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Lok J, Harris JM, Carey I, Agarwal K, McKeating JA. Assessing the virological response to direct-acting antiviral therapies in the HBV cure programme. Virology 2025; 605:110458. [PMID: 40022943 DOI: 10.1016/j.virol.2025.110458] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2024] [Revised: 01/16/2025] [Accepted: 02/20/2025] [Indexed: 03/04/2025]
Abstract
Hepatitis B virus (HBV) is a global health problem with over 250 million people affected worldwide. Nucleos(t)ide analogues remain the standard of care and suppress production of progeny virions; however, they have limited effect on the viral transcriptome and long-term treatment is associated with off-target toxicities. Promising results are emerging from clinical trials and several drug classes have been evaluated, including capsid assembly modulators and RNA interfering agents. Whilst peripheral biomarkers are used to monitor responses and define treatment endpoints, they fail to reflect the full reservoir of infected hepatocytes. Given these limitations, consideration should be given to the merits of sampling liver tissue, especially in the context of clinical trials. In this review article, we will discuss methods for profiling HBV in liver tissue and their value to the HBV cure programme.
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Affiliation(s)
- James Lok
- Institute of Liver Studies, King's College Hospital, London, SE5 9RS, United Kingdom.
| | - James M Harris
- Nuffield Department of Medicine, University of Oxford, OX3 7FZ, United Kingdom
| | - Ivana Carey
- Institute of Liver Studies, King's College Hospital, London, SE5 9RS, United Kingdom
| | - Kosh Agarwal
- Institute of Liver Studies, King's College Hospital, London, SE5 9RS, United Kingdom
| | - Jane A McKeating
- Nuffield Department of Medicine, University of Oxford, OX3 7FZ, United Kingdom; Chinese Academy of Medical Sciences Oxford Institute, University of Oxford, Oxford, United Kingdom
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2
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Lok J, Dusheiko G, Carey I, Agarwal K. Review article: novel biomarkers in hepatitis B infection. Aliment Pharmacol Ther 2022; 56:760-776. [PMID: 35770458 DOI: 10.1111/apt.17105] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/02/2022] [Revised: 06/08/2022] [Accepted: 06/12/2022] [Indexed: 12/09/2022]
Abstract
BACKGROUND Chronic hepatitis B remains a global health problem with an estimated 296 million people affected worldwide. Individuals are at risk of serious complications such as cirrhosis and hepatocellular carcinoma and accurately predicting these clinical endpoints has proven difficult. However, several viral biomarkers have recently been developed, including quantitative HBV surface antigen (qHBsAg), hepatitis B RNA (HBV RNA) and core-related antigen (HBcrAg), and shown promise in a range of clinical settings. AIMS To critically appraise these novel biomarkers, exploring their potential uses, availability of assays and areas for future development. METHODS We performed a literature search of PubMed, identifying articles published in the field of hepatitis B biomarkers between 2010 and 2022. RESULTS Novel biomarkers such as HBcrAg, HBV RNA and qHBsAg may be useful in predicting treatment outcomes, stratifying the risk of future complications and estimating off-treatment viral reactivation. Furthermore, HBV RNA and HBcrAg titres may accurately reflect cccDNA transcriptional activity, and this is particularly informative in the context of nucleoside analogue therapy. On a cautionary note, most studies have been performed in Caucasian or Asian populations, and methods for detecting HBV RNA lack standardisation. CONCLUSION Novel viral biomarkers have the potential to provide additional insights into the natural history of infection and allow a more bespoke, cost-effective framework of care. However, access remains limited, and further efforts are needed to validate their use in ethnically diverse populations, confirm predictive cut-off values, and establish their role in the era of novel antiviral therapies.
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Affiliation(s)
- James Lok
- Institute of Liver Studies, King's College Hospital, London, UK
| | | | - Ivana Carey
- Institute of Liver Studies, King's College Hospital, London, UK
| | - Kosh Agarwal
- Institute of Liver Studies, King's College Hospital, London, UK
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3
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Chung GE, Kim JY, Shin H, Hong JH, Hur MH, Cho H, Park MK, Choi NR, Kim J, Lee YB, Cho EJ, Yu SJ, Kim YJ, Yoon JH, Lee JH. Correlation between Results of Semi-Quantitative and Quantitative Tests for Hepatitis B Virus Surface Antigen among Patients Achieving Viral Suppression with Antiviral Treatment. Diagnostics (Basel) 2022; 12:diagnostics12071757. [PMID: 35885659 PMCID: PMC9317496 DOI: 10.3390/diagnostics12071757] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2022] [Revised: 07/13/2022] [Accepted: 07/19/2022] [Indexed: 11/16/2022] Open
Abstract
Background: Hepatitis B virus (HBV) infection remains a threat to global public health. Serum hepatitis B surface antigen (HBsAg) has been used in screening for HBV infection. Quantitative HBsAg assays are useful for monitoring the natural history of HBV infection and its response to therapy. The aim of this study was to determine the relationship between quantitative (qHBsAg; IU/mL) and semi-quantitative (sqHBsAg; signal-to-cutoff ratio [S/Co]) HBsAg titers in patients with chronic hepatitis B (CHB). Methods: We retrospectively included 284 samples with HBV DNA < 20 IU/mL from patients who had simultaneous qHBsAg (using electrochemiluminescence assay) and sqHBsAg tests. Patients were grouped according to their serum HBV-envelope antigen (HBeAg) status (HBeAg-negative, n = 239 and HBeAg-positive, n = 45). The Spearman test was used to analyze the correlation between the quantitative and semi-quantitative assays. Results: There was a significant linear correlation between sqHBsAg and qHBsAg in the HBeAg-negative patients (qHBsAg [IU/mL] = 0.0094 × sqHBsAg [S/Co]1.323; adjusted R2 = 0.8445; p < 0.001). There was a substantial hook effect in the assays from the HBeAg-positive patients, so we performed a stratified analysis according to qHBsAg <1000 IU/mL or ≥1000 IU/mL and found a significant positive linear correlation between sqHBsAg S/Co and qHBsAg (qHBsAg [IU/mL] = 0.072 × sqHBsAg [S/Co]1.331; adjusted R2 = 0.7878; p < 0.001) in HBeAg-positive patients with qHBsAg titers of <1000 IU/mL and a significant negative correlation in HBeAg-positive patients with qHBsAg titers of ≥1000 IU/mL (qHBsAg [IU/mL] = 8.987 × 1014 × sqHBsAg [S/Co]−3.175; adjusted R2 = 0.6350; p < 0.001). Conclusions: There was a highly linear, positive correlation between qHBsAg and sqHBsAg in HBeAg-negative CHB patients. The hook effect led to a negative correlation in HBeAg-positive CHB patients with qHBsAg titers ≥1000 IU/mL.
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Affiliation(s)
- Goh Eun Chung
- Department of Internal Medicine, Healthcare Research Institute, Gangnam Healthcare Center, Seoul National University Hospital, Seoul 03080, Korea;
| | - Ju Yeon Kim
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Hyunjae Shin
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Ji Hoon Hong
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Moon Haeng Hur
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Heejin Cho
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Min Kyung Park
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Na Ryung Choi
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Jihye Kim
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Yun Bin Lee
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Eun Ju Cho
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Su Jong Yu
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Yoon Jun Kim
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Jung-Hwan Yoon
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
| | - Jeong-Hoon Lee
- Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 03080, Korea; (J.Y.K.); (H.S.); (J.H.H.); (M.H.H.); (H.C.); (M.K.P.); (N.R.C.); (J.K.); (Y.B.L.); (E.J.C.); (S.J.Y.); (Y.J.K.); (J.-H.Y.)
- Correspondence: ; Tel.: +82-2-2072-2228
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Hepatitis B surface antigen seroclearance: Immune mechanisms, clinical impact, importance for drug development. J Hepatol 2020; 73:409-422. [PMID: 32333923 DOI: 10.1016/j.jhep.2020.04.013] [Citation(s) in RCA: 102] [Impact Index Per Article: 20.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/04/2019] [Revised: 04/03/2020] [Accepted: 04/07/2020] [Indexed: 12/16/2022]
Abstract
HBsAg seroclearance occurs rarely in the natural history of chronic hepatitis B (CHB) infection and is associated with improved clinical outcomes. Many factors are associated with HBsAg seroconversion, including immune and viral factors. However, the immune mechanisms associated with HBsAg seroclearance are still difficult to elucidate. HBsAg seroclearance is the ideal aim of HBV treatment. Unfortunately, this goal is rarely achieved with current treatments. Understanding the mechanisms of HBsAg loss appears to be important for the development of curative HBV treatments. While studies from animal models give insights into the potential immune mechanisms and interactions occurring between the immune system and HBsAg, they do not recapitulate all features of CHB in humans and are subject to variability due to their complexity. In this article, we review recent studies on these immune factors, focusing on their influence on CHB progression and HBsAg seroconversion. These data provide new insights for the development of therapeutic approaches to partially restore the anti-HBV immune response. Targeting HBsAg will ideally relieve the immunosuppressive effects on the immune system and help to restore anti-HBV immune responses.
