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Tsutsumi H, Tsugawa T. Review of the past and present status of respiratory syncytial virus and rotavirus infections that commonly affect children. J Infect Chemother 2024; 30:825-831. [PMID: 38823679 DOI: 10.1016/j.jiac.2024.05.008] [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] [Received: 05/21/2024] [Accepted: 05/26/2024] [Indexed: 06/03/2024]
Abstract
Respiratory syncytial virus (RSV) and rotavirus infections are long-standing infectious diseases that affect children worldwide. RSV and rotavirus were first discovered in clinical specimens in 1955 and 1973, respectively. From their discovery to the present day, significant progress has been made in understanding these two infections. The introduction of a simple and rapid antigen diagnostic test into clinical settings in the 1990s offered new insight into the clinical characteristics and epidemiology of these infections. Regarding therapeutics, symptomatic treatments have remained the mainstay; however, prophylactic humanized anti-RSV monoclonal antibodies have been developed and advances in structural biology may allow for more effective human anti-RSV monoclonal antibodies and novel RSV vaccines to be developed soon. For rotavirus, two vaccines have been licensed and broadly applied over the past 10 years, which have been successful clinically and have changed the epidemiology of rotavirus infections in Japan.
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Affiliation(s)
| | - Takeshi Tsugawa
- Department of Pediatrics, Sapporo Medical University School of Medicine, Sapporo, Japan
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2
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Tate JE, Cortese MM, Offit PA, Parashar UD. Rotavirus Vaccines. PLOTKIN'S VACCINES 2023:1005-1024.e11. [DOI: 10.1016/b978-0-323-79058-1.00053-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/05/2025]
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Human Rotavirus Reverse Genetics Systems to Study Viral Replication and Pathogenesis. Viruses 2021; 13:v13091791. [PMID: 34578372 PMCID: PMC8473093 DOI: 10.3390/v13091791] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/14/2021] [Revised: 09/04/2021] [Accepted: 09/05/2021] [Indexed: 11/19/2022] Open
Abstract
Human rotaviruses (HuRVAs) are highly important causes of acute gastroenteritis in infants and young children worldwide. A lack of reliable and reproducible reverse genetics systems for HuRVAs has limited a proper understanding of HuRVA biology and also the rational design of live-attenuated vaccines. Since the development of the first reverse genetics system for RVAs (partially plasmid-based reverse genetics system) in 2006, there have been many efforts with the goal of generating infectious recombinant HuRVAs entirely from cloned cDNAs. However, the establishment of a HuRVA reverse genetics system was very challenging until 2019. This review article provides an overview of the historical background of the recent development of long-awaited HuRVA reverse genetics systems, beginning with the generation of recombinant human-simian reassortant RVAs with the aid of a helper virus in 2006 and the generation of recombinant animal (simian) RVAs in a helper virus-free manner in 2017, and culminating in the generation of recombinant HuRVAs entirely from plasmid cDNAs in 2019. Notably, the original HuRVA reverse genetics system has already been optimized to increase the efficiency of virus generation. Although the application of HuRVA reverse genetics systems has only just been initiated, these technologies will help to answer HuRVA research questions regarding viral replication and pathogenicity that could not be addressed before, and to develop next-generation vaccines and intestine-specific rotaviral vectors.
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Aggarwal S, Hassan E, Baldridge MT. Experimental Methods to Study the Pathogenesis of Human Enteric RNA Viruses. Viruses 2021; 13:975. [PMID: 34070283 PMCID: PMC8225081 DOI: 10.3390/v13060975] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/15/2021] [Revised: 05/18/2021] [Accepted: 05/20/2021] [Indexed: 12/16/2022] Open
Abstract
Every year, millions of children are infected with viruses that target the gastrointestinal tract, causing acute gastroenteritis and diarrheal illness. Indeed, approximately 700 million episodes of diarrhea occur in children under five annually, with RNA viruses norovirus, rotavirus, and astrovirus serving as major causative pathogens. Numerous methodological advancements in recent years, including the establishment of novel cultivation systems using enteroids as well as the development of murine and other animal models of infection, have helped provide insight into many features of viral pathogenesis. However, many aspects of enteric viral infections remain elusive, demanding further study. Here, we describe the different in vitro and in vivo tools available to explore different pathophysiological attributes of human enteric RNA viruses, highlighting their advantages and limitations depending upon the question being explored. In addition, we discuss key areas and opportunities that would benefit from further methodological progress.
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Affiliation(s)
- Somya Aggarwal
- Division of Infectious Diseases, Department of Medicine, Edison Family Center for Genome Sciences & Systems Biology, Washington University School of Medicine, St. Louis, MO 63110, USA; (S.A.); (E.H.)
| | - Ebrahim Hassan
- Division of Infectious Diseases, Department of Medicine, Edison Family Center for Genome Sciences & Systems Biology, Washington University School of Medicine, St. Louis, MO 63110, USA; (S.A.); (E.H.)
| | - Megan T. Baldridge
- Division of Infectious Diseases, Department of Medicine, Edison Family Center for Genome Sciences & Systems Biology, Washington University School of Medicine, St. Louis, MO 63110, USA; (S.A.); (E.H.)
- Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110, USA
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Komoto S, Fukuda S, Murata T, Taniguchi K. Reverse genetics system for human rotaviruses. Microbiol Immunol 2020; 64:401-406. [DOI: 10.1111/1348-0421.12795] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/26/2020] [Accepted: 04/10/2020] [Indexed: 01/02/2023]
Affiliation(s)
- Satoshi Komoto
- Department of Virology and ParasitologyFujita Health University School of Medicine Toyoake Aichi Japan
| | - Saori Fukuda
- Department of Virology and ParasitologyFujita Health University School of Medicine Toyoake Aichi Japan
| | - Takayuki Murata
- Department of Virology and ParasitologyFujita Health University School of Medicine Toyoake Aichi Japan
| | - Koki Taniguchi
- Department of Virology and ParasitologyFujita Health University School of Medicine Toyoake Aichi Japan
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Generation of Infectious Recombinant Human Rotaviruses from Just 11 Cloned cDNAs Encoding the Rotavirus Genome. J Virol 2019; 93:JVI.02207-18. [PMID: 30728265 DOI: 10.1128/jvi.02207-18] [Citation(s) in RCA: 38] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/15/2018] [Accepted: 01/28/2019] [Indexed: 12/26/2022] Open
Abstract
The generation of recombinant group A rotaviruses (RVAs) entirely from cloned cDNAs has been described only for a single animal RVA strain, simian SA11-L2. We recently developed an optimized RVA reverse genetics system based on only RVA cDNAs (11-plasmid system), in which the concentration of cDNA plasmids containing the NSP2 and NSP5 genes is 3- or 5-fold increased in relation to that of the other plasmids. Based on this approach, we generated a recombinant human RVA (HuRVA)-based monoreassortant virus containing the VP4 gene of the simian SA11-L2 virus using the 11-plasmid system. In addition to this monoreassortant virus, authentic HuRVA (strain KU) was also generated with the 11-plasmid system with some modifications. Our results demonstrate that the 11-plasmid system involving just RVA cDNAs can be used for the generation of recombinant HuRVA and recombinant HuRVA-based reassortant viruses.IMPORTANCE Human group A rotavirus (HuRVA) is a leading pathogen causing severe diarrhea in young children worldwide. In this paper, we describe the generation of recombinant HuRVA (strain KU) from only 11 cloned cDNAs encoding the HuRVA genome by reverse genetics. The growth properties of the recombinant HuRVA were similar to those of the parental RVA, providing a powerful tool for better understanding of HuRVA replication and pathogenesis. Furthermore, the ability to manipulate the genome of HuRVAs "to order" will be useful for next-generation vaccine production for this medically important virus and for the engineering of clinical vectors expressing any foreign genes.
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Wu JY, Zhou Y, Zhang GM, Mu GF, Yi S, Yin N, Xie YP, Lin XC, Li HJ, Sun MS. Isolation and characterization of a new candidate human inactivated rotavirus vaccine strain from hospitalized children in Yunnan, China: 2010-2013. World J Clin Cases 2018; 6:426-440. [PMID: 30294607 PMCID: PMC6163142 DOI: 10.12998/wjcc.v6.i11.426] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/24/2018] [Revised: 07/23/2018] [Accepted: 08/02/2018] [Indexed: 02/05/2023] Open
Abstract
AIM To determine the distribution of rotavirus VP7 gene in hospitalized children in Yunnan, China.
