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Chanmanee T, Ajawatanawong P, Louisirirotchanakul S, Chotiyaputta W, Chainuvati S, Wongprompitak P. Phylogenetic analysis of two new complete genomes of the hepatitis E virus (HEV) genotype 3 from Thailand. Mol Biol Rep 2020; 47:8657-8668. [PMID: 33058031 PMCID: PMC7674359 DOI: 10.1007/s11033-020-05908-3] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2019] [Accepted: 10/08/2020] [Indexed: 02/06/2023]
Abstract
Hepatitis E virus (HEV) is a causative agent of acute viral hepatitis globally. Evolutionary phylogeny classifies the HEV into eight genotypes that correlate with the viral transmission. Only four genotypes have been proven to be responsible for transmission in humans. However, there has been no report on the genomics and genotyping of HEV in Thailand during the past ten years. Here, we identified the genotype distributions of the Thai isolates of HEV and we sequenced two HEV genomes. We screened for 18 Thai isolates of HEV from Siriraj Hospital in Bangkok, from 2014–2016. The HEV genomes were sequenced from the serum and feces of a patient. The results showed that all Thai isolates of HEV were identified as genotype 3 (HEV-3). The ORF2 and genome phylogenies suggested two subgenotypes, called 3.1 and 3.2. The Thai isolates of HEV were frequently found in the subgenotype 3.1. The genome sequences of the two Thai isolates of HEV from the serum and fecal samples of the same patient showed 91% nucleotide similarity with the HEV genotype 3. Comparisons between the HEV genome and the ORF2 phylogenies illustrated that the ORF2 tree can be used to identify HEV genotypes, but it has less phylogenetic power for the HEV evolution. The two new genome sequences of HEV-3 from Thailand could contribute valuable information to the HEV genome study. (226 words)
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Affiliation(s)
- Tipsuda Chanmanee
- Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
| | - Pravech Ajawatanawong
- Division of Bioinformatics and Data Management for Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
| | - Suda Louisirirotchanakul
- Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
| | - Watcharasak Chotiyaputta
- Division of Gastroenterology, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
| | - Siwaporn Chainuvati
- Division of Gastroenterology, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
| | - Patimaporn Wongprompitak
- Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
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2
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Pelosi E, Clarke I. Hepatitis E: a complex and global disease. EMERGING HEALTH THREATS JOURNAL 2017. [DOI: 10.3402/ehtj.v1i0.7069] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
Affiliation(s)
- E Pelosi
- Department of Microbiology and Virology, Health Protection Agency, Southeast Regional Laboratory, Southampton General Hospital, Southampton, UK; and
| | - I Clarke
- Department of Molecular Microbiology, Southampton Medical School, Southampton General Hospital, Southampton, UK
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3
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Shima R, Li TC, Sendai Y, Kataoka C, Mori Y, Abe T, Takeda N, Okamoto T, Matsuura Y. Production of hepatitis E virus-like particles presenting multiple foreign epitopes by co-infection of recombinant baculoviruses. Sci Rep 2016; 6:21638. [PMID: 26905478 PMCID: PMC4764844 DOI: 10.1038/srep21638] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/21/2015] [Accepted: 01/28/2016] [Indexed: 12/14/2022] Open
Abstract
Hepatitis E virus (HEV) causes not only endemics via a fecal-oral route but also sporadic cases via zoonotic transmission or blood transfusion. HEV-like particles (HEV-LP) produced by using a baculovirus expression system are considered a candidate for mucosal vaccines for HEV infection. In this study, we attempted to produce a chimeric HEV-LP presenting various foreign epitopes on its surface. Expression of the recombinant capsid proteins carrying a myc- or FLAG-tag inserted between amino acid residues 488 and 489, which are located in the exterior loop on the protruding domain of the HEV capsid, resulted in the production of recombinant HEV-LP. Although expression of the recombinant capsid protein carrying the HA-tag inserted at the same site failed to produce any particles, co-expression with the myc-tagged capsid protein successfully yielded a chimeric HEV-LP consisting of both recombinant capsid proteins. Immunoprecipitation analyses confirmed that the chimeric particles present these foreign epitopes on the surface. Similar results were obtained for the expression of the recombinant capsid proteins carrying neutralizing epitopes of Japanese encephalitis virus. These results suggest the chimeric HEV-LP system provides a novel vaccine carrier that can accommodate multiple neutralizing epitopes on its surface.
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Affiliation(s)
- Ryoichi Shima
- Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka , Japan.,Central Research Institute for Feed and Livestock, ZEN-NOH (National Federation of Agricultural Co-operative Associations), Ibaraki, Japan
| | - Tian Cheng Li
- Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan
| | - Yutaka Sendai
- Central Research Institute for Feed and Livestock, ZEN-NOH (National Federation of Agricultural Co-operative Associations), Ibaraki, Japan
| | - Chikako Kataoka
- Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan
| | - Yoshio Mori
- Department of Virology III, National Institute of Infectious Diseases, Tokyo, Japan
| | - Takayuki Abe
- Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka , Japan
| | - Naokazu Takeda
- Thailand-Japan Research Collaboration Center on Emerging and Re-emerging Infections (RCC-ERI), Nonthaburi, Thailand
| | - Toru Okamoto
- Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka , Japan
| | - Yoshiharu Matsuura
- Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka , Japan
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Thiry D, Mauroy A, Pavio N, Purdy MA, Rose N, Thiry E, de Oliveira-Filho EF. Hepatitis E Virus and Related Viruses in Animals. Transbound Emerg Dis 2015; 64:37-52. [PMID: 25919649 PMCID: PMC7169709 DOI: 10.1111/tbed.12351] [Citation(s) in RCA: 30] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/02/2014] [Indexed: 12/28/2022]
Abstract
Hepatitis E is an acute human liver disease in healthy individuals which may eventually become chronic. It is caused by the hepatitis E virus (HEV) and can have a zoonotic origin. Nearly 57,000 people die yearly from hepatitis E-related conditions. The disease is endemic in both developing and developed countries with distinct epidemiologic profiles. In developing countries, the disease is associated with inadequate water treatment, while in developed countries, transmission is associated with animal contact and the ingestion of raw or uncooked meat, especially liver. All human HEV are grouped into at least four genotypes, while HEV or HEV-related viruses have been identified in an increasing number of domestic and wild animal species. Despite a high genetic diversity, only one single HEV serotype has been described to date for HEV genotypes 1-4. The discovery of new HEV or HEV-related viruses leads to a continuing increase in the number of genotypes. In addition, the genome organization of all these viruses is variable with overlapping open reading frames (ORF) and differences in the location of ORF3. In spite of the role of some domestic and wild animals as reservoir, the origin of HEV and HEV-related viruses in humans and animals is still unclear. This review discusses aspects of the detection, molecular virology, zoonotic transmission and origin of HEV and HEV-related viruses in the context of 'One Health' and establishes a link between the previous and the new taxonomy of this growing virus family.
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Affiliation(s)
- D Thiry
- Veterinary Virology and Animal Viral Diseases, Department of Infectious and Parasitic Diseases, FARAH, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium
| | - A Mauroy
- Veterinary Virology and Animal Viral Diseases, Department of Infectious and Parasitic Diseases, FARAH, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium
| | - N Pavio
- UMR 1161 Virology, ANSES, Animal Health Laboratory, Maisons-Alfort, France.,UMR 1161 Virology, INRA, Maisons-Alfort, France.,UMR 1161 Virology, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France
| | - M A Purdy
- National Center for HIV/Hepatitis/STD/TB Prevention, Division of Viral Hepatitis, Centers for Disease Control and Prevention, Atlanta, GA, USA
| | - N Rose
- ANSES Laboratory of Ploufragan/Plouzané, Unit of Pig Epidemiology and Welfare, Ploufragan, France
| | - E Thiry
- Veterinary Virology and Animal Viral Diseases, Department of Infectious and Parasitic Diseases, FARAH, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium
| | - E F de Oliveira-Filho
- Veterinary Virology and Animal Viral Diseases, Department of Infectious and Parasitic Diseases, FARAH, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium
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Hepatitis E: an emerging disease. INFECTION GENETICS AND EVOLUTION 2014; 22:40-59. [PMID: 24434240 DOI: 10.1016/j.meegid.2014.01.002] [Citation(s) in RCA: 89] [Impact Index Per Article: 8.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/17/2013] [Revised: 12/09/2013] [Accepted: 01/04/2014] [Indexed: 02/07/2023]
Abstract
Currently, the infection with the hepatitis E virus represents the most frequent cause for acute hepatitis and jaundice in the world. According to WHO estimations, around two billion people, representing one third of the world's population, live in endemic areas for HEV and, therefore, are at risk of infection. In developed countries, the circulation of the virus in both human and animal (swine, boar, deer) sewage has been confirmed; however, the incidence rate is low compared to that of developing countries where outbreaks of acute hepatitis transmitted via the fecal-oral route are originated, more frequently in the flooding season or after natural disasters, combined with deficient sanitary conditions. There are currently 4 known genotypes of HEV. Genotypes 1 and 2 are isolated in all human epidemic outbreaks in developing countries, while genotypes 3 and 4 are isolated not only in humans but also in animals, in both developing and industrialized countries. These data support genotypes 3 and 4 having zoonotic nature. The diagnosis of this disease is based in the detection of anti-HEV IgG and IgM in blood serum using enzyme-linked immunosorbent methods. However, the method that best confirms the diagnosis is the RT-PCR, which detects HEV RNA in blood serum and also provides the genotype. The clinical course is generally that of an acute hepatitis which in some cases may require hospitalization and that, in transplant patients or HIV infected individuals can become a chronic hepatitis. Furthermore, the virus constitutes an important risk for pregnant women. The hepatitis E can present a wide range of symptoms, from a subclinical case to chronic liver disease with extrahepatic manifestations. For this reason, the diagnostic is challenging if no differential diagnosis is included. There is no specific antiviral drug for hepatitis E, but satisfactory results have been observed in some patients treated with pegylated interferon alfa2a and/or ribavirin. This revision is an update of all the molecular, epidemiological, clinic and preventive knowledge on this emergent disease up to date.