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5
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Al-Sadeq DW, Taleb SA, Zaied RE, Fahad SM, Smatti MK, Rizeq BR, Al Thani AA, Yassine HM, Nasrallah GK. Hepatitis B Virus Molecular Epidemiology, Host-Virus Interaction, Coinfection, and Laboratory Diagnosis in the MENA Region: An Update. Pathogens 2019; 8:63. [PMID: 31083509 PMCID: PMC6630671 DOI: 10.3390/pathogens8020063] [Citation(s) in RCA: 25] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/30/2019] [Revised: 04/18/2019] [Accepted: 05/08/2019] [Indexed: 02/07/2023] Open
Abstract
Hepatitis B virus (HBV) is an enveloped partial double-stranded DNA virus that can cause acute and chronic hepatitis. According to the World Health Organization (WHO) and the Centers for Disease Control and Prevention (CDC), 257 million people are living with HBV. Moreover, 20,900 acute hepatitis B cases were reported in 2016. Hepatitis B is highly prevalent in the African, Western Pacific, Eastern Mediterranean, South-East Asia, and European regions, respectively. Due to the high mutational rate of HBV and lack of reverse transcriptase proofreading activity, ten different genotypes with different geographical distributions have been identified. HBV pathogenesis and severity of infection depend on several host and viral factors, particularly, the genetic variability of both the host and virus. Although HBV infection is a global health concern, there is a lack of adequate studies and reports in the Middle East and North Africa (MENA) region. Here, we provide a review on HBV epidemiology, pathogenesis, host-pathogen interactions, coinfection with selected viruses, and laboratory diagnosis, focusing on studies conducted in the MENA region to determine the current situation of the HBV infection and outline the future study areas.
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Affiliation(s)
- Duaa W Al-Sadeq
- Biomedical Research Center, Qatar University, Doha 2713, Qatar.
- Biomedical Science Department, College of Health Sciences, Qatar University, Doha 2713, Qatar.
| | - Sara A Taleb
- Biomedical Science Department, College of Health Sciences, Qatar University, Doha 2713, Qatar.
| | - Roan E Zaied
- Biomedical Science Department, College of Health Sciences, Qatar University, Doha 2713, Qatar.
| | - Sara M Fahad
- Biomedical Research Center, Qatar University, Doha 2713, Qatar.
| | - Maria K Smatti
- Biomedical Research Center, Qatar University, Doha 2713, Qatar.
| | - Balsam R Rizeq
- Biomedical Research Center, Qatar University, Doha 2713, Qatar.
- Department of Biological and Environmental Sciences, College of Arts & Sciences, Qatar University, Doha 2713, Qatar.
| | - Asmaa A Al Thani
- Biomedical Research Center, Qatar University, Doha 2713, Qatar.
- Biomedical Science Department, College of Health Sciences, Qatar University, Doha 2713, Qatar.
| | - Hadi M Yassine
- Biomedical Research Center, Qatar University, Doha 2713, Qatar.
| | - Gheyath K Nasrallah
- Biomedical Research Center, Qatar University, Doha 2713, Qatar.
- Biomedical Science Department, College of Health Sciences, Qatar University, Doha 2713, Qatar.
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6
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Gencay M, Seffner A, Pabinger S, Gautier J, Gohl P, Weizenegger M, Neofytos D, Batrla R, Woeste A, Kim HS, Westergaard G, Reinsch C, Brill E, Thuy PTT, Hoang BH, Sonderup M, Spearman CW, Brancaccio G, Fasano M, Gaeta GB, Santantonio T, Kaminski WE. Detection of in vivo hepatitis B virus surface antigen mutations-A comparison of four routine screening assays. J Viral Hepat 2018; 25:1132-1138. [PMID: 29660206 DOI: 10.1111/jvh.12915] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/04/2017] [Accepted: 03/25/2018] [Indexed: 12/12/2022]
Abstract
An important requirement for a state-of-the-art hepatitis B surface antigen (HBsAg) screening assay is reliable detection of mutated HBsAg. Currently, there is a striking shortage of data regarding the detection rates of in vivo HBsAg mutations for these clinically important assays. Therefore, we compared the detection rates of four commercial HBsAg screening assays using a global cohort of 1553 patients from four continents with known HBV genotypes. These samples, which represent the broadest spectrum of known and novel HBsAg major hydrophilic region (MHR) mutations to date, were analyzed for the presence of HBsAg using the Roche Elecsys® HBsAg II Qualitative, Siemens ADVIA Centaur XP HBsAg II, Abbott Architect HBsAg Qualitative II and DiaSorin Liaison® HBsAg Qualitative assays, respectively. Of the 1553 samples, 1391 samples could be sequenced; of these, 1013 (72.8%) carried at least one of the 345 currently known amino acid substitutions (distinct HBsAg mutation) in the HBsAg MHR. All 1553 patient samples were positive for HBsAg using the Elecsys® HBsAg II Qual assay, with a sensitivity (95% confidence interval) of 99.94% (99.64%-100%), followed by the Abbott Architect 99.81% (99.44%-99.96%), Siemens ADVIA 99.81% (99.44%-99.96%) and DiaSorin Liaison® 99.36% (98.82%-99.69%) assays, respectively. Our results indicate that the Elecsys® HBsAg II Qual assay exhibits the highest sensitivity among the commercial HBsAg screening assays, and demonstrate that its capacity to detect HBV infection is not compromised by HBsAg MHR mutants.
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Affiliation(s)
- M Gencay
- Roche Diagnostics International Ltd, Rotkreuz, Switzerland
| | - A Seffner
- Department of Molecular Genetics and Microbiology, MVZ Labor Dr. Limbach & Kollegen GbR, Heidelberg, Germany
| | - S Pabinger
- Health and Environment Department, Molecular Diagnostics, Austrian Institute of Technology, Vienna, Austria
| | - J Gautier
- Cerba Spécimen Services, Saint-Ouen l'Aumône, France
| | - P Gohl
- Bioscientia, Institute for Medical Diagnostics GmbH, Ingelheim, Germany
| | - M Weizenegger
- Department of Molecular Genetics and Microbiology, MVZ Labor Dr. Limbach & Kollegen GbR, Heidelberg, Germany
| | - D Neofytos
- Roche Diagnostics International Ltd, Rotkreuz, Switzerland
| | - R Batrla
- Roche Diagnostics International Ltd, Rotkreuz, Switzerland
| | - A Woeste
- Roche Diagnostics GmbH, Penzberg, Germany
| | - H S Kim
- Department of Laboratory Medicine, Yonsei University College of Medicine, Severance Hospital, Seoul, Korea
| | - G Westergaard
- Roche Diagnostics International Ltd, Rotkreuz, Switzerland
| | - C Reinsch
- Roche Diagnostics GmbH, Mannheim, Germany
| | - E Brill
- Bioscientia, Institute for Medical Diagnostics GmbH, Ingelheim, Germany
| | - P T T Thuy
- Hepatology Department, Medic Medical Center, Ho Chi Minh City, Vietnam
| | - B H Hoang
- Gastroenterology Department, Ho Chi Minh City University Medical Center, Ho Chi Minh City, Vietnam
| | - M Sonderup
- Division of Hepatology and Department of Medicine, University of Cape Town and Groote Schuur Hospital, Cape Town, South Africa
| | - C W Spearman
- Division of Hepatology and Department of Medicine, University of Cape Town and Groote Schuur Hospital, Cape Town, South Africa
| | - G Brancaccio
- Infectious Diseases and Viral Hepatitis Unit, University of Campania Luigi Vanvitelli, Naples, Italy
| | - M Fasano
- Infectious Diseases Unit, Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy
| | - G B Gaeta
- Infectious Diseases and Viral Hepatitis Unit, University of Campania Luigi Vanvitelli, Naples, Italy
| | - T Santantonio
- Infectious Diseases Unit, Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy
| | - W E Kaminski
- Bioscientia, Institute for Medical Diagnostics GmbH, Ingelheim, Germany
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Lou S, Taylor R, Pearce S, Kuhns M, Leary T. An ultra-sensitive Abbott ARCHITECT ® assay for the detection of hepatitis B virus surface antigen (HBsAg). J Clin Virol 2018; 105:18-25. [PMID: 29843004 DOI: 10.1016/j.jcv.2018.05.009] [Citation(s) in RCA: 31] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/02/2018] [Revised: 05/14/2018] [Accepted: 05/20/2018] [Indexed: 12/18/2022]
Abstract
BACKGROUND Critical to the identification of HBV infection and the prevention of transfusion transmitted disease is the sensitive and accurate detection of Hepatitis B virus surface antigen (HBsAg). Improvements in HBsAg assay sensitivity approaching the performance of nucleic acid testing (NAT) are essential to further reduce the detection window for acute HBV infection in regions where NAT is not widely available. OBJECTIVES AND STUDY DESIGN An improved HBsAg assay on the fully-automated Abbott ARCHITECT® platform was developed to improve sensitivity, mutant and genotype detection. RESULTS The analytical sensitivity of the improved prototype assay is 5.2 mIU/ml, which is 3.86- to 14.54-fold more sensitive than comparator assays based on the WHO International Reference Standard. The enhanced sensitivity was also demonstrated with 27 HBV seroconversion panels, detecting more panel members (191 of 364) vs. the ARCHITECT® Qual I (144), Qual II (160) and PRISM® (148) HBsAg assays. Further, the assay detected 7 of 12 HBV DNA positive/HBsAg negative samples, and detected all evaluated mutants and genotypes with higher sensitivity than the comparator assays. The improvement in sensitivity did not diminish assay specificity, attaining 100% (95% CI, 99.97-100%) on 10,633 blood donors. CONCLUSIONS An Abbott ARCHITECT® HBsAg assay with clinical performance approaching that of mini-pool NAT (approximately 100 copies/ml was developed. The assay has superior HBsAg mutant and genotype detection and specificity, all of which are important for the diagnosis and management of HBV infection.
