeiro, Brazil.,Molecular Immunology Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil
| | | | - Elisabeth Lampe
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil
| | - Livia Melo Villar
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil
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Scherbaum N, Timm J, Richter F, Bonnet U, Bombeck J, Lajos S, Specka M. Outcome of a hepatitis B vaccination program for clients of a drug consumption facility. J Clin Virol 2018; 106:28-32. [PMID: 30015286 DOI: 10.1016/j.jcv.2018.04.014] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/10/2018] [Revised: 04/16/2018] [Accepted: 04/21/2018] [Indexed: 12/21/2022]
Abstract
BACKGROUND Intravenous drug users (IDUs) are a risk group for hepatitis B. In Germany, the hepatitis B virus (HBV) vaccination rates in IDUs are low. OBJECTIVES In this study the implementation and success of HBV vaccination in a drug consumption facility (DCF) was evaluated. STUDY DESIGN Clients attending a DCF were asked regarding their HBV status. In case of no known HBV infection and no previous vaccination, clients interested in HBV vaccination were offered a HBV blood testing. HBV vaccination was administered to susceptible clients in months 0, 1, 6. Booster vaccinations were offered to clients without seroconversion (anti-HBs < 100 U/l). RESULTS 193 out of 364 clients reported on a known HBV infection or immunity after vaccination. 95 (55.6%) out of 171 eligible clients underwent a HBV serology. According to HBV serology 31 (32.6%) out of 95 clients were not susceptible for vaccination (mainly due to an unknown HBV infection). 47 (73.4%) out of 64 clients susceptible were administered 3 vaccinations. 10 clients received at least one further vaccination. For those showing up for testing (36 out of 47 clients) the seroconversion rate was 69.4% (> 100 IU/l) and 83.3% (> 10 IU/l), respectively. DISCUSSION Only a minority of clients of a DCF was susceptible for HBV vaccination. 47 out of 64 (73.4%) susceptible clients underwent at least three administrations of the vaccine, mostly resulting in seroconversion. Even in IDUs attending a DCF, a clientele with unstable social and health conditions, HBV vaccination can be carried out successfully.
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Affiliation(s)
- N Scherbaum
- LVR-Hospital Essen, Department of Psychiatry and Psychotherapy, Faculty of Medicine, University of Duisburg-Essen, Essen, Germany.
| | - J Timm
- Institute for Virology, University Hospital, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | | | - U Bonnet
- Department of Psychiatry, Psychotherapy, and Psychosomatic Medicine, Evangelisches Krankenhaus Castrop-Rauxel, Germany
| | | | - S Lajos
- LVR-Hospital Essen, Department of Psychiatry and Psychotherapy, Faculty of Medicine, University of Duisburg-Essen, Essen, Germany
| | - M Specka
- LVR-Hospital Essen, Department of Psychiatry and Psychotherapy, Faculty of Medicine, University of Duisburg-Essen, Essen, Germany
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Lange B, Cohn J, Roberts T, Camp J, Chauffour J, Gummadi N, Ishizaki A, Nagarathnam A, Tuaillon E, van de Perre P, Pichler C, Easterbrook P, Denkinger CM. Diagnostic accuracy of serological diagnosis of hepatitis C and B using dried blood spot samples (DBS): two systematic reviews and meta-analyses. BMC Infect Dis 2017; 17:700. [PMID: 29143672 PMCID: PMC5688450 DOI: 10.1186/s12879-017-2777-y] [Citation(s) in RCA: 67] [Impact Index Per Article: 8.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022] Open
Abstract
BACKGROUND Dried blood spots (DBS) are a convenient tool to enable diagnostic testing for viral diseases due to transport, handling and logistical advantages over conventional venous blood sampling. A better understanding of the performance of serological testing for hepatitis C (HCV) and hepatitis B virus (HBV) from DBS is important to enable more widespread use of this sampling approach in resource limited settings, and to inform the 2017 World Health Organization (WHO) guidance on testing for HBV/HCV. METHODS We conducted two systematic reviews and meta-analyses on the diagnostic accuracy of HCV antibody (HCV-Ab) and HBV surface antigen (HBsAg) from DBS samples compared to venous blood samples. MEDLINE, EMBASE, Global Health and Cochrane library were searched for studies that assessed diagnostic accuracy with DBS and agreement between DBS and venous sampling. Heterogeneity of results was assessed and where possible a pooled analysis of sensitivity and specificity was performed using a bivariate analysis with maximum likelihood estimate and 95% confidence intervals (95%CI). We conducted a narrative review on the impact of varying storage conditions or limits of detection in subsets of samples. The QUADAS-2 tool was used to assess risk of bias. RESULTS For the diagnostic accuracy of HBsAg from DBS compared to venous blood, 19 studies were included in a quantitative meta-analysis, and 23 in a narrative review. Pooled sensitivity and specificity were 98% (95%CI:95%-99%) and 100% (95%CI:99-100%), respectively. For the diagnostic accuracy of HCV-Ab from DBS, 19 studies were included in a pooled quantitative meta-analysis, and 23 studies were included in a narrative review. Pooled estimates of sensitivity and specificity were 98% (CI95%:95-99) and 99% (CI95%:98-100), respectively. Overall quality of studies and heterogeneity were rated as moderate in both systematic reviews. CONCLUSION HCV-Ab and HBsAg testing using DBS compared to venous blood sampling was associated with excellent diagnostic accuracy. However, generalizability is limited as no uniform protocol was applied and most studies did not use fresh samples. Future studies on diagnostic accuracy should include an assessment of impact of environmental conditions common in low resource field settings. Manufacturers also need to formally validate their assays for DBS for use with their commercial assays.