METHODS A total of 366 stool specimens were collected from hospitalized children in hospitals in Yunnan Province from September 2010 to December 2013. The genomic RNA electropherotypes and the G genotypes of the rotaviruses were determined. A phylogenetic analysis of the VP7 gene was performed. Rotavirus isolation was performed, and characterized by plaque, minimum essential medium, and all genes sequence analysis. Quantification of antibodies for inactivated vaccine prepared with ZTR-68 was examined by enzyme-linked immunosorbent assay and microneutralization assay.
RESULTS Group A human rotavirus was detected in 177 of 366 (48.4%) stool samples using a colloidal gold device assay. The temporal distribution of rotavirus cases showed significant correlation with the mean air temperature. Rotaviruses were isolated from 13% of the rotavirus-positive samples. The predominant genotype was G1 (43.5%), followed by G3 (21.7%), G9 (17.4%), G2 (4.3%), G4 (8.7%), and mixed (4.3%) among a total of 23 rotavirus isolates. A rotavirus strain was isolated from a rotavirus-positive stool sample of a 4-month-old child in The First People’s Hospital of Zhaotong (2010) for use as a candidate human inactivated rotavirus vaccine strain and for further research, and was designated ZTR-68. The genotype of 11 gene segments of strain ZTR-68 (RVA/Human-wt/CHN/ZTR-68/2010/G1P[8]) was characterized. The genotype constellation of strain ZTR-68 was identified as G1-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1. The VP7 and VP4 genotypes of strain ZTR-68 were similar to Wa-like strains.
CONCLUSIONS
A high prevalence of the G1, G2, and G3 genotypes was detected from 2010 to 2012. However, a dominant prevalence of the G9 genotype was identified as the cause of gastroenteritis in children in Yunnan, China, in 2013. A candidate human inactivated rotavirus vaccine strain, designated ZTR-68 was isolated, characterized, and showed immunogenicity. Our data will be useful for the future formulation and development of a vaccine in China.
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Affiliation(s)
- Jin-Yuan Wu
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Yan Zhou
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Guang-Ming Zhang
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Guo-Fa Mu
- Pediatrics Department, the First People’s Hospital of Zhaotong City, Zhaotong 657000, Yunnan Province, China
| | - Shan Yi
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Na Yin
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Yu-Ping Xie
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Xiao-Chen Lin
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Hong-Jun Li
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
| | - Mao-Sheng Sun
- Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, China
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Faecal shedding of rotavirus vaccine in Chinese children after vaccination with Lanzhou lamb rotavirus vaccine. Sci Rep 2018; 8:1001. [PMID: 29343800 PMCID: PMC5772666 DOI: 10.1038/s41598-018-19469-w] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/14/2017] [Accepted: 01/02/2018] [Indexed: 12/15/2022] Open
Abstract
Lanzhou lamb rotavirus vaccine (LLR) is an oral live attenuated vaccine first licensed in China in 2000. To date, > 60 million doses of LLR have been distributed to children. However, very little is known about faecal shedding of LLR in children. Therefore, faecal samples (n = 1,184) were collected from 114 children for 15 days post-vaccination in September–November 2011/2012. Faecal shedding and viral loads were determined by an enzyme immunoassay kit (EIA) and real-time RT-PCR. The complete genome was sequenced and the vaccine strain was isolated by culture in MA104 cells. Approximately 14.0% (16/114) of children had rotavirus-positive samples by EIA for at least 1 day post-vaccination. Viral loads in EIA-positive samples ranged from < 1.0 × 103 to 1.9 × 108 copies/g. Faecal shedding occurred as early as post-vaccination day 2 and as late as post-vaccination day 13 and peaked on post-vaccination day 5–10. One LLR strain was isolated by culture in MA104 cells. Sequence analysis showed 99% identity with LLR prototype strain. Faecal shedding of LLR in stool is common within 15 days of LLR vaccination, indicating vaccine strains can replicate in human enteric tissues.
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Kozyra I, Rzeżutka A. Farmed and companion animals as reservoirs of zoonotic rotavirus strains. POSTĘPY MIKROBIOLOGII - ADVANCEMENTS OF MICROBIOLOGY 2018; 57:156-166. [DOI: 10.21307/pm-2018.57.2.156] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/05/2025]
Abstract
Abstract
Rotavirus (RV) infections are a major epidemiological problem in humans and farm animals. So far, a number of human and animal RV strains have been identified. Based on the antigenic properties of the VP6 capsid protein, they have been classified into eight serogroups (A-H). The most important of them are viruses from group A (RVA), which are responsible for more than 90% of cases of rotaviral diarrhoea. The segmented structure of the virus genome and the presence of animals in human neighbourhood favour genetic reassortment between RV strains originating from different hosts. This could result in an emergence of zoonotic virus strains. The increasing number of human infections caused by virus strains having genotypes which have only been identified in animals indicates the need for epidemiological surveillance of infections. Additionally, the identification of epidemic virus strains in the outbreaks of disease in humans should be conducted. The identification of RVA strains circulating in humans and animals will allow the assessment of the impact of vaccination on the selection and emergence of zoonotic RVA strains.
1. Introduction. 2. General characteristics and classification of rotaviruses. 3. Group A rotavirus infection in humans. 4. Group A rotavirus infection in animals. 5. Genetic changes and reassortment as factors leading to the formation of zoonotic rotavirus strains. 6. Impact of human immunization on changes in genotype profile of circulating rotavirus strains. 7. Conclusions
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Affiliation(s)
- Iwona Kozyra
- Zakład Wirusologii Żywności i Środowiska , Państwowy Instytut Weterynaryjny – Państwowy Instytut Badawczy , Poland , Poland
| | - Artur Rzeżutka
- Zakład Wirusologii Żywności i Środowiska , Państwowy Instytut Weterynaryjny – Państwowy Instytut Badawczy , Poland , Poland
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Hashim ASM, Aboshanab KMA, El-Sayed AFM. Developing an Inactivated Rotavirus Vaccine and Evaluating the Immunogenicity Against a Commercially Available Attenuated Rotavirus Vaccine Using a Mice Animal Model. Viral Immunol 2016; 29:565-571. [PMID: 27860553 DOI: 10.1089/vim.2016.0073] [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] [Indexed: 11/12/2022] Open
Abstract
There is a high demand for public immunization against Rotavirus (RV), especially in Africa. In Africa, the attenuated RV vaccination is contraindicated in patients with immune diseases and nutrition deficiency. Therefore, the inactivated RV vaccine (IRVV) could be an alternative. In this study, we aimed to develop a pentavalent-IRVV using the most circulating RV strains in Egypt and evaluate it against the commercially available Rotarix® vaccine. Trial-IRVV was developed with 5% sucrose, 2% polysorbate-80, and adsorbed on Alum to potentiate the vaccine immune response. Then, it was injected subcutaneously into mice groups at 0-, 21-, and 35-time intervals. In parallel, Rotarix was administered twice on 0 and 28th day. The success of the pentavalent-IRVV/monovalent-Rotarix vaccine immunity rested on achieving immunoglobulin G (IgG) exceeding 1:6,400 that implies less susceptibility to RV infection (RVI). IRVV stimulating IgG >1:6,400 could be an alternative vaccination approach to reach a reasonable protective immunization level against RVI. In addition, Alum adjuvant incorporation effectively provoked a triple elevation of the immunization pattern.