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Affiliation(s)
- Sheng Lou
- Diagnostics Research, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL,60064, USA.
| | - Russell Taylor
- Diagnostics Research, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL,60064, USA.
| | - Sandra Pearce
- Diagnostics Research, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL,60064, USA.
| | - Mary Kuhns
- Diagnostics Research, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL,60064, USA.
| | - Thomas Leary
- Diagnostics Research, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL,60064, USA.
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Kim HS, Chen X, Xu M, Yan C, Liu Y, Deng H, Hoang BH, Thuy PTT, Wang T, Yan Y, Zeng Z, Gencay M, Westergaard G, Pabinger S, Kriegner A, Nauck M, Seffner A, Gohl P, Hübner K, Kaminski WE. Frequency of hepatitis B surface antigen variants (HBsAg) in hepatitis B virus genotype B and C infected East- and Southeast Asian patients: Detection by the Elecsys ® HBsAg II assay. J Clin Virol 2018; 103:48-56. [PMID: 29655170 DOI: 10.1016/j.jcv.2018.04.005] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/22/2018] [Revised: 03/29/2018] [Accepted: 04/04/2018] [Indexed: 02/06/2023]
Abstract
BACKGROUND To avoid false negative results, hepatitis B surface antigen (HBsAg) assays need to detect samples with mutations in the immunodominant 'a' determinant region, which vary by ethnographic region. OBJECTIVE We evaluated the prevalence and type of HBsAg mutations in a hepatitis B virus (HBV)-infected East- and Southeast Asian population, and the diagnostic performance of the Elecsys® HBsAg II Qualitative assay. STUDY DESIGN We analyzed 898 samples from patients with HBV infection from four sites (China [Beijing and Guangzhou], Korea and Vietnam). HBsAg mutations were detected and sequenced using highly sensitive ultra-deep sequencing and compared between the first (amino acids 124-137) and second (amino acids 139-147) loops of the 'a' determinant region using the Elecsys® HBsAg II Qualitative assay. RESULTS Overall, 237 distinct amino acid mutations in the major hydrophilic region were identified; mutations were present in 660 of 898 HBV-infected patient samples (73.5%). Within the pool of 237 distinct mutations, the majority of the amino acid mutations were found in HBV genotype C (64.8%). We identified 25 previously unknown distinct mutations, mostly prevalent in genotype C-infected Korean patients (n = 18) followed by Chinese (n = 12) patients. All 898 samples were correctly identified by the Elecsys® HBsAg II Qualitative assay. CONCLUSIONS We observed 237 distinct (including 25 novel) mutations, demonstrating the complexity of HBsAg variants in HBV-infected East- and Southeast Asian patients. The Elecsys® HBsAg II Qualitative assay can reliably detect HBV-positive samples and is suitable for routine diagnostic use in East and Southeast Asia.
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Affiliation(s)
- Hyon Suk Kim
- Department of Laboratory Medicine, Yonsei University College of Medicine, Severance Hospital, Seoul, South Korea
| | - Xinyue Chen
- Department of Liver Diseases, You'an Hospital, Capital Medical University, Beijing, China
| | - Min Xu
- Hepatology Department, Guangzhou No. 8 People's Hospital, Guangzhou, China
| | - Cunling Yan
- Department of Clinical Laboratory, Peking University First Hospital, Beijing, China
| | - Yali Liu
- Department of Liver Diseases, You'an Hospital, Capital Medical University, Beijing, China
| | - Haohui Deng
- Hepatology Department, Guangzhou No. 8 People's Hospital, Guangzhou, China
| | - Bui Huu Hoang
- Gastroenterology Department, Ho Chi Minh City University Medical Center, Ho Chi Minh City, Vietnam
| | - Pham Thi Thu Thuy
- Hepatology Department, Medic Medical Center, Ho Chi Minh City, Vietnam
| | | | - Yiwen Yan
- Roche Diagnostics Ltd., Shanghai, China
| | - Zhen Zeng
- Roche Diagnostics Ltd., Shanghai, China
| | - Mikael Gencay
- Roche Diagnostics International Ltd., Rotkreuz, Switzerland
| | | | - Stephan Pabinger
- AIT Austrian Institute of Technology, Molecular Diagnostics, Center for Health and Bioresources, Vienna, Austria
| | | | - Markus Nauck
- Bioscientia Institute for Medical Diagnostics GmbH, Ingelheim, Germany
| | - Anja Seffner
- MVZ Labor Dr. Limbach & Kollegen GbR, Department of Molecular Genetics and Microbiology, Heidelberg, Germany
| | - Peter Gohl
- Bioscientia Institute for Medical Diagnostics GmbH, Ingelheim, Germany
| | - Kirsten Hübner
- Bioscientia Institute for Medical Diagnostics GmbH, Ingelheim, Germany
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9
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Gencay M, Vermeulen M, Neofytos D, Westergaard G, Pabinger S, Kriegner A, Seffner A, Gohl P, Huebner K, Nauck M, Kaminski WE. Substantial variation in the hepatitis B surface antigen (HBsAg) in hepatitis B virus (HBV)-positive patients from South Africa: Reliable detection of HBV by the Elecsys HBsAg II assay. J Clin Virol 2018; 101:38-43. [DOI: 10.1016/j.jcv.2018.01.011] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/07/2017] [Revised: 01/12/2018] [Accepted: 01/21/2018] [Indexed: 02/07/2023]
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10
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Erdag B, Balcioglu KB, Bahadir AO, Hinc D, Ibrahimoglu O, Bahar A, Basalp A, Yucel F. Cloning of anti-HBsAg single-chain variable fragments from hybridoma cells for one-step ELISA. BIOTECHNOL BIOTEC EQ 2017. [DOI: 10.1080/13102818.2017.1348256] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/21/2023] Open
Affiliation(s)
- Berrin Erdag
- TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey
| | - Koray Bertan Balcioglu
- TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey
| | - Aylin Ozdemir Bahadir
- TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey
| | - Duygu Hinc
- TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey
| | - Ozlem Ibrahimoglu
- TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey
| | - Aydin Bahar
- TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey
| | - Aynur Basalp
- Sağlık Yüksekokulu, Mehmet Akif Ersoy Üniversitesi, Department of Health Managment, İstiklal Yerleşkesi, Burdur, Turkey
| | - Fatima Yucel
- TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey
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11
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Liang RL, Yang YS, Zhou JW, Liu TC, Xu XP, Liang QN, Chen ZH, Dong ZN, Wu YS. Dual-Labeled Time-Resolved Immunofluorometric Assay for the Simultaneous Quantitative Detection of Hepatitis B Virus Antigens in Human Serum. J Fluoresc 2016; 27:309-316. [PMID: 27878521 DOI: 10.1007/s10895-016-1959-0] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/21/2016] [Accepted: 10/19/2016] [Indexed: 12/27/2022]
Abstract
In this paper, a novel time-resolved fluorescence immunoassay (TRFIA) is described that allows the simultaneous quantitative detection of hepatitis B virus surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in human serum to aid the diagnosis and monitoring of hepatitis B virus infection. The proposed method was developed based on a two-step sandwich immunoassay protocol in which monoclonal antibodies against HBsAg and HBeAg were co-coated in 96 microtitration wells, then tracer polyclonal antibodies against HBsAg labeled with samarium and tracer monoclonal antibodies against HBeAg labeled with europium chelates were used for detection. The detection range was 0.1-150 IU/mL for HBsAg and 0.5-160 PEIU/mL for HBeAg, and the detection limits were 0.03 IU/L and 0.09 PEIU/ml, respectively. The intra- and inter-assay coefficients of variation were below 8 % for both virus antigens. The dilution linearity and accuracy of the assay were satisfactory. No statistically significant differences were observed in sensitivity or specificity for the serum samples between the dual-label TRFIA and a commercial single-label TRFIA. These results demonstrate that an effective, reliable and convenient HBsAg/HBeAg dual-label TRFIA was successfully developed that may be clinically applicable for blood screening to monitor the course of hepatitis B virus infection and predict treatment responses.
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Affiliation(s)
- Rong-Liang Liang
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Yun-Sen Yang
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Jian-Wei Zhou
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Tian-Cai Liu
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Xu-Ping Xu
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Qian-Ni Liang
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Zhen-Hua Chen
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Zhi-Ning Dong
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China
| | - Ying-Song Wu
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China.