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Affiliation(s)
- Berit Lange
- Division of Infectious Diseases, Department of Medicine II, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Germany, Freiburg, Germany. .,Centre for Chronic Immunodeficiency, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Germany, Freiburg, Germany.
| | - Jennifer Cohn
- Department of Infectious Diseases, University of Pennsylvania, PA, Philadelphia, USA
| | | | - Johannes Camp
- Division of Infectious Diseases, Department of Medicine II, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Germany, Freiburg, Germany
| | | | - Nina Gummadi
- School of Medicine, Boston University, Boston, USA
| | - Azumi Ishizaki
- Global Hepatitis Programme, HIV Department, World Health Organization, Geneva, Switzerland
| | | | - Edouard Tuaillon
- Pathogenesis and Control of Chronic Infections UMR 1058 INSERM/Université Montpellier/Etablissement Français du Sang, INSERM, 34394, Montpellier Cedex 5, France.,Centre Hospitalier Universitaire (CHU) de Montpellier, Département de bactériologie-virologie, Montpellier, France
| | - Philippe van de Perre
- Pathogenesis and Control of Chronic Infections UMR 1058 INSERM/Université Montpellier/Etablissement Français du Sang, INSERM, 34394, Montpellier Cedex 5, France.,Centre Hospitalier Universitaire (CHU) de Montpellier, Département de bactériologie-virologie, Montpellier, France
| | - Christine Pichler
- Department of Medicine II, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany
| | - Philippa Easterbrook
- Global Hepatitis Programme, HIV Department, World Health Organization, Geneva, Switzerland
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Lange B, Roberts T, Cohn J, Greenman J, Camp J, Ishizaki A, Messac L, Tuaillon E, van de Perre P, Pichler C, Denkinger CM, Easterbrook P. Diagnostic accuracy of detection and quantification of HBV-DNA and HCV-RNA using dried blood spot (DBS) samples - a systematic review and meta-analysis. BMC Infect Dis 2017; 17:693. [PMID: 29143616 PMCID: PMC5688458 DOI: 10.1186/s12879-017-2776-z] [Citation(s) in RCA: 68] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
BACKGROUND The detection and quantification of hepatitis B (HBV) DNA and hepatitis C (HCV) RNA in whole blood collected on dried blood spots (DBS) may facilitate access to diagnosis and treatment of HBV and HCV infection in resource-poor settings. We evaluated the diagnostic performance of DBS compared to venous blood samples for detection and quantification of HBV-DNA and HCV-RNA in two systematic reviews and meta-analyses on the diagnostic accuracy of HBV DNA and HCV RNA from DBS compared to venous blood samples. METHODS We searched MEDLINE, Embase, Global Health, Web of Science, LILAC and Cochrane library for studies that assessed diagnostic accuracy with DBS. Heterogeneity was assessed and where appropriate pooled estimates of sensitivity and specificity were generated using bivariate analyses with maximum likelihood estimates and 95% confidence intervals. We also conducted a narrative review on the impact of varying storage conditions or different cut-offs for detection from studies that undertook this in a subset of samples. The QUADAS-2 tool was used to assess risk of bias. RESULTS In the quantitative synthesis for diagnostic accuracy of HBV-DNA using DBS, 521 citations were identified, and 12 studies met the inclusion criteria. Overall quality of studies was rated as low. The pooled estimate of sensitivity and specificity for HBV-DNA was 95% (95% CI: 83-99) and 99% (95% CI: 53-100), respectively. In the two studies that reported on cut-offs and limit of detection (LoD) - one reported a sensitivity of 98% for a cut-off of ≥2000 IU/ml and another reported a LoD of 914 IU/ml using a commercial assay. Varying storage conditions for individual samples did not result in a significant variation of results. In the synthesis for diagnostic accuracy of HCV-RNA using DBS, 15 studies met the inclusion criteria, and this included six additional studies to a previously published review. The pooled sensitivity and specificity was 98% (95% CI:95-99) and 98% (95% CI:95-99.0), respectively. Varying storage conditions resulted in a decrease in accuracy for quantification but not for reported positivity. CONCLUSIONS These findings show a high level of diagnostic performance for the use of DBS for HBV-DNA and HCV-RNA detection. However, this was based on a limited number and quality of studies. There is a need for development of standardized protocols by manufacturers on the use of DBS with their assays, as well as for larger studies on use of DBS conducted in different settings and with varying storage conditions.