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Affiliation(s)
- Ayaa S M Hashim
- 1 Research and Development Sector, Egyptian Company for Production of Vaccines, Sera and Drugs-EgyVac (VACSERA Holding Company) , Giza, Egypt
| | - Khaled M A Aboshanab
- 2 Microbiology and Immunology Department, Faculty of Pharmacy, Ain-Shams University , Abbassia, Egypt
| | - Aly F M El-Sayed
- 1 Research and Development Sector, Egyptian Company for Production of Vaccines, Sera and Drugs-EgyVac (VACSERA Holding Company) , Giza, Egypt
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12
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Do LP, Nakagomi T, Otaki H, Agbemabiese CA, Nakagomi O, Tsunemitsu H. Phylogenetic inference of the porcine Rotavirus A origin of the human G1 VP7 gene. INFECTION GENETICS AND EVOLUTION 2016; 40:205-213. [PMID: 26961591 DOI: 10.1016/j.meegid.2016.03.001] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/09/2015] [Revised: 02/25/2016] [Accepted: 03/01/2016] [Indexed: 01/15/2023]
Abstract
Rotavirus A (RVA) is an important cause of acute gastroenteritis in children worldwide. The most common VP7 genotype of human RVA is G1, but G1 is rarely detected in porcine strains. To understand the evolutionary relationships between human and porcine G1 VP7 genes, we sequenced the VP7 genes of three Japanese G1 porcine strains; the first two (PRV2, S80B) were isolated in 1980 and the third (Kyusyu-14) was isolated in 2001. Then, we performed phylogenetic and in-silico structural analyses. All three VP7 sequences clustered into lineage VI, and the mean nucleotide sequence identity between any pair of porcine G1 VP7 sequences belonging to lineage VI was 91.9%. In contrast, the mean nucleotide sequence identity between any pair of human G1 VP7 sequences belonging to lineages I-V was 95.5%. While the mean nucleotide sequence identity between any pair of porcine lineage VI strain and human lineage I-V strain was 85.4%, the VP7 genes of PRV2 and a rare porcine-like human G1P[6] strain (AU19) were 98% identical, strengthening the porcine RVA origin of AU19. The phylogenetic tree suggests that human G1 VP7 genes originated from porcine G1 VP7 genes. The time of their most recent common ancestor was estimated to be 1948, and human and porcine RVA strains evolved along independent pathways. In-silico structural analyses identified 7 amino acid residues within the known neutralisation epitopes that show differences in electric charges and shape between different porcine and human G1 strains. When compared with much divergent porcine G1 VP7 lineages, monophyletic, less divergent human G1 VP7 lineages support the hypothesis that all human G1 VP7 genes included in this study originated from a rare event of a porcine RVA transmitting to humans that was followed by successful adaptation to the human host. By contrast, AU19 represents interspecies transmission that terminated in dead-end infection.
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Affiliation(s)
- Loan Phuong Do
- Department of Hygiene and Molecular Epidemiology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan; Department of Virology, National Institute of Hygiene and Epidemiology, Hanoi, Vietnam
| | - Toyoko Nakagomi
- Department of Hygiene and Molecular Epidemiology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan; Centre for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan
| | - Hiroki Otaki
- Centre for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan
| | - Chantal Ama Agbemabiese
- Department of Hygiene and Molecular Epidemiology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan
| | - Osamu Nakagomi
- Department of Hygiene and Molecular Epidemiology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan; Centre for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
| | - Hiroshi Tsunemitsu
- Dairy Hygiene Research Division, Hokkaido Research Station, National Institute of Animal Health, Sapporo, Hokkaido, Japan
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Dormitzer PR. Rotaviruses. MANDELL, DOUGLAS, AND BENNETT'S PRINCIPLES AND PRACTICE OF INFECTIOUS DISEASES 2015:1854-1864.e4. [DOI: 10.1016/b978-1-4557-4801-3.00152-1] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/05/2025]
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Dash SK, Kumar K, Tewari A, Varshney P, Goel A, Bhatia AK. Detection of rotavirus from hospitalized diarrheic children in uttar pradesh, India. Indian J Microbiol 2013; 52:472-7. [PMID: 23997341 DOI: 10.1007/s12088-012-0279-6] [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] [Received: 07/05/2011] [Accepted: 06/04/2012] [Indexed: 11/26/2022] Open
Abstract
In the present study 220 stool samples collected from diarrheic children admitted to different hospitals and nursing homes of Uttar Pradesh and Uttarakhand were screened for rotavirus. Of 220 diarrheic samples screened 46 samples were found to be positive for rotavirus by RNA PAGE. All the isolates exhibited 4-2-3-2 migration pattern suggesting group A rotavirus. Both long and short electropherotypes were prevalent in these regions. Six different electropherotypes were detected in this study period. Male diarrheic children were found to be more susceptible to rotavirus infection (22.96 %) than that of the female ones (17.64 %). Viral RNA isolated from stool samples again subjected to VP4 gene amplification by RT-PCR using con2 and con3 primer which resulted 876 bp product suggesting group A rotavirus. Besides virus isolation was successfully done using MA104 cell line.
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Affiliation(s)
- S K Dash
- Department of Veterinary Microbiology and Immunology, College of Veterinary Sciences and Animal Husbandry, DUVASU, Mathura, 281001 Uttar Pradesh India ; Division of Virology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122 India
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Yen C, Jakob K, Esona MD, Peckham X, Rausch J, Hull JJ, Whittier S, Gentsch JR, LaRussa P. Detection of fecal shedding of rotavirus vaccine in infants following their first dose of pentavalent rotavirus vaccine. Vaccine 2011; 29:4151-5. [PMID: 21477676 DOI: 10.1016/j.vaccine.2011.03.074] [Citation(s) in RCA: 55] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/08/2010] [Revised: 03/10/2011] [Accepted: 03/22/2011] [Indexed: 01/09/2023]
Abstract
Studies on rotavirus vaccine shedding and its potential transmission within households including immunocompromised individuals are needed to better define the potential risks and benefits of vaccination. We examined fecal shedding of pentavalent rotavirus vaccine (RV5) for 9 days following the first dose of vaccine in infants between 6 and 12 weeks of age. Rotavirus antigen was detected by enzyme immunoassay (EIA), and vaccine-type rotavirus was identified by nucleotide sequencing based on genetic relatedness to the RV5 VP6 gene. Stool from 22 (21.4%) of 103 children contained rotavirus antigen-positive specimens on ≥ 1 post-vaccination days. Rotavirus antigen was detected as early as post-vaccination day 3 and as late as day 9, with peak numbers of shedding on post-vaccination days 6 through 8. Vaccine-type rotavirus was detected in all 50 antigen-positive specimens and 8 of 8 antigen-negative specimens. Nine (75%) of 12 EIA-positive and 1 EIA-negative samples tested culture-positive for vaccine-type rotavirus. Fecal shedding of rotavirus vaccine virus after the first dose of RV5 occurred over a wide range of post-vaccination days not previously studied. These findings will help better define the potential for horizontal transmission of vaccine virus among immunocompromised household contacts of vaccinated infants for future studies.
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Affiliation(s)
- Catherine Yen
- Department of Pediatrics, Columbia University, New York, NY 10032, USA
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[Diagnosis and molecular epidemiology of viral gastroenteritis in the past, present and future]. Uirusu 2010; 59:75-90. [PMID: 19927992 DOI: 10.2222/jsv.59.75] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/19/2022]
Abstract
Outline, history of research, diagnosis and molecular epidemiology of viral gastroenteritis were described. Rotavirus, adenovirus, norovirus, sapovirus, astrovirus, human parechovirus, Aichivirus, and human bocavirus are the major target viruses which cause acute gastroenteritis. The viruses were differentiated into genogroup, genotypes and subgenotypes/clusters/lineages. The changing of their genetic backgrounds was well recognized in different areas and years. Some reassortments or recombinations were observed not only between humans and humans but also between humans and animals. Viral gastroenteritis diseases were transmitted by food-borne and humans to humans contact. The environmental factors were also impacted on the infections. Recently, situation of the diseases in the natural ecosystem is becoming clearly. Diagnoses by immunological methods and gene technology are available for the known viruses. Further development of diagnosis and discovery of new viruses will be expected. Therefore, the research on molecular epidemiology is needed to be conducted continuously and then new findings will appear. We need to precede the research by using new techniques and we need to cope with the demand of society especially during acute gastroenteritis outbreak seasons.
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Arnold M, Patton JT, McDonald SM. Culturing, storage, and quantification of rotaviruses. ACTA ACUST UNITED AC 2010; Chapter 15:Unit 15C.3. [PMID: 19885940 DOI: 10.1002/9780471729259.mc15c03s15] [Citation(s) in RCA: 113] [Impact Index Per Article: 7.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/23/2022]
Abstract
Group A rotaviruses (RVs) infect the young of numerous animal species and cause acute gastroenteritis. Cultivation of animal and human RVs in cells requires proteolytic activation of the viral attachment protein using trypsin. Continuous cell lines, such as rhesus monkey kidney cells, as well as primary monkey kidney cells, are routinely used for the growth and characterization of RVs. Isolation and cultivation of human RVs from clinical fecal specimens is difficult and adaptation to growth in vitro requires multiple rounds of passage in primary cells. Following growth, RV stocks can be purified by centrifugation, if required, and quantified using plaque assay or fluorescence focus assay. This unit describes easily applicable procedures for the culturing, storage, and quantification of RVs.