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12
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Li J, Cheng B, Yang L, Zhao Y, Pan M, Zheng G, Xu X, Hu J, Xiao T, Cai Y. Development and Implementation of Autoverification Rules for ELISA Results of HBV Serological Markers. SLAS Technol 2016; 21:642-51. [PMID: 26311059 DOI: 10.1177/2211068215601612] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/24/2015] [Indexed: 02/05/2023]
Abstract
Autoverification is a process of using computer-based rules to verify clinical laboratory test results without manual review. But to date, there are few published articles on the use of autoverification over the course of years in a clinical laboratory. In our study, we firstly described the development and implementation of autoverification rules for enzyme-linked immunosorbent assay (ELISA) results of hepatitis B virus (HBV) serological markers in a clinical immunology laboratory. We designed the autoverification rules for HBV by using Boolean logic on five clinically used serological markers in accordance with the framework of AUTO-10A, issued by the American Clinical Laboratory Standards Institute in 2006. The rules were written into the laboratory information system (LIS) and installed in the computer, so we could use the LIS to screen the test results. If the results passed the autoverification rules, they could be sent to doctors immediately. To evaluate the autoverification rules, we applied the real-time data of 11,585 patients with the autoverification rules. The autoverification rate of the five HBV serological markers was 79.5%. Furthermore, the turnaround time (TAT) was reduced by 38% (78 minutes vs. 126 minutes). The error rate was nearly eliminated. These results show that using LIS with autoverification rules can shorten TAT, enhance efficiency, and reduce manual review errors.
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Affiliation(s)
- Jiancheng Li
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Bizhen Cheng
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Li Yang
- Department of Clinical Laboratory, Shantou Central Hospital, Guangdong, People's Republic of China
| | - Ying Zhao
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Meichen Pan
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Gaozhe Zheng
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Xiaoyan Xu
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Jing Hu
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Tongtong Xiao
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
| | - Yingmu Cai
- Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Guangdong, People's Republic of China
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13
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De Paschale M, Manco MT, Belvisi L, Cagnin D, Cerulli T, Paganini A, Arpino O, Cianflone A, Agrappi C, Mirri P, Clerici P. Evaluation of LIAISON® XL system for HBsAg, and anti-HCV and anti-HIV/Ag p24. J Med Virol 2016; 89:489-496. [PMID: 27467710 DOI: 10.1002/jmv.24648] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 07/18/2016] [Indexed: 12/17/2022]
Abstract
The aim of this study was to compare the data obtained using the new LIAISON® XL chemiluminescence system to search for HBsAg, anti-HCV, and anti-HIV1-2/p24 Ag with those obtained using the VITROS system currently adopted by the Microbiology Unit of the Hospital of Legnano. Routine samples of patients who were referred by practitioners for the determination of HBsAg (1,000 samples) and/or anti-HCV (1,002 samples) and/or anti-HIV1-2 (995 samples) were simultaneously analyzed using both systems. The concordant positive and discordant samples were re-examined for confirmation by means of an HBsAg neutralization assay, anti-HCV immunoblot, or anti-HIV1-2 Western blot; HBV-DNA, or HCV-RNA or HIV-RNA was also sought in the discordant samples. Samples of patients known to be positive were tested (100 HBsAg positive, 100 anti-HCV positive, and 100 HIV 1-2 positive) as well throughout treatment, with viremia levels becoming undetectable after treatment. The HBsAg, anti-HCV, and anti-HIV1-2 concordance between the two systems in routine series was respectively 99.8%, 98.5% and 99.7%, and 100% for all markers in samples known positive. The various molecular biology and confirmatory tests of the discordant samples were all negative (except for one anti-HCV positive sample). Measure of Cohen's kappa coefficient for HBsAg, anti-HCV, and anti-HIV gave K values of respectively 0.992, 0.946, and 0.980. In conclusion, the performance of the LIAISON® XL system in the routine laboratory determination for all three markers was comparable with that of the VITROS system. J. Med. Virol. 89:489-496, 2017. © 2016 Wiley Periodicals, Inc.
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Affiliation(s)
| | - Maria Teresa Manco
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Luisa Belvisi
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Debora Cagnin
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Teresa Cerulli
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Alessia Paganini
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Olivia Arpino
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Annalisa Cianflone
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Carlo Agrappi
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Paola Mirri
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
| | - Pierangelo Clerici
- Microbiology Unit, ASST Ovest Milanese, Hospital of Legnano, Milan, Italy
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14
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Osiowy C, Kowalec K, Giles E. Discordant diagnostic results due to a hepatitis B virus T123A HBsAg mutant. Diagn Microbiol Infect Dis 2016; 85:328-333. [DOI: 10.1016/j.diagmicrobio.2016.04.004] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/25/2016] [Revised: 04/04/2016] [Accepted: 04/06/2016] [Indexed: 12/20/2022]
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15
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Ye Q, Shang SQ, Li W. A new vaccine escape mutant of hepatitis B virus causes occult infection. Hum Vaccin Immunother 2015; 11:407-10. [PMID: 25692622 DOI: 10.4161/21645515.2014.994461] [Citation(s) in RCA: 20] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022] Open
Abstract
There is growing public concern regarding assay sensitivity to HBsAg mutants in clinical diagnosis and vaccine escape. The aim of this study is to introduce a new HBsAg mutant strain. The serum samples were those of patient X at the age of 3 months and 3 years respectively, and of her mother immediately before parturition, which were used to amplify the HBsAg-coding DNA fragments by PCR. The HBsAg DNA sequences were translated into their corresponding amino acid sequences and then aligned in pubmed with nucleotide blast. The sequencing data of S coding regions shows that patient X has been infected by a new HBV variant with an A to C substitution at nt431, resulting in an Asp(GAC)to Ala(GCC) substitution at aa144 of major protein; CC to AA substitution at nt359 and nt360, resulting in an Pro(CCC) to Gln(CAA) substitution at aa120 of pre "a" epitope; A to G substitution at nt491, resulting in an Glu(GAG) to Gly(GGG) substitution at aa164 of post "a" epitope. Three new mutations (S171F, S174N and Q181R) at the antigenic epitopes of HBV presented by HLA class I molecules are found. The HBV mutant strain causes vaccine escape and occult infection.
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Key Words
- ALP, alkaline phosphatase
- ALT, alanine aminotransferase
- AST, aspartate aminotransferase
- AchE, acetylcholin esterase
- Alb, albumin
- Anti-HBc, antibodies to hepatitis B core antigen
- Anti-HBs, antibodies to HBsAg
- CMV, cytomegalovirus
- CRP, C-reactive protein
- EBNA, Epstein-Barr nuclear antigen
- EBV, Epstein-Barr virus
- EIA, enzyme immunoassay
- HBV
- HBsAg, hepatitis B surface antigen
- HCV, hepatitis C virus
- Hb, hemoglobin
- LDH, lactate dehydrogenase
- Plt, platelet count
- RBC, red blood cell count
- T-Bil, total bilirubin
- TG, triglycerides
- TP, total protein
- UA, uric acid
- VCA, viral capsid antigen
- WBC, white blood cell count
- anti-HA, antibody to hepatitis A
- gene mutation
- occult infection
- vaccine escape
- γGTP, γ-glutamyl transferase
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Affiliation(s)
- Qing Ye
- a Clinical Laboratory; The Children's Hospital; School of Medicine ; Zhejiang University ; Hangzhou , PR China
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16
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Liu YP, Yao CY. Rapid and quantitative detection of hepatitis B virus. World J Gastroenterol 2015; 21:11954-11963. [PMID: 26576084 PMCID: PMC4641117 DOI: 10.3748/wjg.v21.i42.11954] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/23/2015] [Revised: 07/29/2015] [Accepted: 09/14/2015] [Indexed: 02/06/2023] Open
Abstract
Despite availability of a universal vaccine, hepatitis B virus (HBV) infection has a huge impact on public health worldwide. Accurate and timely diagnosis of HBV infection is needed. Rapid developments have been made in the diagnostic and monitoring methods for HBV infection, including serological and molecular assays. In clinical practice, qualitative hepatitis B surface antigen (HBsAg) testing has long served as a diagnostic marker for individuals infected with HBV. More recently, HBsAg level has been used to predict treatment outcome when determined early during treatment or at baseline. However, identification of HBV DNA positive cases that do not have detectable HBsAg has encouraged the application of molecular tests. Hence, combination of quantitative detection of HBV DNA and HBsAg can be used to discriminate patients during the course of HBV infection and to monitor therapy. This article reviews the most commonly used quantitative methods for HBsAg and HBV DNA.