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Affiliation(s)
- Berit Lange
- Division of Infectious Diseases, Department of Medicine II, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
- Centre for Chronic Immunodeficiency, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
| | | | - Jennifer Cohn
- Department of Infectious Diseases, University of Pennsylvania, Philadelphia, PA, USA
| | - Jamie Greenman
- Department of Infectious Diseases, University of Pennsylvania, Philadelphia, PA, USA
| | - Johannes Camp
- Division of Infectious Diseases, Department of Medicine II, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany
| | - Azumi Ishizaki
- Global Hepatitis Programme, HIV Department, World Health Organization, Geneva, Switzerland
| | - Luke Messac
- Department of Infectious Diseases, University of Pennsylvania, Philadelphia, PA, USA
| | - Edouard Tuaillon
- Pathogenesis and Control of Chronic Infections UMR 1058, INSERM/Université Montpellier/Etablissement Français du Sang, INSERM, 34394, Cedex 5, Montpellier, France
- Centre Hospitalier Universitaire (CHU) de Montpellier, département de bactériologie-virologie, Montpellier, France
| | - Philippe van de Perre
- Pathogenesis and Control of Chronic Infections UMR 1058, INSERM/Université Montpellier/Etablissement Français du Sang, INSERM, 34394, Cedex 5, Montpellier, France
- Centre Hospitalier Universitaire (CHU) de Montpellier, département de bactériologie-virologie, Montpellier, France
| | - Christine Pichler
- Division of Infectious Diseases, Department of Medicine II, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany
| | | | - Philippa Easterbrook
- Global Hepatitis Programme, HIV Department, World Health Organization, Geneva, Switzerland
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Gentile I, Zappulo E, Riccio MP, Binda S, Limauro R, Scuccimarra G, Borgia G, Bravaccio C. No evidence of congenital varicella zoster virus infection assessed through dried blood spot in children with autism spectrum disorders. Future Virol 2017. [DOI: 10.2217/fvl-2017-0039] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
Aim: Several authors have hypothesized an association between congenital viral infections and the onset of autism spectrum disorders (ASD). We aimed to assess the prevalence of congenital varicella zoster virus (VZV) infection in patients with ASD. Patients & methods: Congenital infection by VZV was evaluated in a cohort of 38 children with ASD and in 44 healthy controls. PCR for VZV-DNA performed on dried blood spots collected at birth. Results & conclusion: No VZV infection was detected in both groups. With the limitation of the small sample size of this study, the results are not in favor of a role of VZV in the etiology of ASD.
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Affiliation(s)
- Ivan Gentile
- Department of Clinical Medicine & Surgery, Section of Infectious Diseases, University of Naples “Federico II”, Naples, Italy
| | - Emanuela Zappulo
- Department of Clinical Medicine & Surgery, Section of Infectious Diseases, University of Naples “Federico II”, Naples, Italy
| | - Maria Pia Riccio
- Department of Medical Translational Science, University of Naples “Federico II”, Naples, Italy
| | - Sandro Binda
- Department of Biomedical Sciences for Health, University of Milan, Milan, Italy
| | | | | | - Guglielmo Borgia
- Department of Clinical Medicine & Surgery, Section of Infectious Diseases, University of Naples “Federico II”, Naples, Italy
| | - Carmela Bravaccio
- Department of Medical Translational Science, University of Naples “Federico II”, Naples, Italy
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Flores GL, Cruz HM, Potsch DV, May SB, Brandão-Mello CE, Pires MMA, Pilotto JH, Lewis-Ximenez LL, Lampe E, Villar LM. Evaluation of HBsAg and anti-HBc assays in saliva and dried blood spot samples according HIV status. J Virol Methods 2017; 247:32-37. [PMID: 28506632 DOI: 10.1016/j.jviromet.2017.05.004] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2016] [Revised: 04/22/2017] [Accepted: 05/09/2017] [Indexed: 02/07/2023]
Abstract
Influence of HIV status in HBV markers detection in saliva and dried blood spots (DBS) was not well established. This study aims to evaluate the performance of optimized commercial immunoassay for identifying HBsAg and anti-HBc in saliva and DBS according HIV status. A sum of 535 individuals grouped as HIV+, HBV+, HIV/HBV+ and HIV/HBV- were recruited where 347 and 188 were included for HBsAg and anti-HBc evaluation, respectively. Serum, DBS collected in Whatman 903 paper and saliva obtained using salivette device were analyzed using EIA. Increased sample volume and ROC curve analysis for cut off determination were used for DBS and saliva testing. HBsAg detection in saliva and DBS exhibited sensitivities of 80.9% and 85.6% and specificities of 86.8% and 96.3%. Sensitivity of anti-HBc in saliva and DBS were 82.4% and 76.9% and specificities in saliva and DBS were 96.9% and 91.7%. Low sensitivities were observed for HBsAg (62%) and anti-HBc (47%) detection in saliva of HIV/HBV+ individuals. OD values were also lower for HBsAg detection in DBS and saliva of HIV/HBV+ individuals compared to their serum samples. Statistical significance was found for sensitivities in HBsAg detection between saliva and DBS demonstrating high sensitivity for DBS specimens. In conclusion, HIV status or antiretroviral treatment appears to interfere in the performance of HBsAg and anti-HBc detection in DBS and saliva samples using the adapted commercial EIA.