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Affiliation(s)
- Michelle Arnold
- Laboratory of Infectious Diseases, NIAID/NIH, Bethesda, Maryland, USA
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Prabha S, Verghese S. Detection of porcine rotavirus from tissue and faecal specimens. Indian J Med Microbiol 2009; 27:149-52. [PMID: 19384040 DOI: 10.4103/0255-0857.49430] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/04/2022]
Abstract
Porcine small intestinal sub-mucosa is a cell-free collagen matrix that has demonstrated its ability as a scaffold material. Transplantation poses special hazards because grafted tissues and organs transmit pathogens efficiently, especially viruses. Rotavirus is thought to be confined to the intestine, causing acute diarrhoea. The purpose of this study was to evaluate the porcine intestinal tissue scaffold for Rotavirus and to study the incidence of this virus among pig herds. Only one isolate was successfully adapted to grow in cell line MA 104 from faecal samples. This isolate was further confirmed by reverse transcriptase polymerase chain reaction and sequence analysis.
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Affiliation(s)
- Suji Prabha
- Department of Microbiology, International Centre for Cardio Thoracic and Vascular Diseases, Frontier Lifeline Pvt Ltd, Dr. K.M. Cherian Heart Foundation, Chennai, India.
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20
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Gerba CP, Rose JB, Singh SN, Farrah SR. Waterborne gastroenteritis and viral hepatitis. ACTA ACUST UNITED AC 2009. [DOI: 10.1080/10643388509381732] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/21/2022]
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21
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Clark HF, Offit PA, Parashar UD, Ward RL. Rotavirus vaccines. Vaccines (Basel) 2008. [DOI: 10.1016/b978-1-4160-3611-1.50032-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/24/2022] Open
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22
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Westerman LE, Jiang B, McClure HM, Snipes-Magaldi LJ, Griffin DD, Shin G, Gentsch JR, Glass RI. Isolation and characterization of a new simian rotavirus, YK-1. Virol J 2006; 3:40. [PMID: 16737519 PMCID: PMC1524728 DOI: 10.1186/1743-422x-3-40] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/30/2005] [Accepted: 05/31/2006] [Indexed: 12/23/2022] Open
Abstract
Background To effectively analyze the requirements for protection to rotavirus infection, a reliable animal model that reasonably mimics infection and disease in humans is needed. A requirement for an effective animal model is the availability of appropriate rotavirus stocks for challenge. Results A new simian rotavirus, designated YK-1, was isolated from a 2-year-old immunodeficient pigtailed macaque with chronic diarrhea. YK-1 was distinguishable by electropherotype from the other simian rotavirus strains, SA11 and RRV. One variant of YK-1, clone 311, which was isolated after adaptation and plaque purification in cell cultures, displayed an unusual RNA electropherotype with an abnormally migrating gene 11 segment. Sequence analysis demonstrated a genetic rearrangement that involved a partial duplication of the gene 11 ORF encoding NSP5. YK-1 was identified as a Group A rotavirus belonging to subgroup 1. To further characterize the YK-1 strain, the genes encoding VP4, VP7, and NSP4 were sequenced. Analysis of VP4 and VP7 gene fragments suggests that this strain is a G3P[3] rotavirus and is closely related to the simian rotavirus strain RRV. Serotype analysis also identified YK-1 as a G3 rotavirus. The NSP4 genotype of YK-1 is C, the same genotype as RRV. Conclusion This newly isolated rotavirus, YK-1, is being used to establish a nonhuman primate model for studying the infectivity, immunity, and pathogenesis of rotavirus and for evaluating candidate rotavirus vaccines.
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Affiliation(s)
- Larry E Westerman
- Viral Gastroenteritis Team, Respiratory and Enteric Viruses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
- Yerkes National Primate Research Center, Emory University, Atlanta, Georgia, USA
| | - Baoming Jiang
- Viral Gastroenteritis Team, Respiratory and Enteric Viruses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
- Yerkes National Primate Research Center, Emory University, Atlanta, Georgia, USA
| | - Harold M McClure
- Yerkes National Primate Research Center, Emory University, Atlanta, Georgia, USA
| | - Lauren J Snipes-Magaldi
- Viral Gastroenteritis Team, Respiratory and Enteric Viruses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
| | - Dixie D Griffin
- Viral Gastroenteritis Team, Respiratory and Enteric Viruses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
| | - Gary Shin
- Department of Ecology and Evolutionary Biology, University of California at Los Angeles. Los Angeles, California, USA
| | - Jon R Gentsch
- Viral Gastroenteritis Team, Respiratory and Enteric Viruses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
| | - Roger I Glass
- Viral Gastroenteritis Team, Respiratory and Enteric Viruses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
- Yerkes National Primate Research Center, Emory University, Atlanta, Georgia, USA
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Monnier N, Higo-Moriguchi K, Sun ZYJ, Prasad BVV, Taniguchi K, Dormitzer PR. High-resolution molecular and antigen structure of the VP8* core of a sialic acid-independent human rotavirus strain. J Virol 2006; 80:1513-23. [PMID: 16415027 PMCID: PMC1346936 DOI: 10.1128/jvi.80.3.1513-1523.2006] [Citation(s) in RCA: 70] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/01/2023] Open
Abstract
The most intensively studied rotavirus strains initially attach to cells when the "heads" of their protruding spikes bind cell surface sialic acid. Rotavirus strains that cause disease in humans do not bind this ligand. The structure of the sialic acid binding head (the VP8* core) from the simian rotavirus strain RRV has been reported, and neutralization epitopes have been mapped onto its surface. We report here a 1.6-A resolution crystal structure of the equivalent domain from the sialic acid-independent rotavirus strain DS-1, which causes gastroenteritis in humans. Although the RRV and DS-1 VP8* cores differ functionally, they share the same galectin-like fold. Differences between the RRV and DS-1 VP8* cores in the region that corresponds to the RRV sialic acid binding site make it unlikely that DS-1 VP8* binds an alternative carbohydrate ligand in this location. In the crystals, a surface cleft on each DS-1 VP8* core binds N-terminal residues from a neighboring molecule. This cleft may function as a ligand binding site during rotavirus replication. We also report an escape mutant analysis, which allows the mapping of heterotypic neutralizing epitopes recognized by human monoclonal antibodies onto the surface of the VP8* core. The distribution of escape mutations on the DS-1 VP8* core indicates that neutralizing antibodies that recognize VP8* of human rotavirus strains may bind a conformation of the spike that differs from those observed to date.
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Abstract
Vaccination is the current strategy for control and prevention of severe rotavirus infections, a major cause of acute, dehydrating diarrhoea in young children worldwide. Public health interventions aimed at improving water, food and sanitation are unlikely adequately to control the disease. The development of vaccines against severe rotavirus diarrhoea is based upon homotypic or heterotypic protection provided against either a single common G serotype (monovalent vaccines) or against multiple serotypes (multivalent vaccines). Rotavirus strain surveillance has a high priority in disease control programmes worldwide. The continued identification of the most common G and P serotypes for inclusion in vaccines is an important priority. And subsequent to the introduction of a vaccine candidate, not only monitoring of circulating strains is recommended, but also surveillance of potential reassortment of animal rotavirus genes from the vaccine into human rotavirus strains is critical. Conventional methods used in the characterisation of rotavirus strains, such as enzyme immunoassay serotyping and reverse‐transcription PCR‐based genotyping often fail to identify uncommon and newly appearing strains. The application of newer molecular approaches, including sequencing and oligonucleotide microarray hybridisation, may be required to characterise such strains. The present paper presents a brief overview of the variety of standard methods available, followed by suggestions for a systematic approach for routine rotavirus strain surveillance as well as for characterisation of incompletely typed rotavirus strains. Improved detection and characterisation of incompletely typed strains will help to develop a comprehensive strain surveillance that may be required for tailoring effective rotavirus vaccines. Published in 2004 by John Wiley & Sons, Ltd.