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Wang H, Zhang W, Ni B, Shen H, Song Y, Wang X, Shao S, Hua X, Cui L. Recombination analysis reveals a double recombination event in hepatitis E virus. Virol J 2010; 7:129. [PMID: 20546601 PMCID: PMC2901267 DOI: 10.1186/1743-422x-7-129] [Citation(s) in RCA: 33] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/25/2010] [Accepted: 06/14/2010] [Indexed: 02/05/2023] Open
Abstract
Recombination of Hepatitis E Virus (HEV) has rarely been reported. In the present study, phylogenetic and recombination analyses were performed on 134 complete HEV genomes. Three potentially significant recombination events, including both intra-genotype and one inter-genotype, were identified by recombination detection analysis. Recombination events I and II occurred intra-genotype and inter-genotype, respectively, among three isolates, including the lineage represented by CHN-XJ-SW13 (GU119961, swine isolate), E067-SIJ05C (AB369690, human isolate), and JJT-Kan (AB091394, human isolate), and lead to the recombinant swine isolate swCH31 (DQ450072). Recombination event III occurred between the lineage represented by the NA1 (M73218) and K52-87 (L25595), which resulted in the recombinant Xingjiang-1 (D11092). Our analyses proved that that recombination could occur between human and swine HEV strains, double recombination events existed in HEV, and recombination event could happen within ORF2 region of HEV. These results will provide valuable hints for future research on HEV diversity.
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Affiliation(s)
- Hua Wang
- School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, China
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7
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Pelosi E, Clarke I. Hepatitis E: a complex and global disease. EMERGING HEALTH THREATS JOURNAL 2008; 1:e8. [PMID: 22460217 PMCID: PMC3167588 DOI: 10.3134/ehtj.08.008] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 07/12/2007] [Revised: 03/12/2008] [Accepted: 04/10/2008] [Indexed: 12/13/2022]
Abstract
Thirty years after its discovery, the hepatitis E virus (HEV) continues to represent a major public health problem in developing countries. In developed countries, it has emerged as a significant cause of non-travel-associated acute hepatitis. HEV infects a wide range of mammalian species and a key reservoir worldwide appears to be swine. Genomic sequence similarity between some human HEV genotypes and swine HEV strains has been identified and we know that humans can acquire HEV infection from animals. Although for the most part the clinical course of HEV infection is asymptomatic or mild, significant risk of serious disease exists in pregnant women and those with chronic liver disease. In addition, there are data on the threat of chronic infections in immunocompromised patients. Beyond management of exposure by public health measures, recent data support that active immunisation can prevent hepatitis E, highlighting the need for vaccination programmes. Here we review the current knowledge on HEV, its epidemiology, and the management and prevention of human disease.
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Affiliation(s)
- E Pelosi
- Department of Microbiology and Virology, Health Protection Agency, Southeast Regional Laboratory, Southampton General Hospital, Southampton, UK
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Abstract
Hepatitis E virus (HEV) is the aetiological agent of non-HAV enterically transmitted hepatitis. It is the major cause of sporadic as well as epidemic hepatitis, which is no longer confined to Asia and developing countries but has also become a concern of the developed nations. In the Indian subcontinent, it accounts for 30-60% of sporadic hepatitis. It is generally accepted that hepatitis E is mostly self-limited and never progresses to chronicity. It has a higher mortality in pregnant women where the disease condition is accentuated with the development of fulminant liver disease. Currently, no antiviral drug or vaccine is licensed for HEV, although a vaccine candidate is in clinical trials. HEV genome is 7.2kb in size with three open reading frames (ORFs) and 5' and 3' cis acting elements, which have important roles to play in HEV replication and transcription. ORF1 codes for methyl transferase, protease, helicase and replicase; ORF2 codes for the capsid protein and ORF3 for a protein of undefined function. HEV has recently been classified in the genus Hepevirus of the family Hepeviridae. There are four major recognised genotypes with a single known serotype. The absence of a reliable in vitro propagation system is an obstacle to deciphering HEV biology. The genome of HEV has been cloned, sequenced and the infectious nature of these replicons has been established. However, questions related to replication, transcription, virus-host interactions and pathogenesis remain to be answered. This comprehensive review summarises the progress made so far in HEV research, and addresses some of the unanswered questions.
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Affiliation(s)
- Subrat Kumar Panda
- Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.
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9
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Okamoto H. Genetic variability and evolution of hepatitis E virus. Virus Res 2007; 127:216-28. [PMID: 17363102 DOI: 10.1016/j.virusres.2007.02.002] [Citation(s) in RCA: 230] [Impact Index Per Article: 12.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/15/2006] [Revised: 01/13/2007] [Accepted: 02/03/2007] [Indexed: 01/14/2023]
Abstract
Hepatitis E virus (HEV) is the sole member of the genus Hepevirus in the family Hepeviridae. HEV is transmitted primarily by the fecal-oral route, and water-borne epidemics are characteristic of hepatitis E in many developing countries in Asia, Africa and Latin America where sanitation conditions are suboptimal. Accumulating lines of evidence indicate that HEV-associated hepatitis also occurs domestically among individuals in industrialized countries, that there are animal reservoirs of HEV such as domestic pigs and wild boars, and that hepatitis E is a zoonosis. Based on the extensive genomic variability among HEV isolates, HEV sequences have been classified into four genotypes: genotype 1 consists of epidemic strains in developing countries in Asia and Africa; genotype 2 has been described in Mexico and several African countries; genotype 3 HEV is widely distributed and has been isolated from sporadic cases of acute hepatitis E and/or domestic pigs in many countries in the world, except for countries in Africa; and genotype 4 contains strains isolated from humans and/or domestic pigs exclusively in Asian countries. This paper reviews current knowledge on the genomic variability, geographic distribution and zoonotic aspects of HEV as well as the clinical significance of genotype and evolution of HEV.
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Affiliation(s)
- Hiroaki Okamoto
- Division of Virology, Department of Infection and Immunity, Jichi Medical University, School of Medicine, Tochigi-Ken 329-0498, Japan.
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Wei S, Xu Y, Wang M, To SST. Phylogenetic analysis of hepatitis E virus isolates in southern China (1994-1998). J Clin Virol 2006; 36:103-10. [PMID: 16621689 DOI: 10.1016/j.jcv.2006.03.001] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/21/2005] [Revised: 03/02/2006] [Accepted: 03/03/2006] [Indexed: 11/29/2022]
Abstract
BACKGROUND We have previously reported the identification of divergent hepatitis E virus (HEV) isolated (G9, G20 and 93G) in Guangzhou, a city in southern China. They are now recognised as a new HEV subgenotype in the world. However, the relatedness and significance of these novel isolates in sporadic HEV infection in southern China is still unclear. OBJECTIVES To perform phylogenetic analysis of nucleotide sequences from 41 HEV isolates in southern China from 1994 to 1998. STUDY DESIGN The partial nucleotide sequence of the HEV isolates were determined and compared with reported sequences in the GenBank. Their relatedness was analysed using computer software. RESULTS The majority of the HEV isolates, 39 out of 41, were found to belong to the Burmese-like isolates (genotype 1). The other two belonged to the Guangzhou-like isolates. The latter were only found in the samples collected in 1994. They, together with the G9 isolate, form a unique tree located between genotype 1 and genotype 4 (divergent HEV strains from northern China and Taiwan) on the phylogenetic tree. CONCLUSION Our results suggest that the Burmese-like isolates are the main causative agents of sporadic HEV infection in southern China. The Guangzhou-like isolates, which appeared transiently in 1994, did not seem to adapt to the environment and have caused no sporadic infection since.