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Affiliation(s)
- Geane Lopes Flores
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil
| | - Helena Medina Cruz
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil
| | - Denise Vigo Potsch
- Clementino Fraga Filho Hospital, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil
| | - Silvia Beatriz May
- Clementino Fraga Filho Hospital, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil
| | | | | | - Jose Henrique Pilotto
- Nova Iguaçu General Hospital & AIDS and Molecular Immunology Laboratory, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil
| | | | - Elisabeth Lampe
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil
| | - Livia Melo Villar
- Viral Hepatitis Laboratory, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil.
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BROUARD C, PILLONEL J, SOGNI P, CHOLLET A, LAZARUS JV, PASCAL X, BARIN F, JAUFFRET-ROUSTIDE M, the ANRS Coquelicot Survey Group. Hepatitis B virus in drug users in France: prevalence and vaccination history, ANRS-Coquelicot Survey 2011-2013. Epidemiol Infect 2017; 145:1-11. [PMID: 28100289 PMCID: PMC9507829 DOI: 10.1017/s0950268816003137] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2016] [Revised: 11/24/2016] [Accepted: 11/30/2016] [Indexed: 01/10/2023] Open
Abstract
People who use drugs (PWUD) are a key population for hepatitis B virus (HBV) vaccination and screening. We aimed to estimate the seroprevalence of HBs antigen (HBsAg) and self-reported HBV vaccination history in French PWUD attending harm reduction centres using data from the ANRS-Coquelicot multicentre survey conducted in 2011-2013 in 1718 PWUD. Self-fingerprick blood samples were collected on dried blood spots to detect the presence of HBsAg. HBsAg seroprevalence was estimated at 1·4% [95% confidence interval (CI) 0·8-2·5]. It varied between PWUD born in high (7·6%, 95% CI 2·7-19·1), moderate (2·2%, 95% CI 0·8-5·7) and low (0·7%, 95% CI 0·3-1·5) endemic zones. Factors independently associated with HBsAg carriage were being born in a moderate or high endemic zone or reporting precarious housing. Self-reported HBV vaccination history varied from 47·4% in high endemic zones, to 59·3% and 62·6% for moderate and low endemic zones, respectively. Our results suggest that drug use plays a small and substantial role, respectively, in HBsAg carriage in PWUD born in high/moderate and low endemic zones.