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Affiliation(s)
- Thea K Fischer
- Centre for International Health, University of Bergen, Norway, Laboratorio National de Saúde Publica, Bissau, Guinea-Bissau and Department of Epidemiology Research, Danish Epidemiology Science Centre, Copenhagen, Denmark.
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25
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Whittier CA, Horne W, Slenning B, Loomis M, Stoskopf MK. Comparison of storage methods for reverse-transcriptase PCR amplification of rotavirus RNA from gorilla (Gorilla g. gorilla) fecal samples. J Virol Methods 2004; 116:11-7. [PMID: 14715302 DOI: 10.1016/j.jviromet.2003.10.003] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Abstract
Detection of enteric viral nucleic acids in preserved gorilla fecal specimens was investigated using reverse transcription polymerase chain reaction (rt-PCR). A commercially available viral RNA extraction kit was used to isolate nucleic acids from captive gorilla fecal samples seeded with rotavirus and stored in ethanol, formalin, a commercial RNA preservation solution, guanidine thiocyanate buffer (GT), and samples dried in tubes containing silica gel. Nucleic acids were extracted at 1, 7, 70 and 180 days and used for rt-PCR amplification of specific rotavirus RNA sequences. Successful rt-PCR amplification of the target product varied according to storage conditions, and storage time. Only samples stored in GT gave 100% positive results at 180 days. It is recommended that fecal samples be collected in GT for viral RNA analysis.
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Affiliation(s)
- Christopher A Whittier
- Environmental Medicine Consortium, College of Veterinary Medicine, North Carolina State University, 4700 Hillsborough Street, Raleigh, NC 27606, USA.
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26
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Coulson BS, Witterick PD, Tan Y, Hewish MJ, Mountford JN, Harrison LC, Honeyman MC. Growth of rotaviruses in primary pancreatic cells. J Virol 2002; 76:9537-44. [PMID: 12186936 PMCID: PMC136474 DOI: 10.1128/jvi.76.18.9537-9544.2002] [Citation(s) in RCA: 40] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023] Open
Abstract
Rotavirus infection in children at risk of developing type 1 diabetes has been temporally associated with development of pancreatic islet autoantibodies. In this study, nonobese diabetic mice were shown to be susceptible to rhesus rotavirus infection and pancreatic islets from nonobese diabetic mice, nonobese diabetes-resistant mice, fetal pigs, and macaque monkeys supported various degrees of rotavirus growth. Human rotaviruses replicated in monkey islets only. This islet susceptibility shows that rotavirus infection of the pancreas in vivo might be possible.
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Affiliation(s)
- Barbara S Coulson
- Department of Microbiology and Immunology, The University of Melbourne, Royal Parade, Victoria 3010, Australia.
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He ST, He FZ, Wu CR, Li SX, Liu WX, Yang YF, Jiang SS, He G. Treatment of rotaviral gastroenteritis with Qiwei Baizhu powder. World J Gastroenterol 2001; 7:735-40. [PMID: 11819866 PMCID: PMC4695586 DOI: 10.3748/wjg.v7.i5.735] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To observe the effects of Qiwei Baizhu Powder (QWBZP) on rotaviral gastroenteritis in children and in animal models.
METHODS: Enrolled patients were divided into two groups, and one group was treated with oral rehydration solution (ORS) and the other treated with oral liquid of QWBZP. Neonate mice were orally infected with 50 μL rotavirus suspension (4 × 108 PFU/mL) and treated with ORS or oral liquid of QWBZP, respectively.
RESULTS: Eighty-three cases of rotaviral gastroenteritis treated with QWBZP revealed a better efficacy than that treated with ORS (χ² = 10.87, P ＜ 0.05). The contents of sodium and glucose as well as number of patients with positive human rotavirus antigen in stool in QWBZP group were all less than that in ORS group. In animal models, QWBZP was found effective in treating rotavirus gastroenteritis in neonate NIH mice, as compared with control groups. In QWBZP group, the mortality of infected mice was decreased by 73.3%, the body weight of infected mice was increased, the contents of sodium and glucose as well as number of mice with positive rotavirus antigen in feces were significantly reduced, and the pathological changes such as damage of small intestinal mucosa and villi were also obviously alleviated.
CONCLUSION: QWBZP has effects on improving the absorptive function of small intestine, shortening the duration of diarrhea and rotavirus shedding from stool and alleviating the pathological changes of small intestine induced by rotavirus.
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Affiliation(s)
- S T He
- Institute of Combined Traditional Chinese and Western Medicine, Xiangya Hospital, Hunan Medical University, Changsha 410008, Hunan Province, China.
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28
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Anand T, Narasa Raju TA, Vishnu C, Venkateswar Rao LV, Sharma G. Development of Dot-ELISA for the detection of human rotavirus antigen and comparison with RNA-PAGE. Lett Appl Microbiol 2001; 32:176-80. [PMID: 11264748 DOI: 10.1046/j.1472-765x.2001.00884.x] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
AIMS Development of a simple, specific, rapid and inexpensive Dot-ELISA test for diagnosis of rotaviral antigen in stool samples. METHODS AND RESULTS Hyperimmune rabbit antisera raised against SA-11 (Simian Agent-11) strain was used as primary antibody. The secondary antibody conjugate used was the goat anti-rabbit IgG alkaline phosphatase, and BCIP/NBT solution was used as substrate. Faecal extracts were diluted 10-fold and used for the detection of rotavirus antigen. RNA-PAGE was performed to compare the specificity and sensitivity of the diagnostic tests. Dot-ELISA positive samples were further confirmed by Western blot analysis. CONCLUSIONS This Dot-ELISA test could be used as an alternative method for diagnosing rotaviral samples in the field. SIGNIFICANCE AND IMPACT OF THE STUDY The Dot-ELISA test is simple, specific, rapid and cost effective. It is suitable for identifying a large number of samples obtained from epidemiological studies and hence, reducing the death rate of rotavirus-infected patients.
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Affiliation(s)
- T Anand
- Department of Microbiology, Osmania University, Hyderabad, India
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29
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Londrigan SL, Hewish MJ, Thomson MJ, Sanders GM, Mustafa H, Coulson BS. Growth of rotaviruses in continuous human and monkey cell lines that vary in their expression of integrins. J Gen Virol 2000; 81:2203-2213. [PMID: 10950978 DOI: 10.1099/0022-1317-81-9-2203] [Citation(s) in RCA: 46] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/16/2023] Open
Abstract
Rotavirus replication occurs in vivo in intestinal epithelial cells. Cell lines fully permissive to rotavirus include kidney epithelial (MA104), colonic (Caco-2) and hepatic (HepG2) types. Previously, it has been shown that cellular integrins alpha 2 beta 1, alpha 4 beta 1 and alpha X beta 2 are involved in rotavirus cell entry. As receptor usage is a major determinant of virus tropism, the levels of cell surface expression of these integrins have now been investigated by flow cytometry on cell lines of human (Caco-2, HepG2, RD, K562) and monkey (MA104, COS-7) origin in relation to cellular susceptibility to infection with monkey and human rotaviruses. Cells supporting any replication of human rotaviruses (RD, HepG2, Caco-2, COS-7 and MA104) expressed alpha 2 beta 1 and (when tested) alpha X beta 2, whereas the non-permissive K562 cells did not express alpha 2 beta 1, alpha 4 beta 1 or alpha X beta 2. Only RD cells expressed alpha 4 beta 1. Although SA11 grew to higher titres in RD, HepG2, Caco-2, COS-7 and MA104 cells, this virus still replicated at a low level in K562 cells. In all cell lines tested, SA11 replicated to higher titres than did human strains, consistent with the ability of SA11 to use sialic acids as alternative receptors. Levels of cell surface alpha 2 integrin correlated with levels of rotavirus growth. The alpha 2 integrin relative linear median fluorescence intensity on K562, RD, COS-7, MA104 and Caco-2 cells correlated linearly with the titre of SA11 produced in these cells at 20 h after infection at a multiplicity of 0.1, and the data best fitted a sigmoidal dose-response curve (r(2)=1.00, P=0.005). Thus, growth of rotaviruses in these cell lines correlates with their surface expression of alpha 2 beta 1 integrin and is consistent with their expression of alpha X beta 2 and alpha 4 beta 1 integrins.