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Affiliation(s)
- Shaojing Wei
- The Municipal Infectious Disease Hospital of Guangzhou, 627 Dong Fong East Road, Guangzhou 510060, People's Republic of China
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Lu L, Li C, Hagedorn CH. Phylogenetic analysis of global hepatitis E virus sequences: genetic diversity, subtypes and zoonosis. Rev Med Virol 2006; 16:5-36. [PMID: 16175650 DOI: 10.1002/rmv.482] [Citation(s) in RCA: 580] [Impact Index Per Article: 30.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
Nucleotide sequences from a total of 421 HEV isolates were retrieved from Genbank and analysed. Phylogenetically, HEV was classified into four major genotypes. Genotype 1 was more conserved and classified into five subtypes. The number of genotype 2 sequences was limited but can be classified into two subtypes. Genotypes 3 and 4 were extremely diverse and can be subdivided into ten and seven subtypes. Geographically, genotype 1 was isolated from tropical and several subtropical countries in Asia and Africa, and genotype 2 was from Mexico, Nigeria, and Chad; whereas genotype 3 was identified almost worldwide including Asia, Europe, Oceania, North and South America. In contrast, genotype 4 was found exclusively in Asia. It is speculated that genotype 3 originated in the western hemisphere and was imported to several Asian countries such as Japan, Korea and Taiwan, while genotype 4 has been indigenous and likely restricted to Asia. Genotypes 3 and 4 were not only identified in swine but also in wild animals such as boar and a deer. Furthermore, in most areas where genotypes 3 and 4 were characterised, sequences from both humans and animals were highly conserved, indicating they originated from the same infectious sources. Based upon nucleotide differences from five phylogenies, it is proposed that five, two, ten and seven subtypes for HEV genotypes 1, 2, 3 and 4 be designated alphabetised subtypes. Accordingly, a total of 24 subtypes (1a, 1b, 1c, 1d, 1e, 2a, 2b, 3a, 3b, 3c, 3d, 3e, 3f, 3g, 3h, 3i, 3j, 4a, 4b, 4c, 4d, 4e, 4f and 4g) were given.
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Affiliation(s)
- Ling Lu
- Division of Gastroenterology/Hepatology, Department of Medicine, Kansas University Medical Center, Kansas City, Kansas 66160, USA.
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Zhai L, Dai X, Meng J. Hepatitis E virus genotyping based on full-length genome and partial genomic regions. Virus Res 2006; 120:57-69. [PMID: 16472882 DOI: 10.1016/j.virusres.2006.01.013] [Citation(s) in RCA: 89] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/16/2005] [Revised: 12/23/2005] [Accepted: 01/10/2006] [Indexed: 01/28/2023]
Abstract
Some genomic regions for hepatitis E virus (HEV) genotyping have been reported to correlate well with the results from the phylogenetic analyses on the basis of the complete genome. However, few studies have systemically investigated the genomic regions for HEV genotyping using a combined phylogenetic and statistical approach. A consensus region for HEV genotyping has not been determined. In this study the nucleotide identities and genetic distances of 24 partial genomic regions and the complete genome sequences of 37 HEV strains were compared statistically. It was demonstrated with both one-way ANOVA and two-way ANOVA that only one genomic region in RNA-dependent RNA polymerase domain (4254-4560nt) for which there were no significant differences when compared with the full-length genome (P>0.05). The same four genotypes were identified by phylogenetic analysis based on this statistically predicted region identified as for the complete genome. RT-PCR amplification of HEV strains from all four genotypes confirmed conservation of the flanking primer sites of this region. Serum samples from 20 patients with a clinical diagnosis of hepatitis E were further analyzed by PCR using the same primers, 13 were positive and could be classified into genotype 4. These data strongly suggested that this newly identified region could be used for future HEV genotyping.
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Affiliation(s)
- Lijie Zhai
- Department of Microbiology and Immunology, School of Medicine, Southeast University, 87# Dingjiaqiao Rd., Nanjing, Jiangsu 210009, China.
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Tyagi S, Surjit M, Lal SK. The 41-amino-acid C-terminal region of the hepatitis E virus ORF3 protein interacts with bikunin, a kunitz-type serine protease inhibitor. J Virol 2005; 79:12081-7. [PMID: 16140784 PMCID: PMC1212588 DOI: 10.1128/jvi.79.18.12081-12087.2005] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
Hepatitis E virus (HEV), a human plus-stranded RNA virus, contains three open reading frames (ORF). Of these, ORF1 encodes the viral nonstructural polyprotein, ORF2 encodes the major capsid protein, and ORF3 codes for a phosphoprotein of undefined function. Recently, using the yeast two-hybrid system to screen a human cDNA liver library, we have isolated and characterized AMBP (alpha1-microglobulin/bikunin precursor), which specifically interacts with the ORF3 protein of HEV. The ORF3 protein expedites the processing and secretion of alpha1-microglobulin. When checked individually for interaction, the second processed protein from AMBP, bikunin, strongly interacted with the full-length ORF3 protein. This protein-protein interaction has been validated by immunoprecipitation in both COS-1 and Huh7 cells and by His6 pull-down assays. In dual-labeling immunofluorescent staining, followed by fluorescence microscopy of transfected human liver cells, ORF3 colocalized with endogenously expressed bikunin. Finally, a 41-amino-acid C-terminal region of ORF3 has been found to be responsible for interacting with bikunin. The importance of this virus-host protein-protein interaction, with reference to the viral life cycle, has been discussed.
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Affiliation(s)
- Shweta Tyagi
- Virology Group, International Centre for Genetic Engineering & Biotechnology, P. O. Box 10504, Aruna Asaf Ali Road, New Delhi 10067, India
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Lu L, Drobeniuc J, Kobylnikov N, Usmanov RK, Robertson BH, Favorov MO, Margolis HS. Complete sequence of a Kyrgyzstan swine hepatitis E virus (HEV) isolated from a piglet thought to be experimentally infected with human HEV. J Med Virol 2005; 74:556-62. [PMID: 15484284 DOI: 10.1002/jmv.20214] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
Hepatitis E virus (HEV) was identified by RT-PCR amplification with degenerate ORF2 primers in the stool of a piglet experimentally inoculated with a stool suspension from a patient with acute hepatitis during an outbreak of non-A, non-B hepatitis in Kyrgyzstan. Further characterization by sequencing of the complete genome and phylogenetic analysis showed that the piglet isolate was most closely related to HEV genotype 3. Because the original human stool specimen used to inoculate the piglet was no longer available, stool samples from three patients obtained during the same outbreak were sequenced and found to be HEV genotype 1. These findings suggest that the HEV isolated from the swine stool was probably an HEV enzootic in Kyrgyzstan and not the virus inoculated from the human stool.
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Affiliation(s)
- Ling Lu
- Laboratory Branch, Division of Viral Hepatitis, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA
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15
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Chen GB, Meng JH. Identification of 5’ capped structure and 3’ terminal sequence of hepatitis E virus isolated from Morocco. World J Gastroenterol 2004; 10:2045-9. [PMID: 15237431 PMCID: PMC4572330 DOI: 10.3748/wjg.v10.i14.2045] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
AIM: To examine 5’ and 3’ terminal sequences of hepatitis E virus (HEV) isolated from Morocco, to confirm 5’ methylated cap structure of the genome, and to investigate whether the 3’ UTR can be used to distinguish HEV genotypes instead of HEV complete genome sequence.
METHODS: RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) was employed to obtain the 5’ and 3’ terminal sequences of HEV Morocco strain. The 3’ UTR sequence of the Morocco strain was compared with that of the other 29 HEV strains using the DNAStar software.
RESULTS: The 5’ PCR product was obtained only from the RLM-RACE based on the capped RNA template. The 5’ UTR of the Morocco strain had 26 nucleotides, and the 3’ UTR had 65 nucleotides upstream to the polyA. The 5’ UTR between HEV strains had only point mutations of nucleotides. The phylogenetic tree based on the sequences of 3’ UTR was not the same as that based on the complete sequences.
CONCLUSION: The genome of HEV Morocco strain was methylated cap structure. The 3’ terminal sequence can not be used for distinguishing HEV genotype for all HEV strains in place of the whole HEV genome sequence.
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Affiliation(s)
- Guo-Bing Chen
- Department of Microbiology and Immunology, Southeast University School of Medicine, Nanjing 210009, Jiangsu Province, China
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van Cuyck H, Juge F, Roques P. Phylogenetic analysis of the first complete hepatitis E virus (HEV) genome from Africa. ACTA ACUST UNITED AC 2003; 39:133-9. [PMID: 14625096 DOI: 10.1016/s0928-8244(03)00241-4] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Abstract
Hepatitis E virus (HEV) is globally distributed, transmitted enterically and between humans and animals. Phylogenetic analysis has identified five distinct HEV genotypes. The first full-length sequence of an African strain (Chad) is presented and compared to 31 complete HEV genomes available, including the fulminant hepatitis strain from India, swine strains and a strain from Morocco. The two African strains are more closely related to genotype 1 than to any other genotypes and together they possibly form a sub-genotype or sixth genotype. The first evidence for recombination between divergent HEV strains is presented.