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Affiliation(s)
- C. BROUARD
- Santé publique France, Direction des Maladies Infectieuses, Saint-Maurice, France
| | - J. PILLONEL
- Santé publique France, Direction des Maladies Infectieuses, Saint-Maurice, France
| | - P. SOGNI
- Université Paris Descartes, Paris, France
- Inserm U1223, Institut Pasteur, France
- Assistance Publique des Hôpitaux de Paris, Hôpital Cochin, Service d'Hépatologie, Paris, France
| | - A. CHOLLET
- Cermes3, Inserm U988, UMR CNRS 8211, Université Paris Descartes, EHESS, Paris, France
| | - J. V. LAZARUS
- Centre for Health and Infectious Disease Research and WHO Collaborating Centre on HIV and Viral Hepatitis, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark
- ISGlobal, Hospital Clínic, University of Barcelona, Barcelona, Spain
| | - X. PASCAL
- Cermes3, Inserm U988, UMR CNRS 8211, Université Paris Descartes, EHESS, Paris, France
| | - F. BARIN
- Inserm U966, Centre National de Référence du VIH, CHU Bretonneau, Université François-Rabelais, Tours, France
| | - M. JAUFFRET-ROUSTIDE
- Santé publique France, Direction des Maladies Infectieuses, Saint-Maurice, France
- Cermes3, Inserm U988, UMR CNRS 8211, Université Paris Descartes, EHESS, Paris, France
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Dry Blood Spots a Reliable Method for Measurement of Hepatitis B Viral Load in Resource-Limited Settings. PLoS One 2016; 11:e0166201. [PMID: 27820845 PMCID: PMC5098817 DOI: 10.1371/journal.pone.0166201] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/19/2016] [Accepted: 10/24/2016] [Indexed: 12/22/2022] Open
Abstract
Background & Aims Hepatitis B virus (HBV) quantification is essential in the management of chronic hepatitis B, both to determine treatment eligibility and in the monitoring of treatment effect. This test, however, is rarely available in resource-limited settings due to high costs and stringent requirements for shipment and storage of plasma. Dried Blood Spots (DBS) can be a convenient alternative to plasma, but its use for HBV monitoring has not been investigated under real-life conditions in Africa. Methods The performance of DBS in HBV quantification was investigated using a modified commercial test (Abbott RealTime HBV assay). Paired DBS and plasma samples were collected from an HBV positive cohort in Addis Ababa, Ethiopia. DBS were stored at ambient temperature for 4–39 days before shipment to the laboratory. Results Twenty-six paired samples were selected covering the total range of quantification, from 2.14 log IU/ml to >7 log IU/ml. HBV was detected in 21 of 21 (100%) DBS from patients with a corresponding plasma viral load above 2.70 log IU/ml. The mean difference between plasma and DBS was 0.59 log IU/ml, and the correlation was strong (R2 = 0.92). In stability studies there was no significant change in DBS viral load after storage at room temperature for up to 12 weeks. Conclusions This study suggests that DBS can be a feasible and reliable alternative to plasma for quantification of HBV in resource-limited settings. DBS can expand access to antiviral treatment for patients in low- and middle-income countries.
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Allain JP, Opare-Sem O. Screening and diagnosis of HBV in low-income and middle-income countries. Nat Rev Gastroenterol Hepatol 2016; 13:643-653. [PMID: 27625189 DOI: 10.1038/nrgastro.2016.138] [Citation(s) in RCA: 49] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Abstract
HBV testing and diagnosis of HBV-related liver disease in low-income and middle-income countries differs substantially from that in developed countries in terms of access to resources and expensive technologies requiring highly specialized staff. For identification and classification of HBV infection, genomic amplification methods to detect and quantify HBV DNA are often nonexistent or available only in central laboratories of major cities. When samples from peripheral locations do arrive, delays in receiving results generate loss to follow-up. Testing is often limited to measurement of hepatitis B surface antigen (HBsAg), alanine aminotransferase levels, aspartate aminotransferase to platelet ratio index and hepatitis B e antigen (HBeAg) to determine indications for antiviral therapy (AVT). Utilization of AVT is limited by cost and availability, particularly when patients are not covered by health insurance. The natural history of HBV infection is influenced by genotypes B and C in East Asia, where decades of immune tolerance have led to mostly vertical transmission; in sub-Saharan Africa, where genotypes A1 and E predominate, infection is transmitted horizontally between young children, followed by a nonreplicative phase. In both regions, cirrhosis and hepatocellular carcinoma are common and would be considerably ameliorated by AVT. Implementation of the HBV vaccine since the 1990s in Asia and 2000s in Africa has decreased the incidence of HBV, but vaccine failure and insufficiently effective prevention remain concerning issues.
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Affiliation(s)
- Jean-Pierre Allain
- Department of Haematology, University of Cambridge, Science Village, Chesterford Research Park, Little Chesterford CB10 1XL, UK
| | - Ohene Opare-Sem
- Department of Medicine, Kwame Nkrumah University of Science and Technology, University Post Office, Kumasi, Ghana
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Mössner BK, Staugaard B, Jensen J, Lillevang ST, Christensen PB, Holm DK. Dried blood spots, valid screening for viral hepatitis and human immunodeficiency virus in real-life. World J Gastroenterol 2016; 22:7604-7612. [PMID: 27672281 PMCID: PMC5011674 DOI: 10.3748/wjg.v22.i33.7604] [Citation(s) in RCA: 58] [Impact Index Per Article: 6.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/29/2016] [Revised: 06/27/2016] [Accepted: 08/01/2016] [Indexed: 02/06/2023] Open
Abstract
AIM To detect chronic hepatitis B (CHB), chronic hepatitis C (CHC) and human immunodeficiency virus (HIV) infections in dried blood spot (DBS) and compare these samples to venous blood sampling in real-life.
METHODS We included prospective patients with known viral infections from drug treatment centers, a prison and outpatient clinics and included blood donors as negative controls. Five drops of finger capillary blood were spotted on filter paper, and a venous blood sample was obtained. The samples were analyzed for HBsAg, anti-HBc, anti-HBs, anti-HCV, and anti-HIV levels as well as subjected to a combined nucleic acid test (NAT) for HBV DNA, HCV RNA and HIV RNA.