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17
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Hirzel C, Pfister S, Gorgievski-Hrisoho M, Wandeler G, Zuercher S. Performance of HBsAg point-of-care tests for detection of diagnostic escape-variants in clinical samples. J Clin Virol 2015. [PMID: 26209374 DOI: 10.1016/j.jcv.2015.05.024] [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] [Indexed: 01/01/2023]
Abstract
BACKGROUND Hepatitis B viruses (HBV) harboring mutations in the a-determinant of the Hepatitis B surface antigen (HBsAg) are associated with reduced reactivity of HBsAg assays. OBJECTIVES To evaluate the sensitivity and specificity of three HBsAg point-of-care tests for the detection of HBsAg of viruses harboring HBsAg mutations. STUDY DESIGN A selection of 50 clinical plasma samples containing HBV with HBsAg mutations was used to evaluate the performance of three HBsAg point-of-care tests (Vikia(®), bioMérieux, Marcy-L'Étoile, France. Alere Determine HBsAg™, Iverness Biomedical Innovations, Köln, Germany. Quick Profile™, LumiQuick Diagnostics, California, USA) and compared to the ARCHITECT HBsAg Qualitative(®) assay (Abbott Laboratories, Sligo, Ireland). RESULTS The sensitivity of the point-of-care tests ranged from 98% to 100%. The only false-negative result occurred using the Quick Profile™ assay with a virus harboring a D144A mutation. CONCLUSIONS The evaluated point-of-care tests revealed an excellent sensitivity in detecting HBV samples harboring HBsAg mutations.
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Affiliation(s)
- Cédric Hirzel
- Department of Infectious Diseases, Bern University Hospital and University of Bern, Switzerland.
| | - Stefan Pfister
- Institute for Infectious Diseases, University of Bern, Bern, Switzerland
| | | | - Gilles Wandeler
- Department of Infectious Diseases, Bern University Hospital and University of Bern, Switzerland; Institute of Social and Preventive Medicine, University of Bern, Bern, Switzerland
| | - Samuel Zuercher
- Institute for Infectious Diseases, University of Bern, Bern, Switzerland
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18
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Schmidt M, Jimenez A, Mühlbacher A, Oota S, Blanco L, Sakuldamrongpanich T, Schennach H, Seifried E. Head-to-head comparison between two screening systems for HBsAG, anti-HBc, anti-HCV and HIV combination immunoassays in an international, multicentre evaluation study. Vox Sang 2015; 109:114-21. [PMID: 25899479 DOI: 10.1111/vox.12259] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2014] [Revised: 01/19/2015] [Accepted: 01/19/2015] [Indexed: 01/05/2023]
Abstract
BACKGROUND Mandatory screening of blood donations for hepatitis B and hepatitis C viruses and human immunodeficiency viruses 1 and 2 requires assays with exceptional sensitivity and specificity. This study reports the results from a direct head-to-head comparison of the Elecsys HBsAG II, Elecsys Anti-HBc, Elecsys Anti-HCV II and Elecsys HIV combi PT immunoassays with the respective ABBOTT PRISM/Architect instrument immunoassays in a multicentre blood bank evaluation study. STUDY DESIGN AND METHODS Assay validation was performed in the blood screening laboratories of four blood bank centres in Austria, Germany, Spain and Thailand, where both first-time donor samples (approximately 6000 donors) and repeat donor samples (approximately 14,000 donors) were screened. RESULTS Of all screened donor samples, 93 (0.46%) were confirmed to be positive using assays from both manufacturers. The specificity of all immunoassays was >99.5% and was comparable between first-time and multiple-time donors. A direct comparison between the assays from Roche and ABBOTT according to Bland and Altman analysis demonstrated equivalent quality. CONCLUSIONS These results suggest that the Elecsys immunoassays for HBV, HCV and HIV infection, with a comparative sensitivity of 100% and a specificity exceeding the common technical specification threshold of >99.5%, meet the stringent performance criteria stipulated for blood donor screening for these infectious agents. Significant differences in the specificity between first-time and repeat donors were not detectable.
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Affiliation(s)
- M Schmidt
- DRK-Blutspendedienst Baden-Württemberg, Hessen, Institut für Transfusionsmedizin und Immunhämatologie, Frankfurt, Germany
| | - A Jimenez
- Centro de Hemoterapia y Hemodonación Castilla y León, Valladolid, Spain
| | - A Mühlbacher
- Zentralinstitut fuer Bluttransfusion und Immunologie Abteilung der Tilak Universitätsklinik LKH Innsbruck, Innsbruck, Austria
| | - S Oota
- National Blood Centre, Thai Red Cross Society, Bangkok, Thailand
| | - L Blanco
- Centro de Hemoterapia y Hemodonación Castilla y León, Valladolid, Spain
| | | | - H Schennach
- Zentralinstitut fuer Bluttransfusion und Immunologie Abteilung der Tilak Universitätsklinik LKH Innsbruck, Innsbruck, Austria
| | - E Seifried
- DRK-Blutspendedienst Baden-Württemberg, Hessen, Institut für Transfusionsmedizin und Immunhämatologie, Frankfurt, Germany
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Thibault V, Laperche S, Thiers V, Sayon S, Letort MJ, Delarocque-Astagneau E, Antona D. Molecular epidemiology and clinical characteristics of hepatitis B identified through the French mandatory notification system. PLoS One 2013; 8:e75267. [PMID: 24086488 PMCID: PMC3783366 DOI: 10.1371/journal.pone.0075267] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2013] [Accepted: 08/14/2013] [Indexed: 02/07/2023] Open
Abstract
BACKGROUND & AIMS Strains responsible for acute hepatitis B infections (AHB) in France have not been characterized. This study was first designed to analyze the molecular epidemiology of AHB and second to describe the differences between AHB and chronic hepatitis B (CHB) exacerbations. METHODS This prospective study was based on the French mandatory notification system for AHB. 147 samples corresponding to declared cases were shipped to a central laboratory for classification as AHB or CHB according to the level of anti-HBc IgM and anti-HBc avidity. RESULTS Based on biological marker values and file examination, 75 cases (59%) were classified as AHB. Independently of the acute or chronic status, genotype A (57%), D (22%) and E (14%) were the most prevalent and no phylogenetic clustering was observed among HBV sequences (n=68). Precore or basal core-promoter variants were not particularly associated with disease severity but were more prevalent in CHB. No antiviral resistant strains or immune-escape HBsAg was observed. HBV viral loads in AHB or CHB were comparable but with opposite distributions. ALT levels reached 10 times the upper normal value in 94% of AHB but only in 24% of CHB. CONCLUSIONS After rigorous classification, no major difference at the genetic level was found between HBV strains isolated from AHB and CHB. Absence of potentially deleterious variant detection is reassuring. When based upon HBsAg and anti-HBc IgM determination, AHB notification may falsely include more than 40% CHB, leading to an important risk of bias in national surveillance programs of AHB.
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Affiliation(s)
- Vincent Thibault
- Virology Laboratory, Hôpital Pitié-Salpêtrière, Assistance Publique (APHP), and Pierre et Marie Curie University, Paris, France
- * E-mail:
| | - Syria Laperche
- National reference center for hepatitis B and C in blood transfusion, National Institute of blood transfusion, Paris, France
| | | | - Sophie Sayon
- Virology Laboratory, Hôpital Pitié-Salpêtrière, Assistance Publique (APHP), and Pierre et Marie Curie University, Paris, France
| | - Marie-José Letort
- Infectious Diseases Department, National Institute for Public Health Surveillance (Institut de veille sanitaire), Saint-Maurice, France
| | - Elisabeth Delarocque-Astagneau
- Emerging Diseases Epidemiology Unit and Pharmacoepidemiology and Infectious Diseases Unit, Institut Pasteur, Paris, France
| | - Denise Antona
- Infectious Diseases Department, National Institute for Public Health Surveillance (Institut de veille sanitaire), Saint-Maurice, France
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Cabezas-Fernandez MT, Cabeza-Barrera MI. Introduction of an automated system for the diagnosis and quantification of hepatitis B and hepatitis C viruses. Open Virol J 2012; 6:122-34. [PMID: 23284598 PMCID: PMC3531716 DOI: 10.2174/1874357901206010122] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/09/2012] [Revised: 09/18/2012] [Accepted: 09/20/2012] [Indexed: 02/07/2023] Open
Abstract
Hepatitis B virus (HBV) and Hepatitis C virus (HCV) infections pose major public health problems because of their prevalence worldwide. Consequently, screening for these infections is an important part of routine laboratory activity. Serological and molecular markers are key elements in diagnosis, prognosis and treatment monitoring for HBV and HCV infections. Today, automated chemiluminescence immunoassay (CLIA) analyzers are widely used for virological diagnosis, particularly in high-volume clinical laboratories. Molecular biology techniques are routinely used to detect and quantify viral genomes as well as to analyze their sequence; in order to determine their genotype and detect resistance to antiviral drugs. Real-time PCR, which provides high sensitivity and a broad dynamic range, has gradually replaced other signal and target amplification technologies for the quantification and detection of nucleic acid. The next-generation DNA sequencing techniques are still restricted to research laboratories.The serological and molecular marker methods available for HBV and HCV are discussed in this article, along with their utility and limitations for use in Chronic Hepatitis B (CHB) diagnosis and monitoring.