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Affiliation(s)
- Hélène van Cuyck
- Weapons of Mass Destruction Centre (WMD Centre), NATO, AB310, Bd. Leopold III, 1110, Brussels, Belgium.
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17
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He J, Innis BL, Shrestha MP, Clayson ET, Scott RM, Linthicum KJ, Musser GG, Gigliotti SC, Binn LN, Kuschner RA, Vaughn DW. Evidence that rodents are a reservoir of hepatitis E virus for humans in Nepal. J Clin Microbiol 2002; 40:4493-8. [PMID: 12454141 PMCID: PMC154618 DOI: 10.1128/jcm.40.12.4493-4498.2002] [Citation(s) in RCA: 44] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Hepatitis E virus (HEV) is an important cause of enterically transmitted hepatitis in developing countries. Sporadic autochthonous cases of hepatitis E have been reported recently in the United States and other industrialized countries. The source of HEV infection in these cases is unknown; zoonotic transmission has been suggested. Antibodies to HEV have been detected in many animals in areas where HEV is endemic and in domestic swine and rats in the United States. There is evidence supporting HEV transmission between swine and humans. Nevertheless, HEV has not been detected in wild rodents. We tested murid rodents and house shrews trapped in Nepal's Kathmandu Valley, where hepatitis E is hyperendemic, for HEV infection. The most commonly trapped species was Rattus rattus brunneusculus. Serum samples from 675 animals were tested for immunoglobulin G against HEV by enzyme-linked immunosorbent assay; 78 (12%) were positive, indicating acute or past infection. Antibody prevalence was higher among R. rattus brunneusculus and Bandicota bengalensis than in Suncus murinus. Forty-four specimens from 78 antibody-positive animals had sufficient residual volume for detection of HEV RNA (viremia) by reverse transcription-PCR. PCR amplification detected four animals (9%; three were R. rattus brunneusculus and one was B. bengalensis) with viremia. Phylogenetic analysis of the four genome sequences (405 bp in the capsid gene) recovered showed that they were identical, most closely related to two human isolates from Nepal (95 and 96% nucleotide homology, respectively), and distinct from HEV sequences isolated elsewhere. These data prove that certain peridomestic rodents acquire HEV in the wild and suggest that cross-species transmission occurs, with rodents serving as a virus reservoir for humans.
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Affiliation(s)
- Junkun He
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Bruce L. Innis
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Mrigendra P. Shrestha
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Edward T. Clayson
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Robert M. Scott
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Kenneth J. Linthicum
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Guy G. Musser
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Scott C. Gigliotti
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Leonard N. Binn
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - Robert A. Kuschner
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
| | - David W. Vaughn
- Walter Reed Army Institute of Research, Silver Spring, Maryland, Walter Reed Army Institute of Research/Armed Forces Research Institute of Medical Sciences Research Unit—Nepal, Kathmandu, Nepal, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, American Museum of Natural History, New York, New York
- Corresponding author. Present address: Military Infectious Diseases Research Program, U.S. Army Medical Research and Materiel Command, 504 Scott St., Fort Detrick, MD 21702-5012. Phone: (301) 619-7882. Fax: (301) 619-2416. E-mail:
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18
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Okamoto H, Takahashi M, Nishizawa T, Fukai K, Muramatsu U, Yoshikawa A. Analysis of the complete genome of indigenous swine hepatitis E virus isolated in Japan. Biochem Biophys Res Commun 2001; 289:929-36. [PMID: 11741279 DOI: 10.1006/bbrc.2001.6088] [Citation(s) in RCA: 158] [Impact Index Per Article: 6.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
Using reverse transcription-polymerase chain reaction with primers derived from well-conserved genomic areas among all four hepatitis E virus (HEV) genotypes (I-IV), the HEV sequence was identified in serum samples obtained from 3 (3%) out of 95 60- to 90-day-old pigs in Japan and characterized molecularly. In the partial sequence of open reading frame (ORF) 2 of 421 nucleotides, the three swine isolates (swJ570, swJ681, and swJ791) showed the highest similarity of 83-87% to genotype III HEV representing human and swine strains (US1, US2, and swUS1) in the United States. The full-length nucleotide sequence of swJ570 consisted of 7225 nucleotides excluding the poly(A) tail and contained ORF 1 encoding 1703 amino acids (aa), ORF2 encoding 660 aa, and ORF3 encoding 122 aa. The swJ570 strain was most closely related to a Japanese strain (JRA1), which had been obtained from a hepatitis patient who had not traveled outside Japan. The overall nucleotide sequence identity between them was 89% and the deduced amino acid sequence identities of ORF1, ORF2, and ORF3 were 96, 99, and 98%, respectively. These results indicate that a certain proportion of pigs in Japan are HEV-viremic and may act as reservoirs of HEV infection, and that the presence of an indigenous strain(s) of HEV should be taken into consideration for the diagnosis of acute hepatitis in Japan.
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Affiliation(s)
- H Okamoto
- Immunology Division and Division of Molecular Virology, Jichi Medical School, Tochigi-Ken, 329-0498, Japan.
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19
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Zhang M, Purcell RH, Emerson SU. Identification of the 5' terminal sequence of the SAR-55 and MEX-14 strains of hepatitis E virus and confirmation that the genome is capped. J Med Virol 2001; 65:293-5. [PMID: 11536235 DOI: 10.1002/jmv.2032] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/07/2022]
Abstract
Hepatitis E virus (HEV) is a nonenveloped virus with a genome of single-stranded, positive-sense RNA. The 5' terminal sequence of two HEV strains (SAR-55 and MEX-14) was determined by a 5' RNA ligase-mediated rapid amplification of cDNA ends (RACE) method designed to select capped RNAs. The 5' noncoding region of the SAR-55 and MEX-14 strains were amplified, confirming that the genomic RNA of HEV is capped. The 5' noncoding region of the SAR-55 strain had 25 nucleotides, which is two less than reported for the Burmese strain, and that of the MEX-14 strain had 24 nucleotides, which is 21 more than reported previously [Huang et al., 1992].
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Affiliation(s)
- M Zhang
- Hepatitis Viruses and Molecular Hepatitis Sections, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA
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20
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Haqshenas G, Shivaprasad HL, Woolcock PR, Read DH, Meng XJ. Genetic identification and characterization of a novel virus related to human hepatitis E virus from chickens with hepatitis-splenomegaly syndrome in the United States. J Gen Virol 2001; 82:2449-2462. [PMID: 11562538 DOI: 10.1099/0022-1317-82-10-2449] [Citation(s) in RCA: 252] [Impact Index Per Article: 10.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023] Open
Abstract
Hepatitis-splenomegaly (HS) syndrome is an emerging disease in chickens in North America; the cause of this disease is unknown. In this study, the genetic identification and characterization of a novel virus related to human hepatitis E virus (HEV) isolated from bile samples of chickens with HS syndrome is reported. Based upon the similar genomic organization and significant sequence identity of this virus with HEV, the virus has been tentatively named avian HEV in order to distinguish it from human and swine HEV. Electron microscopy revealed that avian HEV is a non-enveloped virus particle of 30-35 nm in diameter. The sequence of the 3' half of the viral genome ( approximately 4 kb) was determined. Sequence analyses revealed that this genomic region contains the complete 3' non-coding region, the complete genes from open reading frames (ORFs) 2 and 3, the complete RNA-dependent RNA polymerase (RdRp) gene and a partial helicase gene from ORF 1. The helicase gene is the most conserved gene between avian HEV and other HEV strains, displaying 58-61% aa and 57-60% nt sequence identities. The RdRp gene of avian HEV shares 47-50% aa and 52-53% nt sequence identities and the putative capsid gene (ORF 2) of avian HEV shares 48-49% aa and 48-51% nt sequence identities with the corresponding regions of other known HEV strains. Phylogenetic analysis indicates that avian HEV is genetically related to, but distinct from, other known HEV strains. This discovery has important implications for HEV animal models, nomenclature and natural history.