RESULTS Samples from 404 subjects were screened (85 CHB, 116 CHC, 114 HIV and 99 blood donors). DBS had a sensitivity of > 96% and a specificity of > 98% for the detection of all three infections. NAT testing did not improve sensitivity, but correctly classified 95% of the anti-HCV-positive patients with chronic and past infections. Anti-HBc and anti-HBS showed low sensitivity in DBS (68% and 42%).
CONCLUSION DBS sampling, combined with an automated analysis system, is a feasible screening method to diagnose chronic viral hepatitis and HIV infections outside of the health care system.
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Barnhart M. A Convenient Truth: Cost of Medications Need Not Be a Barrier to Hepatitis B Treatment. GLOBAL HEALTH: SCIENCE AND PRACTICE 2016; 4:186-90. [PMID: 27353613 PMCID: PMC4982244 DOI: 10.9745/ghsp-d-16-00128] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Download PDF] [Subscribe] [Scholar Register] [Indexed: 12/30/2022]
Affiliation(s)
- Matthew Barnhart
- Global Health, Science and Practice, Associate Editor,
Washington, DC, USA
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45
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Villar LM, Cruz HM, Barbosa JR, Bezerra CS, Portilho MM, Scalioni LDP. Update on hepatitis B and C virus diagnosis. World J Virol 2015; 4:323-42. [PMID: 26568915 PMCID: PMC4641225 DOI: 10.5501/wjv.v4.i4.323] [Citation(s) in RCA: 53] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/03/2015] [Revised: 09/25/2015] [Accepted: 10/23/2015] [Indexed: 02/05/2023] Open
Abstract
Viral hepatitis B and C virus (HBV and HCV) are responsible for the most of chronic liver disease worldwide and are transmitted by parenteral route, sexual and vertical transmission. One important measure to reduce the burden of these infections is the diagnosis of acute and chronic cases of HBV and HCV. In order to provide an effective diagnosis and monitoring of antiviral treatment, it is important to choose sensitive, rapid, inexpensive, and robust analytical methods. Primary diagnosis of HBV and HCV infection is made by using serological tests for detecting antigens and antibodies against these viruses. In order to confirm primary diagnosis, to quantify viral load, to determine genotypes and resistance mutants for antiviral treatment, qualitative and quantitative molecular tests are used. In this manuscript, we review the current serological and molecular methods for the diagnosis of hepatitis B and C.
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Gueye SB, Diop-Ndiaye H, Lo G, Mintsa S, Guindo I, Dia A, Sow-Sall A, Gaye-Diallo A, Mboup S, Touré-Kane C. HBV carriage in children born from HIV-seropositive mothers in Senegal: The need of birth-dose HBV vaccination. J Med Virol 2015; 88:815-9. [PMID: 26488892 DOI: 10.1002/jmv.24409] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 10/18/2015] [Indexed: 02/06/2023]
Abstract
Hepatitis B is a major public health problem in Senegal, a country with high prevalence and a transmission occurring mainly during infancy. Only, one 6-8 weeks vaccination campaign was initiated in 2005 and it was part of the expanded program of immunization. The aim of this study was to determine the prevalence of HBsAg in children born from HIV-seropositive mothers by using dried blood specimens. Specimens were collected between July 2007 and November 2012 from children aged 2-48 weeks in Dakar and decentralized sites working on HIV mother-to-child transmission prevention. HBsAg detection was performed using Architect HBsAg Qualitative II kit (Abbott Diagnostics, Ireland) and for all reactive samples confirmation was done using Architect HBsAg Qualitative II Confirmatory kit (Abbott Diagnostics, Ireland). Nine hundred thirty samples were collected throughout the country with 66% out of Dakar, the capital city. The median age was 20 weeks and 88% of children were less than 1 year of age with a sex ratio of 1.27 in favor of boys. HBsAg was detected in 28 cases giving a global prevalence of 3%. According to age, HBsAg prevalences were 5.1% for children less than 6 weeks, 4.1% and 4.6%, respectively, for those aged 12-18 weeks and 18-24 weeks of age. The HIV prevalence was 2.6% with no HIV/HBV co-infection. This study showed a high rate of HBV infection in children under 24 months, highlighting the need to promote birth-dose HBV vaccination as recommended by WHO.