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Affiliation(s)
- Sarah L Londrigan
- Department of Microbiology and Immunology, The University of Melbourne, Parkville 3052, Victoria, Australia1
| | - Marilyn J Hewish
- Department of Microbiology and Immunology, The University of Melbourne, Parkville 3052, Victoria, Australia1
| | - Melanie J Thomson
- Department of Microbiology and Immunology, The University of Melbourne, Parkville 3052, Victoria, Australia1
| | - Georgina M Sanders
- Department of Microbiology and Immunology, The University of Melbourne, Parkville 3052, Victoria, Australia1
| | - Huseyin Mustafa
- Department of Gastroenterology and Clinical Nutrition, The Royal Children's Hospital, Parkville 3052, Victoria, Australia2
| | - Barbara S Coulson
- Department of Gastroenterology and Clinical Nutrition, The Royal Children's Hospital, Parkville 3052, Victoria, Australia2
- Department of Microbiology and Immunology, The University of Melbourne, Parkville 3052, Victoria, Australia1
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Fujii R, Kuzuya M, Hamano M, Ogura H, Yamada M, Mori T. Neutralization assay for human group C rotaviruses using a reverse passive hemagglutination test for endpoint determination. J Clin Microbiol 2000; 38:50-4. [PMID: 10618062 PMCID: PMC86016 DOI: 10.1128/jcm.38.1.50-54.2000] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
A novel neutralization assay for human group C rotavirus (CHRV) was developed by using a reverse passive hemagglutination (RPHA) test for endpoint determination. In this assay, the neutralization (N)-RPHA test, serial twofold dilutions of sera were mixed with a solution of CHRV that yielded an RPHA test titer of 8 at 3 days after infection. The mixtures were incubated at 37 degrees C for 1 h and were inoculated onto CaCo-2 cell monolayers in a 96-well microplate. Maintenance medium containing 100 microgram of pancreatin per ml was placed in each well. The plate was sealed with sticky plastic film and was incubated at 37 degrees C for 3 days under continuous rotation. Then, the RPHA test titer of each well was determined. The neutralization titer was expressed as the reciprocal of the maximum dilution of the serum that exhibited a fourfold (75%) or greater reduction in the RPHA test titer (8 to 2 or less). Seroconversion of neutralizing antibody was demonstrated by this method in four sets of paired serum specimens from patients with diarrheal disease caused by CHRV. The seroprevalence of CHRV in the general population in Okayama Prefecture was 26.8% by immunofluorescence and 25.5% by the N-RPHA test. The N-RPHA test described here is the first system used to assay for a neutralization antibody against CHRV and is applicable in both clinical and epidemiological settings.
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Affiliation(s)
- R Fujii
- Department of Microbiology, Okayama Prefectural Institute for Environmental Science and Public Health, Okayama 701-0212, Okayama 700-8558, Japan.
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31
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Hewish MJ, Takada Y, Coulson BS. Integrins alpha2beta1 and alpha4beta1 can mediate SA11 rotavirus attachment and entry into cells. J Virol 2000; 74:228-36. [PMID: 10590110 PMCID: PMC111532 DOI: 10.1128/jvi.74.1.228-236.2000] [Citation(s) in RCA: 138] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023] Open
Abstract
Most mammalian rotaviruses contain tripeptide amino acid sequences in outer capsid proteins VP4 and VP7 which have been shown to act as ligands for integrins alpha2beta1 and alpha4beta1. Peptides containing these sequences and monoclonal antibodies directed to these integrins block rotavirus infection of cells. Here we report that SA11 rotavirus binding to and infection of K562 cells expressing alpha2beta1 or alpha4beta1 integrins via transfection is increased over virus binding to and infection of cells transfected with alpha3 integrin or parent cells. The increased binding and growth were specifically blocked by a monoclonal antibody to the transfected integrin subunit but not by irrelevant antibodies. In our experiments, integrin activation with phorbol ester did not affect virus binding to cells. However, phorbol ester treatment of K562 parent and transfected cells induced endogenous gene expression of alpha2beta1 integrin, which was detectable by flow cytometry 16 h after treatment and quantitatively correlated with the increased level of SA11 virus growth observed after this time. Virus binding to K562 cells treated with phorbol ester 24 h previously and expressing alpha2beta1 was elevated over binding to control cells and was specifically blocked by the anti-alpha2 monoclonal antibody AK7. Virus growth in alpha4-transfected K562 cells which had also been induced to express alpha2beta1 integrin with phorbol ester occurred at a level approaching that in the permissive MA104 cell line. We therefore have demonstrated that two integrins, alpha2beta1 and alpha4beta1, are capable of acting as cellular receptors for SA11 rotavirus.
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Affiliation(s)
- M J Hewish
- Department of Microbiology and Immunology, The University of Melbourne, Parkville 3052, Victoria, Australia
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32
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Steele JC. Rotavirus. Clin Lab Med 1999. [DOI: 10.1016/s0272-2712(18)30111-2] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/28/2022]
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Elimination of viruses, phages, bacteria and Cryptosporidium by a new generation Aquaguard point-of-use water treatment unit. ACTA ACUST UNITED AC 1999. [DOI: 10.1016/s0934-8859(99)80005-4] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/23/2022]
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Buesa J, Raga JV, Colomina J, de Souza CO, Muñoz C, Gil MT. Rotavirus-specific cytotoxic T lymphocytes recognize overlapping epitopes in the amino-terminal region of the VP7 glycoprotein. Virology 1999; 257:424-37. [PMID: 10329553 DOI: 10.1006/viro.1999.9646] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Rotavirus-specific cytotoxic T lymphocytes (CTL) play an important role in the resolution of rotavirus infection. The outer capsid glycoprotein, VP7, elicits a class I MHC-restricted CTL response. Vaccinia virus recombinants expressing the VP7 genes from simian rotavirus SA11 (serotype G3) and from the RF strain of bovine rotavirus (serotype G6) were used to analyze the CTL activity to this antigen in BALB/c (H-2(d)) and C57BL/6 (H-2(b)) mice neonatally infected with homologous and heterologous rotaviruses. A vaccinia virus recombinant expressing the first amino-terminal 88 amino acids of VP7 was constructed and used to search for cross-reactive CTL against this region of the protein. By using synthetic Kb, Db, and Kd motif-fitting peptides two overlapping CTL epitopes have been identified located in the first hydrophobic domain (H1) of VP7. Splenocytes obtained from rotavirus SA11-infected C57BL/6 mice induced the strongest CTL response against target cells sensitized with a peptide containing a Kb-restricted CTL epitope (amino acids 8-16). A second Kd-restricted epitope (residues 5-13) was recognized by splenocytes derived from rotavirus-infected BALB/c mice. These findings reveal the existence of CTL epitopes in the H1 signal sequence of the VP7 glycoprotein that coexist with a CTL epitope (residues 31-40) previously described within the H2 region.
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Affiliation(s)
- J Buesa
- Hospital Clinico Universitario and School of Medicine, University of Valencia, Valencia, Spain.
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35
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Abad FX, Pintó RM, Bosch A. Flow cytometry detection of infectious rotaviruses in environmental and clinical samples. Appl Environ Microbiol 1998; 64:2392-6. [PMID: 9647805 PMCID: PMC106401 DOI: 10.1128/aem.64.7.2392-2396.1998] [Citation(s) in RCA: 29] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023] Open
Abstract
A method for the detection of infectious human rotaviruses based on infection of CaCo-2 cells and detection of infected cells by indirect immunofluorescence and flow cytometry (IIF-FC) has been developed. The technique was validated by performing a seminested reverse transcription-PCR assay with sorted cell populations. The efficiency of the procedure has been compared with that of the standard method of infection of MA104 cells and ulterior detection by IIF and optical microscopy (IIF-OM) and with that of infection of MA104 cells and detection by IIF-FC. The limit of sensitivity for the detection of the cell-adapted strain Ito(r) P13, expressed as the most probable number of cytopathogenic units, was established as 200 and 2 for MA104 and CaCo-2 cells, respectively, by the IIF-FC method. The ratio of infectious virus particles to total virus particles for a wild-type rotavirus was determined to be 1/2 x 10(6) and 1/2 x 10(4) for IIF-OM with MA104 cells and IIF-FC with CaCo-2 cells, respectively. The use of IIF-FC with CaCo-2 cells was tested with fecal and water samples and proved to be more effective than the standard procedure for rotavirus detection.