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Affiliation(s)
- G Haqshenas
- Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 1410 Price's Fork Road, Blacksburg, VA 24061-0342, USA1
| | - H L Shivaprasad
- California Animal Health and Food Safety Laboratory System, School of Veterinary Medicine, University of California-Davis, 2789 South Orange Avenue, Fresno, CA 93725, USA2
| | - P R Woolcock
- California Animal Health and Food Safety Laboratory System, School of Veterinary Medicine, University of California-Davis, 2789 South Orange Avenue, Fresno, CA 93725, USA2
| | - D H Read
- California Veterinary Diagnostic Laboratory System, School of Veterinary Medicine, University of California-Davis, San Bernardino, CA 92408, USA3
| | - X J Meng
- Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 1410 Price's Fork Road, Blacksburg, VA 24061-0342, USA1
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21
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Im SW, Zhang JZ, Zhuang H, Che XY, Zhu WF, Xu GM, Li K, Xia NS, Ng MH. A bacterially expressed peptide prevents experimental infection of primates by the hepatitis E virus. Vaccine 2001; 19:3726-32. [PMID: 11395207 DOI: 10.1016/s0264-410x(01)00100-1] [Citation(s) in RCA: 53] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
A 23 kDa peptide of the major structural protein of the hepatitis E virus (HEV) expressed in E. coli was found to naturally interact with one another to form homodimers and the peptide was recognized strongly in its dimeric form by HEV reactive human sera. To determine if the peptide may confer protection against HEV infection, three monkeys were immunized with the purified peptide and three were given placebo. Both groups of animals were challenged with 10(5) genome equivalent dose of the homologous strain of HEV. All control animals excreted the virus for 10-12 days beginning 5 days after the infection. The viral genome was also present in the peripheral blood monocyte (PBMC) samples from two animals, but it was not detected in the plasma samples from any of the animals. The infection in two control animals was accompanied by HEV seroconversion. Immunization was found to abrogate HEV stool excretion in two animals and reduced the viral excretion to one day in the third. None of the immunized animals showed detectable HEV in plasma or PBMC samples nor did the animals showed evidence of HEV seroconversion. These results suggested that immunization with the bacterially expressed peptide may prevent experimental infection of primates with the homologous strain of HEV.
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Affiliation(s)
- S W Im
- Department of Microbiology, Faculty of Medicine, The University of Hong Kong, Queen Mary Hospital Compound SAR, Hong Kong, People's Republic of China
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22
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Pina S, Buti M, Cotrina M, Piella J, Girones R. HEV identified in serum from humans with acute hepatitis and in sewage of animal origin in Spain. J Hepatol 2000; 33:826-33. [PMID: 11097493 DOI: 10.1016/s0168-8278(00)80316-5] [Citation(s) in RCA: 180] [Impact Index Per Article: 7.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
BACKGROUND/AIMS Hepatitis E virus (HEV) is an enterically transmitted pathogen that appears sporadically in non-endemic countries. We studied HEV as a causal agent of acute hepatitis cases in the Spanish population, and the role of pigs as an animal reservoir. METHODS The presence of HEV-RNA was analysed by nested polymerase chain reaction in 37 serum samples from patients with acute viral hepatitis, 48 porcine serum samples, 6 pig faecal samples and 12 slaughter-house sewage samples. Presence of antibodies was also tested in porcine sera. RESULTS HEV-RNA was found in 3 human serum samples from patients presenting IgG anti-HEV antibodies. Nucleotide sequence analysis identified 2 strains with 93.4% identity, phylogenetically most closely related to the Greece1 isolate, and more closely related to North American and other European strains than to those from endemic regions. HEV-RNA was also detected in slaughterhouse sewage mainly from pigs, presenting 92-94% nucleotide similarity compared to the strains detected in the human sera. Twenty-five per cent of the pigs tested presented IgG anti-HEV antibodies. CONCLUSIONS These data suggest that the HEV could be more widespread than previously thought, and present new evidence of the close relationship between HEV strains detected in pigs and those from acute hepatitis patients.
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Affiliation(s)
- S Pina
- Department of Microbiology, University of Barcelona, Spain
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23
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Wang Y, Zhang H, Ling R, Li H, Harrison TJ. The complete sequence of hepatitis E virus genotype 4 reveals an alternative strategy for translation of open reading frames 2 and 3. J Gen Virol 2000; 81:1675-86. [PMID: 10859372 DOI: 10.1099/0022-1317-81-7-1675] [Citation(s) in RCA: 142] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022] Open
Abstract
Isolates of hepatitis E virus (HEV) have recently been described from China that are distinct from Burmese, Mexican and US viruses and constitute a novel genotype (genotype 4). Here, the complete genomic sequence of a representative isolate of genotype 4 HEV, amplified directly from the stool of an acutely infected patient, is presented. Analysis of the entire sequence confirms our previous conclusion, based upon partial sequence data, that these Chinese isolates belong to a novel genotype. Typical of genetic variation in HEV, most nucleotide substitutions occur in the third base of the codon and do not affect the amino acid sequence. The genotype 4 virus is unusual in that a single nucleotide insertion in the ORF 3 region changes the initiation of ORF 3, and perhaps also ORF 2. The consequences of these changes are discussed.
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Affiliation(s)
- Y Wang
- Department of Medicine, Royal Free and University College Medical School, University College London, Royal Free Campus, Rowland Hill Street, London NW3 2PF, UK
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24
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He J, Binn LN, Tsarev SA, Hayes CG, Frean JA, Isaacson M, Innis BL. Molecular characterization of a hepatitis E virus isolate from Namibia. J Biomed Sci 2000; 7:334-8. [PMID: 10895057 DOI: 10.1007/bf02253253] [Citation(s) in RCA: 22] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/25/2022] Open
Abstract
Hepatitis E virus (HEV) causes sporadic and epidemic acute viral hepatitis in many developing countries. In Africa, hepatitis E has been documented by virus detection (reverse transcriptase polymerase chain reaction, RT-PCR) in Egypt, Chad, Algeria, Morocco and Tunisia. Cases of presumptive hepatitis E also have been documented by detection of antibody to HEV in the Sudan, Kenya, Ethiopia, Somalia, Djibouti and South Africa. Recently, we reported the recovery of 9 isolates of HEV from feces collected during an outbreak of jaundice in Namibia. These specimens were stored frozen for many years at the South African Institute for Medical Research awaiting new methods to determine the etiology of jaundice. HEV genomic sequences were detected by antigen-capture RT-PCR with primers that amplified 2 independent regions of the HEV genome (ORF-2 and ORF-3). To further characterize the HEV 83-Namibia isolates, we determined the nucleotide (nt) sequence of the 3' end of the capsid gene (296 of 1, 980 nt in ORF-2) and ORF-3 for 1 isolate. The capsid gene sequence shared 86% identity with the prototype Burma strain and up to 96% identity with other African strains at the (nt) level, and 99% identity with Burma or other Africa strains at the amino acid level. A 188 (nt) fragment amplified from ORF-3 was also highly homologous to other HEV but was too short for meaningful comparison. Phylogenetic analysis indicated that HEV 83-Namibia is closely related to other African isolates, and differs from Burmese, Mexican and Chinese HEV. These data link the HEV causing the 1983 Namibia outbreak to more recent HEV transmission in northern and sub-Saharan Africa, suggesting this subgenotype of HEV is firmly established throughout the continent.
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Affiliation(s)
- J He
- Department of Virus Diseases, Walter Reed Army Institute of Research, Walter Reed Army Medical Center, Silver Spring, MD 20910, USA.
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25
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Schofield DJ, Glamann J, Emerson SU, Purcell RH. Identification by phage display and characterization of two neutralizing chimpanzee monoclonal antibodies to the hepatitis E virus capsid protein. J Virol 2000; 74:5548-55. [PMID: 10823861 PMCID: PMC112041 DOI: 10.1128/jvi.74.12.5548-5555.2000] [Citation(s) in RCA: 129] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/29/2023] Open
Abstract
Two monoclonal antibodies (MAbs) against the ORF2 protein of the SAR-55 strain of hepatitis E virus (HEV) were isolated by phage display from a cDNA library of chimpanzee (Pan troglodytes) gamma1/kappa antibody genes. Both MAbs, HEV#4 and HEV#31, bound to reduced, denatured open reading frame 2 (ORF2) protein in a Western blot, suggesting that they recognize linear epitopes. The affinities (equilibrium dissociation constants, K(d)) for the SAR-55 ORF2 protein were 1.7 nM for HEV#4 and 5.4 nM for HEV#31. The two MAbs also reacted in an enzyme-linked immunosorbent assay with recombinant ORF2 protein from a heterologous HEV, the Meng strain. Each MAb blocked the subsequent binding of the other MAb to homologous ORF2 protein in indirect competition assays, suggesting that they recognize the same or overlapping epitopes. Radioimmunoprecipitation assays suggested that at least part of the linear epitope(s) recognized by the two MAbs is located between amino acids 578 and 607. MAbs were mixed with homologous HEV in vitro and then inoculated into rhesus monkeys (Macaca mulatta) to determine their neutralizing ability. Whereas all control animals developed hepatitis (elevated liver enzyme levels in serum) and seroconverted to HEV, those receiving an inoculum incubated with either HEV#4 or HEV#31 were not infected. Therefore, each MAb neutralized the SAR-55 strain of HEV in vitro.