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Affiliation(s)
- Sokhna Bousso Gueye
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Halimatou Diop-Ndiaye
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Gora Lo
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Sandrine Mintsa
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Ibrahima Guindo
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Aminata Dia
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Amina Sow-Sall
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Aissatou Gaye-Diallo
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Souleymane Mboup
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
| | - Coumba Touré-Kane
- Laboratoire Bactériologie-Virologie, CHU Le Dantec, Université Cheikh Anta Diop, Dakar-Sénégal, Dakar, Senegal
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Calisti G, Muhindo R, Boum Y, Wilson LA, Foster GM, Geretti AM, Bhagani S. Epidemiology of HBV infection in a cohort of Ugandan HIV-infected patients and rate and pattern of lamivudine-resistant HBV infection in patients receiving antiretroviral therapy. Trans R Soc Trop Med Hyg 2015; 109:723-9. [PMID: 26386408 DOI: 10.1093/trstmh/trv077] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/11/2015] [Accepted: 08/21/2015] [Indexed: 12/15/2022] Open
Abstract
BACKGROUND Many HIV-infected patients in sub-Saharan Africa are not routinely screened for hepatitis B virus (HBV) infection and are on antiretroviral therapy (ART) regimens containing only lamivudine as anti-HBV active drug. METHODS In 2009-2011, we screened for hepatitis B surface antigen (HBsAg) in 2820 HIV-infected adults patients at the Mbarara Hospital Uganda and investigated risk factors for HBV infection. Using samples of dried plasma or blood spots, we tested for HBV viral load and HBV drug resistance mutations in all HBsAg-positive patients on ART for ≥ 12 months. RESULTS In this study, 109 patients tested HBsAg positive (3.9%; 109/2820). HBsAg-positive patients were more likely to have had >4 lifetime sexual partners (p<0.01). Of the 55 HBsAg-positive patients on ART for ≥ 12 months, 53 were only on lamivudine as anti-HBV active drug and two were on tenofovir and lamivudine. HBV-DNA was detected in 30 patients (54.5%; 30/55), all on lamivudine-monotherapy. Of the 23 patients in whom HBV-DNA sequencing was successful, 17 had lamivudine-resistant HBV strains harbouring rtM204V/I mutations accompanied by secondary/compensatory mutations. CONCLUSIONS Our study suggests that sexual transmission may represent a major mode of spread of HBV in southwest Uganda and confirms the importance of screening for HBV and of using ART regimens containing tenofovir in HIV/HBV co-infected patients.
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Affiliation(s)
- Giorgio Calisti
- Department of Virology, Royal Free London NHS Foundation Trust, London, UK
| | - Rose Muhindo
- Faculty of Medicine, Mbarara University of Science and Technology, Mbarara, Uganda
| | - Yap Boum
- Epicentre Mbarara Research Base, Mbarara, Uganda
| | - Laurence A Wilson
- Faculty of Medicine, Mbarara University of Science and Technology, Mbarara, Uganda
| | - Geraldine M Foster
- Institute of Infection and Global Health, University of Liverpool, Liverpool, UK
| | - Anna Maria Geretti
- Institute of Infection and Global Health, University of Liverpool, Liverpool, UK
| | - Sanjay Bhagani
- Department of HIV Medicine, Royal Free London NHS Foundation Trust, London, UK
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48
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Vinikoor MJ, Zürcher S, Musukuma K, Kachuwaire O, Rauch A, Chi BH, Gorgievski M, Zwahlen M, Wandeler G. Hepatitis B viral load in dried blood spots: A validation study in Zambia. J Clin Virol 2015; 72:20-4. [PMID: 26356987 DOI: 10.1016/j.jcv.2015.08.019] [Citation(s) in RCA: 22] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2015] [Revised: 08/29/2015] [Accepted: 08/31/2015] [Indexed: 11/17/2022]
Abstract
BACKGROUND Access to hepatitis B viral load (VL) testing is poor in sub-Saharan Africa (SSA) due to economic and logistical reasons. OBJECTIVES To demonstrate the feasibility of testing dried blood spots (DBS) for hepatitis B virus (HBV) VL in a laboratory in Lusaka, Zambia, and to compare HBV VLs between DBS and plasma samples. STUDY DESIGN Paired plasma and DBS samples from HIV-HBV co-infected Zambian adults were analyzed for HBV VL using the COBAS AmpliPrep/COBAS TaqMan HBV test (Version 2.0) and for HBV genotype by direct sequencing. We used Bland-Altman analysis to compare VLs between sample types and by genotype. Logistic regression analysis was conducted to assess the probability of an undetectable DBS result by plasma VL. RESULTS Among 68 participants, median age was 34 years, 61.8% were men, and median plasma HBV VL was 3.98logIU/ml (interquartile range, 2.04-5.95). Among sequenced viruses, 28 were genotype A1 and 27 were genotype E. Bland-Altman plots suggested strong agreement between DBS and plasma VLs. DBS VLs were on average 1.59logIU/ml lower than plasma with 95% limits of agreement of -2.40 to -0.83log IU/ml. At a plasma VL ≥2,000IU/ml, the probability of an undetectable DBS result was 1.8% (95% CI: 0.5-6.6). At plasma VL ≥20,000IU/ml this probability reduced to 0.2% (95% CI: 0.03-1.7). CONCLUSIONS In a Zambian laboratory, we observed strong agreement between DBS and plasma VLs and high sensitivity in DBS at plasma VL ≥2,000IU/ml. As HBV treatment expands, DBS could increase access to HBV VL testing and care in SSA settings.