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21
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Analysis of mutations in the S gene of hepatitis B virus strains in patients with chronic infection by online bioinformatics tools. J Clin Microbiol 2012; 51:163-8. [PMID: 23115258 DOI: 10.1128/jcm.01630-12] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
Hepatitis B virus (HBV) genomes show a high rate of mutations. This can lead to a variety of amino acid changes in the surface and polymerase genes, causing changes in viral protein conformation that can result in diminished antibody binding or decreased secretion of surface antigen (HBsAg). HBV monitoring increasingly relies on HBsAg detection and quantification, and therefore epidemiological data on HBsAg mutations are needed. We therefore analyzed the frequency of HBsAg mutations possibly influencing the quantification of HBsAg (MUPIQHs) in an unselected patient collective. To this end, we determined the HBV surface and polymerase gene sequences of an unselected patient collective of 237 individuals chronically infected with HBV and analyzed the MUPIQHs in these sequences using three different online HBV sequence analysis tools. We found that 17 or 34% of the patients, depending on the online interpretation algorithm used, harbored MUPIQHs and that MUPIQHs were not significantly associated with the duration of disease, treatment, or HBV genotype. Thus, this study shows that a substantial amount of HBV sequences derived from unselected patients chronically infected with HBV carry MUPIQHs, and therefore the reliability of routine quantitative and qualitative HBsAg tests needs to be reevaluated.
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Alavian SM, Carman WF, Jazayeri SM. HBsAg variants: diagnostic-escape and diagnostic dilemma. J Clin Virol 2012; 57:201-8. [PMID: 22789139 DOI: 10.1016/j.jcv.2012.04.027] [Citation(s) in RCA: 59] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2011] [Revised: 01/23/2012] [Accepted: 04/18/2012] [Indexed: 12/11/2022]
Abstract
A wide variety of commercial assays is available for the detection of hepatitis B surface antigen (HBsAg). Clearly, the sensitivity of an assay to detect a variant is dependent on the anti-HBs usage. Thus, it is not surprising that there are examples of variants that cannot be detected by all assays. Data from Europe, Asia and Africa about HBsAg variants which are not recognized by either monoclonal or polyclonal antibodies specific for wild-type group 'a' determinant, but positive by DNA polymerase chain reaction (PCR) in chronic patients and from vaccinated children are increasing. This would impose a challenge for public health issues of hepatitis B virus. In this review we tried to summarize the discrepancies between results of HBsAg assays and to explain some rationales for these inconsistencies.
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Affiliation(s)
- Seyed Moayed Alavian
- Baqiyatallah University of Medical Sciences, Baqiyatallah Research Centre for Gastroenterology and Liver Disease, Tehran, Iran
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Verheyen J, Neumann-Fraune M, Berg T, Kaiser R, Obermeier M. The detection of HBsAg mutants expressed in vitro using two different quantitative HBsAg assays. J Clin Virol 2012; 54:279-81. [DOI: 10.1016/j.jcv.2012.04.010] [Citation(s) in RCA: 28] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/02/2012] [Revised: 04/04/2012] [Accepted: 04/17/2012] [Indexed: 10/28/2022]
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Servant-Delmas A, Mercier-Darty M, Ly TD, Wind F, Alloui C, Sureau C, Laperche S. Variable capacity of 13 hepatitis B virus surface antigen assays for the detection of HBsAg mutants in blood samples. J Clin Virol 2012; 53:338-45. [DOI: 10.1016/j.jcv.2012.01.003] [Citation(s) in RCA: 36] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/03/2012] [Accepted: 01/05/2012] [Indexed: 12/11/2022]
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False-negative hepatitis B virus (HBV) surface antigen in a vaccinated dialysis patient with a high level of HBV DNA in the United States. CLINICAL AND VACCINE IMMUNOLOGY : CVI 2012; 19:820-2. [PMID: 22441395 DOI: 10.1128/cvi.05696-11] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/27/2022]
Abstract
Screening with hepatitis B surface antigen (HBsAg) is highly recommended for at-risk individuals. Mutations in the HBsAg can result in an inability to detect the virus during routine screening. We describe a hemodialysis patient found to have high levels of hepatitis B virus (HBV) DNA and HBV antibody but negative HBsAg on two routine assays.
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Seto WK, Tanaka Y, Wong DKH, Lai CL, Shinkai N, Yuen JCH, Tong T, Fung J, Hung IFN, Yuen MF. Evidence of serologic activity in chronic hepatitis B after surface antigen (HBsAg) seroclearance documented by conventional HBsAg assay. Hepatol Int 2012; 7:98-105. [PMID: 24014110 PMCID: PMC3758508 DOI: 10.1007/s12072-012-9354-7] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/04/2011] [Accepted: 02/21/2012] [Indexed: 02/07/2023]
Abstract
BACKGROUND Possible serologic activity after hepatitis B surface antigen (HBsAg) seroclearance documented by conventional assays in chronic hepatitis B (CHB) has not been thoroughly investigated. METHODS We determined the levels of serum hepatitis B virus (HBV) DNA, hepatitis B core-related antigen (HBcrAg), and linearized HBsAg (CLEIA prototype) in 329 CHB patients (72.0% male) after HBsAg seroclearance was documented by a conventional HBsAg assay. RESULTS The median interval between presentation and HBsAg seroclearance was 69.4 months. The median age at HBsAg seroclearance was 50 years. Assays for serum HBV DNA, HBcrAg, and linearized HBsAg were performed at a median time interval of 11.2 months after HBsAg loss. Linearized HBsAg and HBcrAg were detectable in 85 (25.8%) and 69 (21%) patients, respectively, and one or both serologic markers were detectable in 133 patients (40.4%). Serum HBV DNA was detectable in only 7 patients (2.1%). There was no correlation between linearized HBsAg and HBcrAg levels (r = 0.095, p = 0.924). The incidences of detectable linearized HBsAg and HBcrAg did not differ between patient samples taken at 6-12 and >12 months after HBsAg seroclearance (p = 0.146 and 0.079, respectively). Among patients with detectable serologic markers, median levels of linearized HBsAg (p = 0.581) and HBcrAg (p = 0.951) did not significantly change with time after HBsAg seroclearance. CONCLUSION Using novel HBcrAg and linearized HBsAg assays, viral serologic activity after HBsAg seroclearance was demonstrated in more than 40% of CHB patients. These tests have potential applications in diagnosing and prognosticating CHB patients with HBsAg seroclearance.
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Affiliation(s)
- Wai-Kay Seto
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
| | - Yasuhito Tanaka
- />Department of Virology and Liver Unit, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan
| | - Danny Ka-Ho Wong
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
| | - Ching-Lung Lai
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
- />State Key Laboratory for Liver Research, Queen Mary Hospital, The University of Hong Kong, Hong Kong, China
| | - Noboru Shinkai
- />Department of Virology and Liver Unit, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan
| | - John Chi-Hang Yuen
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
| | - Teresa Tong
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
| | - James Fung
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
| | - Ivan Fan-Ngai Hung
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
| | - Man-Fung Yuen
- />Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Hong Kong, China
- />State Key Laboratory for Liver Research, Queen Mary Hospital, The University of Hong Kong, Hong Kong, China
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Development of an economic and efficient strategy to detect HBsAg: Application of “gray-zones” in ELISA and combined use of several detection assays. Clin Chim Acta 2011; 412:2046-51. [DOI: 10.1016/j.cca.2011.01.020] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/26/2010] [Revised: 01/03/2011] [Accepted: 01/18/2011] [Indexed: 11/23/2022]
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Farnik H, Lange CM, Hofmann WP, Berger A, Allwinn R, Welker MW, Trojan J, Sarrazin C, Herrmann E, Zeuzem S, Kronenberger B. Nucleos(t)ide analogue treatment reduces apoptotic activity in patients with chronic hepatitis B. J Clin Virol 2011; 52:204-9. [PMID: 21903459 DOI: 10.1016/j.jcv.2011.08.009] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2011] [Revised: 06/23/2011] [Accepted: 08/08/2011] [Indexed: 01/05/2023]
Abstract
BACKGROUND Reduction of necroinflammatory activity is a major goal of antiviral therapy of patients with chronic hepatitis B. Serum ALT does not detect all forms of cell death. OBJECTIVES To analyze dynamics of novel serum cell death markers for apoptosis and necrosis in association with virologic response to nucleos(t)ide (Nuc) analogue treatment. STUDY DESIGN Quantification of the M30-apoptosis neoepitope and the cytokeratin-18 (M65-necrosis) serum levels before and during treatment of patients with chronic hepatitis B with Nuc (n = 26). RESULTS Before treatment, M30-apoptotic activity was significantly correlated with M65-necrosis and fibrosis but not with serum ALT. During therapy with Nucs, cell death parameters M30-apoptosis, M65-necrosis, and ALT declined in association with virologic response. The most frequent cell death pattern was simultaneous decline of ALT and M30-apoptosis which occurred more frequently in patients with HBs-Antigen decline than in patients with HBs-Antigen increase during treatment (87.5% vs. 40.0%; p = 0.024). ALT decline in association with increase of M30 apoptosis was frequent in patients with HBs-Antigen increase during treatment (36.3%) but was not observed in patients with HBs-Antigen decline during treatment. CONCLUSION Decline of cell death parameters in association with decline of HBV-DNA and HBs-Antigen indicates a reduction in overall cell death activity during Nuc treatment supporting the concept that response to Nuc therapy reduces necroinflammatory activity and progression of liver disease.