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MESH Headings
- Amino Acid Sequence
- Animals
- Antibodies, Monoclonal/chemistry
- Antibodies, Monoclonal/genetics
- Antibodies, Monoclonal/immunology
- Antibodies, Monoclonal/metabolism
- Antibodies, Viral/chemistry
- Antibodies, Viral/genetics
- Antibodies, Viral/immunology
- Antibodies, Viral/metabolism
- Antibody Affinity
- Antibody Specificity
- Antigens, Viral/chemistry
- Antigens, Viral/genetics
- Antigens, Viral/immunology
- Binding, Competitive
- Blotting, Western
- Capsid/chemistry
- Capsid/immunology
- Cross Reactions
- Deoxyribonucleases, Type II Site-Specific/metabolism
- Epitope Mapping
- Hepatitis E/immunology
- Hepatitis E/prevention & control
- Hepatitis E/virology
- Hepatitis E virus/genetics
- Hepatitis E virus/immunology
- Hepatitis E virus/physiology
- Immunization, Passive
- Immunoglobulin Fab Fragments/chemistry
- Immunoglobulin Fab Fragments/genetics
- Immunoglobulin Fab Fragments/immunology
- Immunoglobulin Fab Fragments/metabolism
- Macaca mulatta
- Molecular Sequence Data
- Neutralization Tests
- Open Reading Frames
- Pan troglodytes/immunology
- Peptide Library
- Protein Denaturation
- Sequence Analysis
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Affiliation(s)
- D J Schofield
- Hepatitis Viruses, National Institutes of Health, Bethesda, Maryland 20852, USA.
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26
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Buisson Y, Grandadam M, Nicand E, Cheval P, van Cuyck-Gandre H, Innis B, Rehel P, Coursaget P, Teyssou R, Tsarev S. Identification of a novel hepatitis E virus in Nigeria. J Gen Virol 2000; 81:903-9. [PMID: 10725415 DOI: 10.1099/0022-1317-81-4-903] [Citation(s) in RCA: 80] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
Sporadic cases of acute hepatitis E among ten native Nigerian adults were reported in Port-Harcourt (Nigeria). Hepatitis E virus (HEV) was detected in serum and/or faecal samples of seven patients by RT-PCR of the open reading frame (ORF)-1 polymerase region and the 3'-end of ORF2. Restriction analysis widely used to distinguish genotypes I and III showed that all Nigerian strains have a pattern similar to the Mexican strain (NotI, nt 286; SmaI, nt 397; no KpnI restriction site) but displayed a BsmI restriction site at nt 213 as do most African HEV strains sequenced so far. Sequence analysis performed from internal ORF1 and ORF2 PCR products displayed strong homogeneity between the HEV isolates, determining a regional cluster. Phylogenetic analysis of nucleotide sequences revealed that these strains were more related to the Mexican prototype genotype III (87% homology in ORF1, 80% homology in ORF2) than to either the African strain genotype I (74% homology in ORF1, 77% homology in ORF2) or the USA strain genotype II (75% homology in ORF1, 77% homology in ORF2). Genetic divergence up to 15% in ORF2 with the Mexican genotype clearly defined a new subgenotype within genotype III. At the amino acid level, Nigerian strains showed more homology with genotype III (96%) than with genotype I (92%). This study clearly determined the co-existence of genotypes I and III in Africa. These Nigerian HEV strains belonging to genotype III, but sharing some properties with genotype I, could be one of the missing links between African and Latin American HEV and could help us to determine the phylogenetic evolution of HEV from the ancestral virus.
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Affiliation(s)
- Y Buisson
- Hôpital d'Instruction des Armées Val-de-Grâce, Laboratoire de biologie clinique, 74 boulevard de Port Royal, 75230 Paris cedex 05, France
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27
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Abstract
Microbial pathogens discovered as aetiological agents of human disease over the last 25 years are reviewed. Strengthening of laboratory and public health surveillance is of paramount importance for early detection and management of emerging infectious diseases.
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Affiliation(s)
- U Desselberger
- Clinical Microbiology and Public Health Laboratory, Addenbrooke's Hospital, Cambridge, UK
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28
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Zanetti AR, Schlauder GG, Romanò L, Tanzi E, Fabris P, Dawson GJ, Mushahwar IK. Identification of a novel variant of hepatitis E virus in Italy. J Med Virol 1999; 57:356-60. [PMID: 10089046 DOI: 10.1002/(sici)1096-9071(199904)57:4<356::aid-jmv5>3.0.co;2-d] [Citation(s) in RCA: 121] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
Hepatitis E infection is typically associated with areas in which hepatitis E virus (HEV) is endemic. Except for a few cases in Europe and in the United States, acute hepatitis E is usually associated with travel to endemic areas. We set out to determine the etiologic role of HEV in acute non-A-C hepatitis in Italy. The presence of HEV-RNA and antibody was determined in 218 patients diagnosed with acute viral non-A-C hepatitis. Acute hepatitis E infection was defined by the presence of HEV-RNA in sera and positivity for IgM anti-HEV and seroconversion to IgG anti-HEV. Acute hepatitis E was found in 10.1% of the patients with acute non-A-C, with 95.5% exhibiting a benign course. A more severe course was observed in a patient co-infected with HAV and HEV. Most cases were travelers to endemic areas, although 18.2% reported no travel. One patient was from a household with an infected patient. Sequence analyses of the polymerase chain reaction (PCR) product derived from a patient who never visited endemic areas, identified an isolate that is divergent significantly from all reported isolates of HEV (79.5-85.8% nucleotide identity). Evidence from this study suggests that HEV accounts for approximately 10% of acute non-A-C viral hepatitis in Italy, diagnosed generally in travelers returning from endemic areas. However, the identification of a new HEV variant in an individual who never indicated travel or contact with individuals associated with endemic areas, suggests that this virus may be native to Italy.
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Affiliation(s)
- A R Zanetti
- Institute of Virology, University of Milan, Italy.
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29
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Meng J, Cong ME, Dai X, Pillot J, Purdy MA, Fields HA, Khudyakov YE. Primary structure of open reading frame 2 and 3 of the hepatitis E virus isolated from Morocco. J Med Virol 1999. [DOI: 10.1002/(sici)1096-9071(199902)57:2<126::aid-jmv7>3.0.co;2-o] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
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30
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Tsarev SA, Binn LN, Gomatos PJ, Arthur RR, Monier MK, van Cuyck-Gandre H, Longer CF, Innis BL. Phylogenetic analysis of hepatitis E virus isolates from Egypt. J Med Virol 1999; 57:68-74. [PMID: 9890424 DOI: 10.1002/(sici)1096-9071(199901)57:1<68::aid-jmv10>3.0.co;2-e] [Citation(s) in RCA: 54] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
Abstract
Hepatitis E virus (HEV) genome was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in fecal samples of two sporadic cases of hepatitis E in Cairo Egypt. Sequence of the complete putative structural region [open reading frame (ORF)-2] and complete region of unknown function (ORF-3) was determined for the two HEV isolates. Phylogenetic analysis of the nucleotide sequences was performed using neighbor joining or maximum parsimony methods of tree reconstruction. Direct correspondence between the HEV evolutionary trees and geographic origin of the HEV isolates was observed. Three genotypes of HEV were identified: genotype I (Asia-Africa), genotype II (US), and genotype III (Mexico). Genotype I was further divided into two subgenotypes (Asia and Africa). In the Asian subgenotype, three smaller genetic clusters were observed (China-like sequences, Burma-like sequences, and sequence from a fulminant case of HEV). The segregation of all these genetic clusters was supported by the high level of bootstrap probabilities. Four regions of the HEV genome were used for phylogenetic analysis. In all four regions, Egyptian HEV isolates were grouped in a separate African clade.
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Affiliation(s)
- S A Tsarev
- Department of Virus Diseases, Walter Reed Army Institute of Research, Washington, DC 20307, USA.
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Aggarwal R, McCaustland KA, Dilawari JB, Sinha SD, Robertson BH. Genetic variability of hepatitis E virus within and between three epidemics in India. Virus Res 1999; 59:35-48. [PMID: 10854164 DOI: 10.1016/s0168-1702(98)00123-3] [Citation(s) in RCA: 23] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/17/2022]
Abstract
Hepatitis E virus (HEV) is an important cause of epidemic and sporadic acute viral hepatitis in many developing countries, including India. We evaluated the genetic variability within two regions (a 476-nt long ORF1 segment and a 304-nt long ORF2 segment) from specimens collected during three outbreaks in the cities of Karnal (1987), Yamunanagar (1989), and Meerut (1996), India, and from one patient, residing in Lucknow, India, who had a case of sporadic hepatitis (1996). Within an outbreak, sequences in the ORF1 and ORF2 regions were 99.3-100.0% identical. However, when strains were compared between outbreaks, identity in the ORF1 and ORF2 region was 97.1-99.2 and 96.4-100.0%, respectively. A comparison of these sequences to previously published Indian ORF1 and ORF2 sequences revealed even lower similarities, 95.2-98.5 and 95.1-98.7%, respectively. One patient in the Meerut outbreak had genomic sequences that differed substantially from the other patients affected during this outbreak and probably reflected a sporadic infection. The sporadic hepatitis E strain from Lucknow clustered with a previously described HEV strain from a patient with fulminant hepatic failure (FHF). Our data suggest that the ORF1 and ORF2 segments can be used to study the molecular epidemiology of HEV infection and indicate that much remains to be determined about the genetic variability of Indian HEV strains.