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Affiliation(s)
- Michael J Vinikoor
- Department of Medicine, University of Alabama at Birmingham, USA; Centre for Infectious Disease Research in Zambia, Lusaka, Zambia; School of Medicine, University of Zambia, Lusaka, Zambia.
| | - Samuel Zürcher
- Institute of Infectious Diseases, University of Bern, Switzerland
| | - Kalo Musukuma
- Centre for Infectious Disease Research in Zambia, Lusaka, Zambia; School of Medicine, University of Zambia, Lusaka, Zambia
| | - Obert Kachuwaire
- Centre for Infectious Disease Research in Zambia, Lusaka, Zambia
| | - Andri Rauch
- Department of Infectious Diseases, University Hospital Bern, Switzerland
| | - Benjamin H Chi
- Department of Obstetrics and Gynecology, University of North Carolina at Chapel Hill, USA
| | - Meri Gorgievski
- Institute of Infectious Diseases, University of Bern, Switzerland
| | - Marcel Zwahlen
- Institute of Social and Preventive Medicine, University of Bern, Switzerland
| | - Gilles Wandeler
- Department of Infectious Diseases, University Hospital Bern, Switzerland; Institute of Social and Preventive Medicine, University of Bern, Switzerland; Department of Infectious Diseases, University of Dakar, Senegal
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49
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Abstract
Viral hepatitis claims one million lives each year. Scaling up treatment for hepatitis B and C in resource-limited settings is not possible without access to reliable diagnostic tools. This article gives an overview of current technologies and the pipeline for easy-to-use assays for serological and virological analyses, which can be performed at the site of patient care ('point-of-care assays'). Furthermore, the utility of dried blood spots for hepatitis B and C viral load testing is discussed. In addition to simple and reliable diagnostics, there is a need for a sustainable funding scheme and generic production of antiviral drugs to reduce the burden of viral hepatitis worldwide.
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Affiliation(s)
- A Johannessen
- Centre for Imported and Tropical Diseases, Oslo University HospitalOslo, Norway,
Correspondence: Dr A. Johannessen, Centre for Imported and Tropical Diseases, Oslo University Hospital, PO Box 4956 Nydalen, 0424 Oslo, Norway. E-mail:
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50
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Almeida RWD, Espírito-Santo MP, Sousa PSF, Almeida AJD, Lampe E, Lewis-Ximenez LL. Hepatitis B virus DNA stability in plasma samples under short-term storage at 42°C. ACTA ACUST UNITED AC 2015; 48:553-6. [PMID: 25790101 PMCID: PMC4470315 DOI: 10.1590/1414-431x20144040] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2014] [Accepted: 11/11/2014] [Indexed: 11/22/2022]
Abstract
We evaluated the stability of hepatitis B virus (HBV) DNA in plasma samples stored at
42°C for external quality assessment (EQA) panels of viral load. To assess the
stability of plasma samples containing different concentrations of HBV DNA, serial
dilutions of HBV-infected samples with a viral load of 6.40 log(10) IU/mL
were made to yield viral loads of 5, 4, and 3 log(10) IU/mL. These were
incubated at 42°C for up to 7 days and then frozen at -70°C. Viral load testing for
HBV DNA was performed for all samples using COBAS¯
AmpliPrep/COBAS¯ TaqMan¯ HBV Test (v.2.0, Roche,
Switzerland). Results were compared with fresh frozen plasma samples as a benchmark
to establish acceptable measurements on the days following sample collection.
Although the results of this study demonstrated a decrease in HBV DNA viral load
ranging from 0.005 to 0.30 log(10) IU/mL after storage at 42°C for up to 7
days, these values did not exceed 0.5 log(10), which is the estimated
intra-assay variation for molecular tests. Thus, the insignificant decrease in viral
load suggests that shipment of HBV in plasma samples at temperatures of up to 42°C is
permissible if they are frozen within 7 days.
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Affiliation(s)
- R W de Almeida
- Laboratório de Hepatites Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
| | - M P Espírito-Santo
- Laboratório de Hepatites Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
| | - P S F Sousa
- Laboratório de Hepatites Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
| | - A J de Almeida
- Laboratório de Hepatites Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
| | - E Lampe
- Laboratório de Hepatites Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
| | - L L Lewis-Ximenez
- Laboratório de Hepatites Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
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