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Affiliation(s)
- Harald Farnik
- Medizinische Klinik 1, Klinikum der Johann Wolfgang Goethe-Universität Frankfurt, Theodor-Stern-Kai 7, Haus 11, 60590 Frankfurt, Germany
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Sonneveld MJ, Rijckborst V, Boucher CA, Zwang L, Beersma MF, Hansen BE, Janssen HL. A comparison of two assays for quantification of Hepatitis B surface Antigen in patients with chronic hepatitis B. J Clin Virol 2011; 51:175-8. [DOI: 10.1016/j.jcv.2011.04.005] [Citation(s) in RCA: 43] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2010] [Revised: 04/07/2011] [Accepted: 04/11/2011] [Indexed: 02/07/2023]
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HBsAg blood screening and diagnosis: performance evaluation of the ARCHITECT HBsAg qualitative and ARCHITECT HBsAg qualitative confirmatory assays. Diagn Microbiol Infect Dis 2011; 70:479-85. [PMID: 21658874 DOI: 10.1016/j.diagmicrobio.2011.03.022] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2011] [Revised: 03/21/2011] [Accepted: 03/28/2011] [Indexed: 12/28/2022]
Abstract
A low initial reactive rate for screening assays is important for time- and cost-effective infectious disease testing. Therefore, the new ARCHITECT HBsAg Qualitative screening assay, in conjunction with the new ARCHITECT HBsAg Qualitative Confirmatory assay, was introduced. As the role of hepatitis B surface antigen (HBsAg) as surrogate marker for HBV resolution and the monitoring of drug effectiveness are becoming increasingly important, the established ARCHITECT HBsAg Quantitative assay remains available on the market. Precision, sensitivity, and specificity of the newly developed screening assay were in the range of established HBsAg assays. Seroconversion sensitivity was slightly superior compared to other commercially available assays. An initial reactive rate of 0.2% (without HBsAg-confirmed positive samples of 0.17%) for the ARCHITECT HBsAg Qualitative assay was observed. As the new screening assay is a 1-step assay format, the "high-dose hook effect" was investigated to assess the risk of false-negative results, but even very high positive HBsAg samples obtained signals clearly above the cutoff.
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Wursthorn K, Jaroszewicz J, Zacher BJ, Darnedde M, Raupach R, Mederacke I, Cornberg M, Manns MP, Wedemeyer H. Correlation between the Elecsys HBsAg II assay and the Architect assay for the quantification of hepatitis B surface antigen (HBsAg) in the serum. J Clin Virol 2011; 50:292-6. [DOI: 10.1016/j.jcv.2010.12.008] [Citation(s) in RCA: 58] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/23/2010] [Revised: 12/08/2010] [Accepted: 12/21/2010] [Indexed: 12/13/2022]
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An improved Abbott ARCHITECT assay for the detection of hepatitis B virus surface antigen (HBsAg). J Clin Virol 2011; 51:59-63. [PMID: 21367654 DOI: 10.1016/j.jcv.2011.01.019] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2010] [Revised: 01/17/2011] [Accepted: 01/19/2011] [Indexed: 02/07/2023]
Abstract
BACKGROUND The sensitive and accurate detection of hepatitis B virus surface antigen (HBsAg) is critical to the identification of infection and the prevention of transfusion transmitted disease. Improvement in HBsAg assay sensitivity is essential to reduce the window to detect an acute HBV infection. Additionally, the sensitive detection of HBsAg mutants that continue to evolve due to vaccine escape, immune selection and an error prone reverse transcriptase is a necessity. OBJECTIVES AND STUDY DESIGN A fully automated HBsAg prototype assay on the Abbott ARCHITECT instrument was developed to improve sensitivity and mutant detection. This magnetic microparticle-based assay utilizes anti-HBsAg monoclonal antibodies to capture antigen present in serum or plasma. Captured antigen is then detected using anti-HBsAg antibody conjugated with the chemiluminescent compound, acridinium. RESULTS The sensitivity of the ARCHITECT HBsAg prototype assay was improved as compared to the current ARCHITECT, PRISM, and competitor HBsAg assays. The enhancement in assay sensitivity was demonstrated by the use of commercially available HBV seroconversion panels. The prototype assay detected more panel members (185 of 383) vs. the current ARCHITECT (171), PRISM (181), or competitor HBsAg assays (73/140 vs. 62/140, respectively). The ARCHITECT prototype assay also efficiently detected all mutants evaluated. Finally, the sensitivity improvement did not compromise the specificity of the assay (99.94%). CONCLUSIONS An improved Abbott ARCHITECT HBsAg prototype assay with enhanced detection of HBsAg and HBsAg mutants, as well as equivalent specificity was developed for the detection, diagnosis, and management of HBV infection.
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Lee JM, Ahn SH. Quantification of HBsAg: Basic virology for clinical practice. World J Gastroenterol 2011; 17:283-9. [PMID: 21253386 PMCID: PMC3022287 DOI: 10.3748/wjg.v17.i3.283] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/22/2010] [Revised: 03/23/2010] [Accepted: 03/30/2010] [Indexed: 02/06/2023] Open
Abstract
Hepatitis B surface antigen (HBsAg) is produced and secreted through a complex mechanism that is still not fully understood. In clinical fields, HBsAg has long served as a qualitative diagnostic marker for hepatitis B virus infection. Notably, advances have been made in the development of quantitative HBsAg assays, which have allowed viral replication monitoring, and there is an opportunity to make maximal use of quantitative HBsAg to elucidate its role in clinical fields. Yet, it needs to be underscored that a further understanding of HBsAg, not only from clinical point of view but also from a virologic point of view, would enable us to deepen our insights, so that we could more widely expand and apply its utility. It is also important to be familiar with HBsAg variants and their clinical consequences in terms of immune escape mutants, issues resulting from overlap with corresponding mutation in the P gene, and detection problems for the HBsAg variants. In this article, we review current concepts and issues on the quantification of HBsAg titers with respect to their biologic nature, method principles, and clinically relevant topics.
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Avellón A, Echevarría JM, Weber B, Weik M, Schobel U, Willems WR, Gerlich WH. European collaborative evaluation of the enzygnost HBsAg 6.0 assay: Performance on hepatitis B virus surface antigen variants. J Med Virol 2010; 83:95-100. [DOI: 10.1002/jmv.21943] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
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Yang J, Kim JH, Kim Y. [Comparison of nine different qualitative HBsAg assay kits]. Korean J Lab Med 2010; 30:178-84. [PMID: 20445337 DOI: 10.3343/kjlm.2010.30.2.178] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/19/2022] Open
Abstract
BACKGROUND Qualitative hepatitis B surface antigen (HBsAg) assay kits are still commonly used in Korea where hepatitis B virus (HBV) infection is endemic. The accurate determination of HBsAg plays a crucial role in the diagnosis and prevention of HBV infection, especially in endemic areas. The aim of this study was to compare the detection sensitivities of 9 qualitative HBsAg assay kits. METHODS Seven pooled sera with HBsAg concentration ranging from 0.14 IU/mL to 29.96 IU/mL were prepared. The HBsAg concentration of each pooled serum was determined by a quantitative HBsAg assay, Architect HBsAg (Abbott Laboratories, Ireland). The fully automated immunoassay kits included Elecsys HBsAg (Roche Diagnostics, Germany) and Immulite 2000 HBsAg (DPC, USA) and the rapid tests included 5 immunochromatographic assay (ICA) kits and 2 reverse passive hemagglutination assay (RPHA) kits. RESULTS Elecsys HBsAg (Roche Diagnostics) showed positive result in pooled serum with HBsAg concentration of 0.14 IU/mL, but Immulite 2000 HBsAg (DPC) showed negative result in the same concentration. Although ICA kits showed variable results among different assay kits, all of them showed negative results in pooled sera with HBsAg concentration of < or = 1.89 IU/mL. Two RPHA kits showed negative results in pooled sera with HBsAg concentration of < or = 7.98 IU/mL. CONCLUSIONS Although ICAs were more sensitive than RPHAs, they had variable sensitivities for HBsAg and were less sensitive than the automated immunoassay kits. Therefore, ICAs and RPHAs should be used with caution in the screening tests for HBsAg and their sensitivities need to be improved.
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Affiliation(s)
- Jinyoung Yang
- Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea
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Detection of highly prevalent hepatitis B virus coinfection among HIV-seropositive persons in Ghana. J Clin Microbiol 2010; 48:3223-30. [PMID: 20631103 DOI: 10.1128/jcm.02231-09] [Citation(s) in RCA: 79] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
Simple hepatitis B surface antigen (HBsAg) tests may facilitate ascertainment of hepatitis B virus (HBV) infection in settings with high endemicity but limited infrastructure. We evaluated two rapid HBsAg tests and characterized HBV coinfection in a Ghanaian HIV-positive cohort. Samples from 838 patients were tested by the rapid assays Determine and Vikia and the reference assays Architect, Murex version 3, and Liaison Ultra. The assays were also evaluated using the 2nd International Standard, a seroconversion panel, and two mutant panels. HBsAg-positive samples underwent HBV DNA quantification by real-time PCR and surface and polymerase gene population sequencing. Overall, 140/838 patients (16.7%; 95% confidence interval, 14.2 to 19.2%) were HBsAg positive, and of these, 103/140 (73.6%) were e-antigen negative and 118/140 (84.3%) showed an HBV DNA level of >14 IU/ml (median, 8,279 IU/ml). Assay sensitivities and specificities were as follows: Architect, 97.9 and 99.6%; Liaison, 97.1 and 99.4%; Murex, 98.6 and 99.3%; Determine, 69.3 and 100%; and Vikia, 70.7 and 100%. With Determine, the limit of detection was >1.5 to 3.4 HBsAg IU/ml, and the median HBV DNA loads were 598 and 10,905 IU/ml in Determine-negative and -positive samples, respectively (P = 0.0005). Results were similar with the Vikia assay. HBV DNA sequencing indicated infection with genotype E in 82/86 (95.3%) patients. HBsAg mutations affected assay performance, including a T123A mutant that escaped detection by Architect. Major drug resistance mutations were observed in 4/86 patients (4.6%). The prevalence of HBV coinfection was high in this HIV-positive Ghanaian cohort. The two rapid assays identified HBsAg-positive patients at risk for liver disease with high specificity, albeit with only moderate sensitivity.