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Affiliation(s)
- R Aggarwal
- Hepatitis Branch, Division of Viral and Rickettsial Diseases, National Centerfor Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA
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32
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Meng XJ, Halbur PG, Shapiro MS, Govindarajan S, Bruna JD, Mushahwar IK, Purcell RH, Emerson SU. Genetic and experimental evidence for cross-species infection by swine hepatitis E virus. J Virol 1998; 72:9714-21. [PMID: 9811705 PMCID: PMC110481 DOI: 10.1128/jvi.72.12.9714-9721.1998] [Citation(s) in RCA: 456] [Impact Index Per Article: 16.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/26/1998] [Accepted: 08/24/1998] [Indexed: 12/23/2022] Open
Abstract
Prior to the recent discovery of the swine hepatitis E virus (swine HEV) in pigs from the midwestern United States, HEV was not considered endemic to this country. Since swine HEV is antigenically and genetically related to human strains of HEV, it was important to characterize this new virus further. The infectivity titer of a pool of swine HEV in pigs was determined in order to prepare a standardized reagent and to evaluate the dose response in pigs. Although the sequence of swine HEV varied extensively from those of most human strains of HEV, it was very closely related to the two strains of human HEV (US-1 and US-2) isolated in the United States. The U.S. strains which were recently recovered from two patients with clinical hepatitis E in the United States shared >/=97% amino acid identity with swine HEV in open reading frames 1 and 2. Phylogenetic analyses of different regions of the genome revealed that swine HEV and the U.S. strains grouped together and formed a distinct branch. These results suggested that swine HEV may infect humans. When we inoculated rhesus monkeys and a chimpanzee, experimental surrogates of humans, with swine HEV, the primates became infected. Furthermore, in a reciprocal experiment, specific-pathogen-free pigs were experimentally infected with the US-2 strain of human HEV that is genetically similar to swine HEV. These results provided experimental evidence for cross-species infection by the swine virus. Thus, humans appear to be at risk of infection with swine HEV or closely related viruses.
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Affiliation(s)
- X J Meng
- Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
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33
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Pina S, Jofre J, Emerson SU, Purcell RH, Girones R. Characterization of a strain of infectious hepatitis E virus isolated from sewage in an area where hepatitis E is not endemic. Appl Environ Microbiol 1998; 64:4485-8. [PMID: 9797311 PMCID: PMC106673 DOI: 10.1128/aem.64.11.4485-4488.1998] [Citation(s) in RCA: 97] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Raw sewage samples from an area where hepatitis E is not endemic (Barcelona, Spain) were analyzed by reverse transcriptase-PCR followed by nested PCR. One of the 37 tested samples showed a positive result for hepatitis E virus (HEV). The detected strain was amplified by inoculation into rhesus monkeys, and the course of the infection was studied by analyzing serological and biochemical parameters and by monitoring the presence of HEV in serum and feces. Fecal suspensions from the rhesus monkeys were used as the source of viral particles for sequence analysis. Eighty percent of the genome of the isolated strain, named BCN, was sequenced and found to be phylogenetically related to Asian (Indian) strains, with a 98% nucleotide identity with an isolate from Madras, India. Since this was a single isolation we cannot conclude that HEV is regularly present in the sewage. However, the finding of viable HEV in sewage has implications for contamination of the environment and shellfish by HEV and must be considered in the diagnosis of viral hepatitis in regions of nonendemic hepatitis.
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Affiliation(s)
- S Pina
- Department of Microbiology, Biology School, University of Barcelona, Barcelona, Spain
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Meng J, Pillot J, Dai X, Fields HA, Khudyakov YE. Neutralization of different geographic strains of the hepatitis E virus with anti-hepatitis E virus-positive serum samples obtained from different sources. Virology 1998; 249:316-24. [PMID: 9791023 DOI: 10.1006/viro.1998.9346] [Citation(s) in RCA: 33] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Abstract
A recently developed polymerase chain reaction (PCR)-based cell culture neutralization assay was used to investigate cross-neutralization of known hepatitis E virus (HEV) strains obtained from various HEV-endemic regions of the world with different anti-HEV-positive serum samples. Serum specimens obtained from cynomolgus macaques experimentally infected with strains from Burma, Mexico, or Pakistan cross-neutralized the infectivity of each strain as well as an isolate from Morocco. Serum samples obtained either from infected patients who reside in HEV-endemic regions of the world or from U.S. residents who became infected while traveling to such regions also neutralized all four strains. In contrast, antibodies obtained from rabbits immunized with full-length Burma strain ORF2 protein neutralized only the Burma and Pakistan strains, not the Mexico or Morocco strains. In addition, antibodies obtained from guinea pigs immunized with an N-terminal truncated Burma strain ORF2 protein neutralized each strain except the Morocco strain. These data strongly suggest that antibodies elicited during an HEV infection demonstrate broad HEV neutralizing activity, whereas antibodies elicited after immunization with recombinant Burma ORF2 protein demonstrate a more limited ability to neutralize various HEV strains obtained from different regions of the world endemic for the disease.
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Affiliation(s)
- J Meng
- National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, Georgia, 30333, USA.
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35
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Hsieh SY, Yang PY, Ho YP, Chu CM, Liaw YF. Identification of a novel strain of hepatitis E virus responsible for sporadic acute hepatitis in Taiwan. J Med Virol 1998; 55:300-4. [PMID: 9661839 DOI: 10.1002/(sici)1096-9071(199808)55:4<300::aid-jmv8>3.0.co;2-4] [Citation(s) in RCA: 59] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
Hepatitis caused by the hepatitis E virus (HEV) is a self-limited disease and occurs most frequently as epidemic or sporadic hepatitis in developing countries. The role of HEV in sporadic acute hepatitis in areas without a history of hepatitis E epidemics is obscure. Recently, it was found that more than 10% of the patients with acute non-A, non-B, non-C hepatitis in Taiwan were associated with an acute HEV infection. Nucleotide sequences of the regions within the first open reading frame of HEV were determined in four cases and were 96.7-100% identical to each other. As compared to the isolates from China, Pakistan, Burma, India, Africa, and Mexico, the similarities were, however, only 71.7-79.3%. Phylogenetic analysis revealed that the four Taiwan isolates were categorized as a novel HEV group (the Taiwan strain), which was distinct from all of the strains isolated from other parts of the world. In addition, the isolates from China, Burma, India, and Pakistan were catalogued as the second genotype of HEV (the Asian strain), and the Mexican isolate as the third (the Mexican strain). The African isolate was more related to the Asian type and might be a subtype of the Asian strain. A simple genotyping method by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is described. The findings also support the hypothesis that HEV may be responsible for some sporadic acute non-A, non-B, non-C hepatitis in other developed countries.
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Affiliation(s)
- S Y Hsieh
- Liver Research Unit, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan
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36
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Robinson RA, Burgess WH, Emerson SU, Leibowitz RS, Sosnovtseva SA, Tsarev S, Purcell RH. Structural characterization of recombinant hepatitis E virus ORF2 proteins in baculovirus-infected insect cells. Protein Expr Purif 1998; 12:75-84. [PMID: 9473460 DOI: 10.1006/prep.1997.0817] [Citation(s) in RCA: 110] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
The hepatitis E virus (HEV) capsid antigen has been proposed as a candidate subunit vaccine for the prevention of hepatitis E. The full-length HEV ORF2 protein product is predicted to contain 660 amino acids and to weigh 72,000 daltons. Expression of the HEV ORF2 capsid gene from recombinant baculoviruses in insect cells produced multiple immunoreactive proteins ranging in size from 30 to 100 kDa. The most abundant HEV proteins had molecular weights of 72, 63, 56, and 53 kDa. Temporal expression kinetics of these viral polypeptides indicated that the 72- and 63-kDa polypeptides were produced abundantly within the initial 36 h. postinfection but were replaced by 56- and 53-kDa polypeptides in the cell and medium, respectively, by 48 h postinfection. The 53-kDa protein was secreted as early as 24 h. postinfection, and accumulation in the medium peaked by 72 h postinfection. Purification of the 53-, 56-, and 63-kDa viral polypeptides was accomplished by anion-exchange and subsequent gel filtration chromatography. Sequence analysis of the 53-, 56-, and 63-kDa HEV polypeptides indicated that the amino terminus was amino acid residue 112 of the predicted full-length protein product. The results of carboxy terminal amino acid sequencing indicated that the carboxy terminus of the 53-, 56-, and 63-kDa HEV proteins was located at amino acid residues 578, 607, and 660, respectively. The molecular masses of the 53- and 56-kDa HEV polypeptides were 53,872 and 56,144 as determined by mass spectroscopy.
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Affiliation(s)
- R A Robinson
- Molecular Virology Laboratory, DynCorp, 1 Taft Court, Rockville, Maryland 20850, USA.