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Affiliation(s)
- F X Abad
- Department of Microbiology, University of Barcelona, Spain
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36
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Abstract
One can express this subject in terms of the ‘strategy’ that a virus ‘uses’ to survive by ensuring passage from host to host. Firstly, viruses are shed into the environment by a route and in sufficient quantities to ensure access to the mucosa of another host. In this connection spread may be more successful if symptoms are produced; if, for instance, an intestinal virus produces diarrhoea or a respiratory virus causes sneezing. The virus has next to survive exposure to adverse circumstances such as drying in the air or exposure to light in order to retain sufficient viability when it reaches the mucosa: very labile viruses, such as genital herpes, are usually transmitted by close contact of mucosa to mucosa. The mucosal secretions, and in some areas the ciliary apparatus, form a barrier to all particles, including viruses. However, viruses do traverse them and reach the plasma membrane of susceptible cells in which they then initiate infection. It is possible that viruses may be trapped by phagocytic cells and be carried by them through the mucosa.
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Affiliation(s)
- R I Glass
- Gastroenteritis Section, Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA
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Sanekata T, Kuwamoto Y, Akamatsu S, Sakon N, Oseto M, Taniguchi K, Nakata S, Estes MK. Isolation of group B porcine rotavirus in cell culture. J Clin Microbiol 1996; 34:759-61. [PMID: 8904456 PMCID: PMC228888 DOI: 10.1128/jcm.34.3.759-761.1996] [Citation(s) in RCA: 25] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/02/2023] Open
Abstract
While group A and C rotaviruses have been grown in cell culture, group B rotavirus has never been cultured. In this study we successfully isolated porcine group B rotavirus in swine kidney cells. Pancreatin treatment is essential for the propagation of group B rotavirus.
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Affiliation(s)
- T Sanekata
- Department of Veterinary Microbiology, Faculty of Agriculture, Tottori University, Japan.
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39
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Effects of Qiwei Baizhu San"Equation missing" in inhibiting replication of human rotavirus in vitro) in inhibiting replication of human rotavirus in vitro. Chin J Integr Med 1996. [DOI: 10.1007/bf02934232] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/19/2022]
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40
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Sunil-Chandra NP, Mahalingam S. Isolation and subgrouping of rotaviruses from buffalo calves in Sri Lanka. Res Vet Sci 1996; 60:187-9. [PMID: 8685545 DOI: 10.1016/s0034-5288(96)90018-1] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
Twenty-eight faecal specimens from Sri Lankan buffalo calves shown to be positive for rotavirus group A antigen were subgrouped by an enzyme-linked immunosorbent assay, by using monoclonal antibodies prepared against subgroup I and II antigens. The 13 of the 28 specimens which were classified as strongly positive belonged to subgroup I. Three of five faecal specimens inoculated on to roller cultures of MA104 cells yielded group A subgroup I rotavirus. As with other group A rotaviruses isolated from human beings and young animals, the buffalo isolates required pre-treatment with trypsin and to have trypsin incorporated in the maintenance medium, and the inoculated cell cultures had to be rolled; at least six serial passages were required before distinct rotavirus cytopathic effects were produced in the MA104 cells.
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Affiliation(s)
- N P Sunil-Chandra
- Department of Veterinary Paraclinical Studies, Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, Sri Lanka
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41
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Jourdan N, Cotte Laffitte J, Forestier F, Servin AL, Quéro AM. Infection of cultured human intestinal cells by monkey RRV and human Wa rotavirus as a function of intestinal epithelial cell differentiation. RESEARCH IN VIROLOGY 1995; 146:325-31. [PMID: 8578006 DOI: 10.1016/0923-2516(96)80595-4] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
Rotaviruses display in vivo a specific tropism for enterocytes of the small intestine. We examined here the infection of cultured human intestinal epithelial Caco-2 cells by rhesus monkey rotavirus (RRV) and human Wa rotavirus. The maximal infection of these cells was obtained when trypsin was present both in the viral inoculum before adsorption to the cells and in the culture medium during the course of cell infection. Since the differentiation process of Caco-2 cells in culture closely mimics in vivo differentiation of enterocytes along the crypt-villus axis, cell infection by RRV and Wa rotavirus was examined as a function of cell differentiation. We showed that RRV and Wa rotavirus can infect equally well both undifferentiated and differentiated Caco-2 cells.
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Affiliation(s)
- N Jourdan
- CJF INSERM 94-07, Pathogénie cellulaire et moléculaire des Microorganismes entérovirulents, Faculté de Pharmacie, Châtenay-Malabry, France
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42
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Pintó RM, Diez JM, Bosch A. Use of the colonic carcinoma cell line CaCo-2 for in vivo amplification and detection of enteric viruses. J Med Virol 1994; 44:310-5. [PMID: 7852976 DOI: 10.1002/jmv.1890440317] [Citation(s) in RCA: 89] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023]
Abstract
The use of the continuous cell line CaCo-2 as an in vivo amplification system for the detection of fastidious human enteric viruses is reported. CaCo-2 cells showed an increased sensitivity to laboratory strains of group A rotavirus 3, reovirus 3, astrovirus 1, poliovirus 1, coxsackievirus A 24, enterovirus 70, and adenovirus 5, 40 and 41, when compared to a routine host cell line for each virus. Nucleic acids from wild-type infectious rotavirus, astrovirus, and adenovirus 40 in stool samples of patients with acute gastroenteritis could be amplified after infection of CaCo-2 cells with trypsin-pre-treated virus inocula. Virus diagnosis was carried out subsequently by dot-blot hybridisation with specific cDNA probes. An amplification factor between 10 and 1,000x was obtained by infection of CaCo-2 cells, thus enabling specific detection of low numbers of a wide range of enteric viruses, and the differentiation between infectious and noninfectious particles.
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Affiliation(s)
- R M Pintó
- Department of Microbiology, University of Barcelona, Spain
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43
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Abstract
Human rotaviruses, discovered nearly 20 years ago, have been proven to be major cause of paediatric diarrhoeal disease morbidity and mortality. The clinical significance of these viruses stimulated basic studies on their biology, molecular and antigenic properties and epidemiology. General features, clinical relevance, epidemiologic pattern and laboratory diagnosis of human rotavirus infections are here reviewed.
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Affiliation(s)
- G Donelli
- Laboratorio di Ultrastrutture, Istituto Superiore di Sanità, Roma, Italy
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44
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Affiliation(s)
- Y Hoshino
- Epidemiology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
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45
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Dahling DR, Wright BA, Williams FP. Detection of viruses in environmental samples: suitability of commercial rotavirus and adenovirus test kits. J Virol Methods 1993; 45:137-47. [PMID: 8113340 DOI: 10.1016/0166-0934(93)90098-c] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023]
Abstract
Commercially marketed kits are now available for rapid viral assay of clinical specimens. This study was conducted to determine the suitability of these kits for use in environmental testing. Eight rotavirus kits and one enteric adenovirus kit were screened for sensitivity using simian rotavirus SA11, human rotavirus Wa, and adenovirus 41. The most sensitive rotavirus kit and the adenovirus kit were selected for further evaluation using virus-seeded and unseeded sewage samples. The selected rotavirus kit proved capable of detecting virus at the 10(1) PFU/ml level. The enteric adenovirus kit was similarly sensitive, detecting virus at the 10(1) TCID50/ml level. Neither kit was adversely affected by the presence of sewage. Kit assay revealed 3 of 30 unseeded sewage samples to be positive for rotavirus. Adenovirus positive samples were not detected among the 30 samples. These results were confirmed using electron microscopy. It was concluded that sensitive commercial kits could provide a reasonable alternative to cell culture for the presumptive testing of environmental samples.