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Louisirirotchanakul S, Khupulsup K, Akraekthalin S, Chan KP, Saw S, Aw TC, Cho DH, Shin MG, Lim J. Comparison of the technical and clinical performance of the Elecsys HBsAg II assay with the Architect, AxSym, and Advia Centaur HBsAg screening assays. J Med Virol 2010; 82:755-62. [PMID: 20336717 DOI: 10.1002/jmv.21706] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
South East Asia has some of the highest prevalence rates of hepatitis B virus (HBV) infection (>or=8%) in the world, and the emergence of hepatitis B surface antigen (HBsAg) mutant strains is a growing problem. Assays with the highest levels of sensitivity, including mutant detection, should be used for routine HBsAg screening. In this large multicenter study, the clinical and technical performance of the fully automated Elecsys HBsAg II assay was compared with the Architect, AxSYM, and Advia Centaur HBsAg assays for HBsAg screening. Nine laboratories (three each from Thailand, Korea, and Singapore) compared the Elecsys HBsAg II assay with their routine HBsAg screening assay against a range of stored and routine clinical samples, including recombinant mutants. The Elecsys HBsAg II assay demonstrated equivalent sensitivity and specificity to the Architect HBsAg assay. However, the Elecsys HBsAg II assay recognized a native mutant sample (L94S, L97V, L98V, T123A) that the Architect HBsAg assay failed to detect. The AxSYM and Advia Centaur HBsAg assays appeared less sensitive for the detection of early HBV infection and also failed to detect some of the recombinant mutant strains. There was almost complete agreement between the Elecsys HBsAg II assay and comparator assays with respect to routine serum samples. The results of this study demonstrate that the Elecsys HBsAg II assay is a highly sensitive and specific screening assay for HBsAg and detects reliably the most important and clinically relevant HBV mutants and genotypes. It is suitable for routine HBsAg screening in Asia.
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Multicentre evaluation of the Elecsys® hepatitis B surface antigen II assay for detection of HBsAg in comparison with other commercially available assays. Med Microbiol Immunol 2009; 198:263-9. [DOI: 10.1007/s00430-009-0127-4] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/09/2009] [Indexed: 12/19/2022]
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The role of quantitative hepatitis B serology in the natural history and management of chronic hepatitis B. Hepatol Int 2009; 3:5-15. [PMID: 19763714 DOI: 10.1007/s12072-009-9149-7] [Citation(s) in RCA: 34] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/13/2009] [Revised: 07/02/2009] [Accepted: 08/06/2009] [Indexed: 12/17/2022]
Abstract
Chronic hepatitis B (CHB) remains a serious clinical problem worldwide. Advances in molecular technology have enabled the development of sensitive assays for the detection and quantification of hepatitis B virus (HBV) nucleic acid and demonstrated a positive correlation between serum HBV DNA levels and disease progression. Assessment of specific serologic and virologic factors also plays a pivotal role in the diagnosis and effective management of individuals with CHB. Recent development of quantitative assays for intrahepatic HBV replicative intermediates, as well as hepatitis B e antigen and hepatitis B surface antigen, has spurred investigation into the relationship between these factors and response to antiviral therapy and disease progression. Recent findings from preclinical and clinical investigations indicate that these factors may have promise in identifying patients likely to respond to treatment. Additional work is needed to standardize and validate these assays before they can be considered to be of true diagnostic value. Further evaluation is needed to decide which will have the greatest clinical applicability.
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Borgniet O, Parvaz P, Bouix C, Chevallier P, Trépo C, André P, Zoulim F. Clearance of serum HBsAg and anti-HBs seroconversion following antiviral therapy for chronic hepatitis B. J Med Virol 2009; 81:1336-42. [PMID: 19551826 DOI: 10.1002/jmv.21519] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
In this study, we have analyzed the evolution of serum HBsAg levels in 16 patients with chronic hepatitis B who showed an HBsAg seroconversion following antiviral therapy. The data showed that the clearance of serum HBsAg is slower than that of serum HBV DNA, which may reflect a slow kinetics of clearance of infected hepatocytes. Interestingly, HBsAg was detectable for a longer time using the Architect assay than with the Bio-Rad assay. As viremia suppression is achieved in most patients under therapy with the new generation of nucleoside analogs, these data suggest that the quantitative monitoring of serum HBsAg may represent a novel tool for the assessment of antiviral therapy efficacy.
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González R, Torres P, Castro E, Barbolla L, Candotti D, Koppelman M, Zaaijer HL, Lelie N, Allain JP, Echevarría JM. Efficacy of hepatitis B virus (HBV) DNA screening and characterization of acute and occult HBV infections among blood donors from Madrid, Spain. Transfusion 2009; 50:221-30. [PMID: 19682332 DOI: 10.1111/j.1537-2995.2009.02343.x] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
BACKGROUND Screening of blood units for hepatitis B virus (HBV) DNA identifies donations collected during the window period (WP) of the acute infection and may improve viral safety of the blood supply. It also leads to the detection of occult hepatitis B infection (OBI). STUDY DESIGN AND METHODS From January 2005 to December 2006, a total of 383,267 blood units were screened for hepatitis B surface antigen (HBsAg) and HBV DNA in two transfusion centers in Madrid, using either individual-donation nucleic acid testing (ID-NAT) or minipool (MP-NAT) of eight donations (MP8). Samples positive for HBV DNA and negative for HBsAg were confirmed by a second molecular test, the viral DNA was quantified, and a genome fragment including the region encoding the major hydrophilic region (MHR) of HBsAg was sequenced. RESULTS The overall yield of HBV DNA-positive, HBsAg-negative units was 1 in 21,282 (18 cases), higher when using ID-NAT than MP8-NAT (1:9862 vs. 1:51,011; p < 0.01). Four donations (1/95,817) were collected during the infectious pre-HBsAg WP, one during an early recovery stage, and the remaining 13 (1/29,482) were OBIs, six of whom had no detectable antibody to HBsAg. Low-level Genotype D HBV DNA was detected in all OBI cases; the frequencies of this genotype and MHR amino acid substitutions were significantly higher than reported from unselected Spanish HBsAg carriers. Donors with OBI had normal aminotransferase levels and were significantly older than donors carrying HBsAg. CONCLUSIONS Blood donors in the WP and with OBI are not uncommon in Madrid and are detected at a higher frequency with ID-NAT than MP-NAT.
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Affiliation(s)
- Rocio González
- Spanish Red Cross Blood Transfusion Center, Madrid, Spain
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Chevaliez S, Pawlotsky JM. Diagnosis and management of chronic viral hepatitis: antigens, antibodies and viral genomes. Best Pract Res Clin Gastroenterol 2008; 22:1031-48. [PMID: 19187865 DOI: 10.1016/j.bpg.2008.11.004] [Citation(s) in RCA: 49] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
Virological tools, including serological and molecular tools, are needed to diagnose chronic hepatitis B and C infections. They may also be useful to establish their prognosis, but they have found their principal application in guiding treatment decisions and assessing the virological responses to therapy. The goal of chronic hepatitis B therapy is to prevent progression of liver disease. This is achieved if HBV replication is durably abolished or significantly reduced. In HBeAg-positive patients, HBeAg clearance followed by the HBe seroconversion phase can be achieved. In HBeAg-negative patients, long-term antiviral suppression of viral replication is needed. The loss of HBsAg, eventually associated with an HBs seroconversion, is the most desirable endpoint of therapy but is rarely achieved. The efficacy endpoint of chronic hepatitis C treatment is the sustained virological response, defined by an undetectable HCV RNA in serum with a sensitive assay 24 weeks after the end of treatment. The HCV genotype and on-treatment viral kinetics can be used to tailor treatment dosages and duration.
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Affiliation(s)
- Stéphane Chevaliez
- Department of Virology & INSERM U955, French National Reference Centre for Viral Hepatitis B, C and delta, Hôpital Henri Mondor, Université Paris, Créteil, France
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Echevarría JM, Avellón A. Utilidad de la biología molecular en el diagnóstico microbiológico de las hepatitis virales. Enferm Infecc Microbiol Clin 2008; 26 Suppl 9:66-74. [DOI: 10.1016/s0213-005x(08)76543-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
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