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Affiliation(s)
- D R Dahling
- U.S. Environmental Protection Agency, Environmental Monitoring Systems Laboratory, Cincinnati, OH 45268
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46
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Xu Z, Hardy ME, Williams JD, Woode GN, Ramig RF. Immunodominant neutralizing antigens depend on the virus strain during a primary immune response in calves to bovine rotaviruses. Vet Microbiol 1993; 35:33-43. [PMID: 8395744 DOI: 10.1016/0378-1135(93)90114-m] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
Sera obtained from gnotobiotic calves (GC antisera) infected with bovine rotavirus strain NCDV or B223 from a previous study (Woode et al., 1987), which have different G (G6 and G10 respectively) and P serotypes, were compared for their neutralization (NT) properties to a number of human and animal rotaviruses (representing G serotype 1-6, 8-10). Two distinct patterns of neutralization were identified from these GC antisera. Of all the serotypes tested, NCDV GC antisera neutralized only B641 to a relatively high titer compared with the homologous titer, implying a narrow pattern of NT response. Analysis with reassortants indicated that the response was primarily to VP4. In contrast, B223 GC antisera neutralized most of the G serotypes tested to titers within 3-7 fold of the homologous titer, demonstrating a broad pattern of NT response. In the earlier study B223 was shown to induce a heterotypic protection against bovine rotavirus B641 (G serotype 6), and the serologic data obtained from this study indicates that a B223 vaccine might provide broad protection against several different serotypes of human and animal rotaviruses.
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Affiliation(s)
- Z Xu
- Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station 77843-4467
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47
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Hughes JH. Physical and chemical methods for enhancing rapid detection of viruses and other agents. Clin Microbiol Rev 1993; 6:150-75. [PMID: 8472247 PMCID: PMC358275 DOI: 10.1128/cmr.6.2.150] [Citation(s) in RCA: 45] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/31/2023] Open
Abstract
Viral replication events can be enhanced by physical, chemical, or heat treatment of cells. The centrifugation of cells can stimulate them to proliferate, reduce their generation times, and activate gene expression. Human endothelial cells can be activated to release cyclo-oxygenase metabolites after rocking for 5 min, and mechanical stress can stimulate endothelial cells to proliferate. Centrifugation of virus-infected cultures can increase cytopathic effects (CPE), enhance the number of infected cells, increase viral yields, and reduce viral detection times and may increase viral isolation rates. The rolling of virus-infected cells also has an effect similar to that of centrifugation. The continuous rolling of virus-infected cultures at < or = 2.0 rpm can enhance enterovirus, rhinovirus, reovirus, rotavirus, paramyxovirus, herpesvirus, and vaccinia virus CPE or yields or both. For some viruses, the continuous rolling of infected cell cultures at 96 rpm (1.9 x g) is superior to rolling at 2.0 rpm for viral replication or CPE production. In addition to centrifugation and rolling, the treatment of cells with chemicals or heat can also enhance viral yields or CPE. For example, the treatment of virus-infected cells with dimethyl sulfoxide can enhance viral transformation, increase plaque numbers and plaque size, increase the number of cells producing antigens, and increase viral yields. The infectivity of fowl plague virus is increased by 80-fold when 4% dimethyl sulfoxide is added to culture medium immediately after infection. The heat shocking of virus-infected cells also has been shown to have a stimulatory effect on the replication events of cytomegalovirus, Epstein-Barr virus, and human immunodeficiency virus. The effects of motion, chemicals, or heat treatments on viral replication are not well understood. These treatments apparently activate cells to make them more permissive to viral infection and viral replication. Perhaps heat shock proteins or stress proteins are a common factor for this enhancement phenomenon. The utility of these treatments alone or in combination with other methods for enhancing viral isolation and replication in a diagnostic setting needs further investigation.
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Affiliation(s)
- J H Hughes
- Department of Medical Microbiology & Immunology, Ohio State University, Columbus 43210
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48
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Weber B, Harms F, Selb B, Doerr HW. Improvement of rotavirus isolation in the cell culture by immune peroxidase staining. J Virol Methods 1992; 38:187-94. [PMID: 1325469 DOI: 10.1016/0166-0934(92)90109-q] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/26/2022]
Abstract
Peroxidase-labeled monoclonal antibody against rotavirus group-specific antigen (inner capsid) was used for the detection of rotavirus by immunoperoxidase staining (IPS) in trypsin-free MA104 cells within 18 h post-inoculation with clinical specimens. One hundred and twenty-one fecal samples from children with acute gastroenteritis were evaluated by IPS, conventional virus isolation in cell culture and a commercially available group A-antigen ELISA (Rotazyme II, Abbott Laboratories). Fifty-eight (47.9%) stool samples were found positive by IPS. In contrast, rotavirus was isolated from only 4 (3.3%) fecal specimens by conventional cell culture (i.e. demonstration of a cytopathogenic effect). A total of 93 (76.9%) samples were positive by ELISA. IPS permits rapid detection of rotavirus infections and detects shedding of infectious virus. The method should be useful for the investigation of nosocomial spread of rotavirus infection in hospitals, contamination of environmental surfaces and desinfectants.
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Affiliation(s)
- B Weber
- Abteilung für Medizinische Virologie, Klinikum der Johann Wolfgang Goethe-Universität Frankfurt a.M., Germany
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49
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Tsunemitsu H, Saif LJ, Jiang BM, Shimizu M, Hiro M, Yamaguchi H, Ishiyama T, Hirai T. Isolation, characterization, and serial propagation of a bovine group C rotavirus in a monkey kidney cell line (MA104). J Clin Microbiol 1991; 29:2609-13. [PMID: 1663512 PMCID: PMC270383 DOI: 10.1128/jcm.29.11.2609-2613.1991] [Citation(s) in RCA: 74] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022] Open
Abstract
A virus (designated the Shintoku strain) which was morphologically indistinguishable from group A rotaviruses was detected in the feces of adult cows with diarrhea in Japan. The virus contained 11 segments of double-stranded RNA and had an electrophoretic migration pattern in polyacrylamide gels similar to that of other group C rotaviruses (4-3-2-2). Feces containing the bovine virus reacted with antiserum to porcine group C rotavirus (Cowden strain) but not group A or B rotaviruses in immunoelectron microscopy. The virus was adapted to serial propagation in roller tube cultures of a rhesus monkey kidney cell line (MA104) by using high concentrations of trypsin. Evidence for viral replication in MA104 cell cultures was demonstrated by immunoelectron microscopy and indirect immunofluorescence by using antiserum to porcine group C rotavirus and by electrophoretic analysis of extracted viral double-stranded RNA. A significant antibody response against the isolate was detected in convalescent-phase sera of cows which excreted the virus: no increased antibody response to bovine group A rotavirus was observed. To our knowledge, this is the first isolation of a group C rotavirus from cattle.
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Affiliation(s)
- H Tsunemitsu
- Hokkaido Prefectural Shintoku Animal Husbandry Experiment Station, Japan
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50
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Kitamoto N, Ramig RF, Matson DO, Estes MK. Comparative growth of different rotavirus strains in differentiated cells (MA104, HepG2, and CaCo-2). Virology 1991; 184:729-37. [PMID: 1653495 DOI: 10.1016/0042-6822(91)90443-f] [Citation(s) in RCA: 40] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022]
Abstract
The production of viral antigen after infection of MA104, HepG2 (derived from human liver), and CaCo-2 (derived from human colon) cells with various cultivatable human and animal rotavirus strains was compared using immunofluorescence tests. All rotavirus strains examined expressed antigen in CaCo-2 cells and MA104 cells, but only some virus strains, namely, SA11-Cl3 (simian), RRV (simian), CU-1 (canine), and Ty1 (turkey), produced antigen in numbers of infected HepG2 cells comparable to infections in MA104 and CaCo-2 cells. Fl-14 (equine), OSU (porcine), NCDV (bovine), and Ch2 (chicken) strains were found to infect moderate numbers of HepG2 cells. Most human rotaviruses (representing viruses in serotypes 1, 2, 3, 4, 8, and 9), a simian rotavirus variant (SA11-4F), lapine (Ala, C-11 and R-2) viruses and porcine (Gottfried) virus infections resulted either in no detectable antigen or antigen synthesis in a low percentage of HepG2 cells. Human rotavirus isolates obtained from the stool specimens of an immunocompromised child with rotavirus antigen in his liver showed two different patterns of replication in HepG2 cells. Examination of the replication of a subset of viruses in the liver and intestinal tissues of orally infected suckling mice showed the CU-1 and Ty1 strains replicated well, while the OSU and human rotavirus strains did not. These results indicate that growth restriction in HepG2 cells is not serotype-specific, and growth of a virus in HepG2 cells does not necessarily correlate with the hepatotropic potential of a virus strain. Factors that may influence these differences of virus infectivity in HepG2 cells are discussed.
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Affiliation(s)
- N Kitamoto
- Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030
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