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Gouvea V, Hoke CH, Innis BL. Genotyping of hepatitis E virus in clinical specimens by restriction endonuclease analysis. J Virol Methods 1998; 70:71-8. [PMID: 9506814 DOI: 10.1016/s0166-0934(97)00172-9] [Citation(s) in RCA: 19] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
The genomic variability of hepatitis E virus (HEV) was examined by restriction endonuclease analysis (REA) of four genomic cDNA copies comprising a 499 bp segment of the putative polymerase gene, a 264 bp segment of the helicase gene, and two, 680 bp and 448 bp, segments of the capsid gene. Analysis of the deduced restriction sites of all 27 HEV sequences currently available in the GenBank, and digestion of reverse-transcribed and nested PCR amplified segments obtained from six Nepali isolates were used to devise and test a REA genotyping assay. The assay allowed easy discrimination between the Mexico and Asian genotypes, and the classification of the Asian genotypes into three, or perhaps four subgenotypes. In addition, endonucleases identifiers of individual isolate or clusters of isolates were found. This assay permits rapid identification of a large number of HEV isolates directly from clinical specimens for studies on the molecular epidemiology and evolution of HEV.
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Affiliation(s)
- V Gouvea
- Department of Virus Diseases, Walter Reed Army Institute of Research, Washington, DC 20307-5100, USA.
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38
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van Cuyck-Gandré H, Zhang HY, Tsarev SA, Clements NJ, Cohen SJ, Caudill JD, Buisson Y, Coursaget P, Warren RL, Longer CF. Characterization of hepatitis E virus (HEV) from Algeria and Chad by partial genome sequence. J Med Virol 1997; 53:340-7. [PMID: 9407381 DOI: 10.1002/(sici)1096-9071(199712)53:4<340::aid-jmv5>3.0.co;2-7] [Citation(s) in RCA: 59] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
The purpose of this study was to analyze partial nucleotide sequences and derived peptide sequences of hepatitis E virus (HEV) from two outbreaks of hepatitis E in Africa (Chad 1983-1984; Algeria 1978-1980). A portion of ORF3 and the major portion of ORF2 were amplified by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). The PCR products were sequenced directly or after cloning into the pCRII vector. Sequences were then compared to the corresponding regions of reported full length HEV sequences. In the ORF2 and ORF3 regions, the homology between the Algerian and the Chad isolates at the nucleic acid level was 92 and 95%, respectively. At the peptide level the homology was 98% in both regions. In these regions, both strains are more related to Asian strains at the nucleic acid level (89 to 95%) and at the amino acid level (95 to 100%) than to the Mexico strain. At the peptide level the differences are less apparent. Both African isolates have amino acid changes in common with some reference strains although the Chad isolate has three unique changes. These African strains of HEV, based on the ORF2 and ORF3 phylogenetic trees, appear to be a distinct phylogenetic group, separate from the Mexican and Asian strains.
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Affiliation(s)
- H van Cuyck-Gandré
- Department of Virus Diseases, Walter Reed Army Institute of Research, Washington DC, USA
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39
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Gouvea V, Snellings N, Cohen SJ, Warren RL, Myint KS, Shrestha MP, Vaughn DW, Hoke CH, Innis BL. Hepatitis E virus in Nepal: similarities with the Burmese and Indian variants. Virus Res 1997; 52:87-96. [PMID: 9453147 DOI: 10.1016/s0168-1702(97)00112-3] [Citation(s) in RCA: 25] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
Hepatitis E has been the predominant type of acute hepatitis in Nepal both in adults and children, in sporadic and epidemic forms. We examined six hepatitis E virus (HEV) isolates obtained during an 8-year period, from 1987 to 1995, in the Kathmandu valley of Nepal. Analysis of portions of the putative helicase, polymerase and capsid genes demonstrated close genetic relatedness among themselves (> 96.4% identity) and with the Burmese (> 95.5%) and Indian (> 95.3%) isolates, and less so with the African (> 94.4%) and the Chinese (> 91%) isolates within the Asian genotype. Phylogenetic analysis placed the Nepali isolates in the Burma-India evolutionary branch and showed that the oldest isolate, TK78/87 was more similar to the Burmese isolates whereas the most recent isolates were closer to the Indian ones. Assuming no frameshifts, the Nepali isolates showed high amino acid conservation, but also unique changes when compared to other HEV isolates. Amino acid residue 614 of the capsid protein was identified as a possible marker to distinguish the Burma-Nepal-India from the China-Central Asian Republics subgenotype, and the Mexico genotype.
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Affiliation(s)
- V Gouvea
- Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Washington, DC 20307-5100, USA.
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40
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Li TC, Yamakawa Y, Suzuki K, Tatsumi M, Razak MA, Uchida T, Takeda N, Miyamura T. Expression and self-assembly of empty virus-like particles of hepatitis E virus. J Virol 1997; 71:7207-13. [PMID: 9311793 PMCID: PMC192060 DOI: 10.1128/jvi.71.10.7207-7213.1997] [Citation(s) in RCA: 251] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023] Open
Abstract
Hepatitis E virus (HEV) is a pathogenic agent that causes fecally-orally transmitted acute hepatitis. The genome, a single-stranded positive-sense RNA, encodes three forward open reading frames (ORFs), in which an approximately 2-kb structural protein is located in the 3' end. To produce HEV-like particles the structural protein, with its N terminus truncated (amino acid residues 112 to 660 of ORF2), was expressed in insect Tn5 cells by a recombinant baculovirus. In addition to the primary translation product with a molecular mass of 58 kDa, a large amount of a further-processed molecule with a molecular mass of 50 kDa was generated and efficiently released into the culture medium. Electron microscopic observation of the culture medium revealed that the 50-kDa protein self-assembled to form empty virus-like particles (VLPs). The buoyant density of the VLPs in CsCl was 1.285 g/cm3 and their diameter was 23.7 nm, a little smaller than the 27 nm of native HEV particles secreted into the bile or stools of experimentally infected monkeys. The yield of the VLPs was 1 mg per 10(7) cells as a purified form. The particles possess antigenicity similar to that of authentic HEV particles and, consequently, they appear to be a good antigen for the sensitive detection of HEV-specific immunoglobulin G (IgG) and IgM antibodies. Furthermore, the VLP may be the most promising candidate yet for an HEV vaccine, owing to its potent immunogenicity.
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Affiliation(s)
- T C Li
- Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan
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41
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Chatterjee R, Tsarev S, Pillot J, Coursaget P, Emerson SU, Purcell RH. African strains of hepatitis E virus that are distinct from Asian strains. J Med Virol 1997. [DOI: 10.1002/(sici)1096-9071(199710)53:2<139::aid-jmv5>3.0.co;2-a] [Citation(s) in RCA: 50] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
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Meng XJ, Purcell RH, Halbur PG, Lehman JR, Webb DM, Tsareva TS, Haynes JS, Thacker BJ, Emerson SU. A novel virus in swine is closely related to the human hepatitis E virus. Proc Natl Acad Sci U S A 1997; 94:9860-5. [PMID: 9275216 PMCID: PMC23282 DOI: 10.1073/pnas.94.18.9860] [Citation(s) in RCA: 835] [Impact Index Per Article: 29.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023] Open
Abstract
A novel virus, designated swine hepatitis E virus (swine HEV), was identified in pigs. Swine HEV crossreacts with antibody to the human HEV capsid antigen. Swine HEV is a ubiquitous agent and the majority of swine >/=3 months of age in herds from the midwestern United States were seropositive. Young pigs naturally infected by swine HEV were clinically normal but had microscopic evidence of hepatitis, and developed viremia prior to seroconversion. The entire ORFs 2 and 3 were amplified by reverse transcription-PCR from sera of naturally infected pigs. The putative capsid gene (ORF2) of swine HEV shared about 79-80% sequence identity at the nucleotide level and 90-92% identity at the amino acid level with human HEV strains. The small ORF3 of swine HEV had 83-85% nucleotide sequence identity and 77-82% amino acid identity with human HEV strains. Phylogenetic analyses showed that swine HEV is closely related to, but distinct from, human HEV strains. The discovery of swine HEV not only has implications for HEV vaccine development, diagnosis, and biology, but also raises a potential public health concern for zoonosis or xenozoonosis following xenotransplantation with pig organs.
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Affiliation(s)
- X J Meng
- Hepatitis Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 7 Center Drive, MSC 0740, Bethesda, MD 20892, USA.
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Abstract
Hepatitis E has a world-wide distribution and causes substantial morbidity and mortality in some developing countries, particularly among pregnant women. Hepatitis E virus (HEV) has recently been cloned and sequenced, and new diagnostic tests have been developed. These tests have been used to begin to characterize the natural history and epidemiological features of HEV infection. Experimental vaccines have also been developed that offer the potential to prevent hepatitis E. However, much remains to be learned about HEV, including the mechanisms of transmission, the reservoir(s) of the virus, and the natural history of protective immunity in order to develop effective strategies to prevent this disease.
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Affiliation(s)
- E E Mast
- Hepatitis Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA
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44
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Abstract
Hepatitis E has a worldwide distribution and causes substantial morbidity and mortality in some developing countries, particularly among pregnant women. Hepatitis E virus (HEV) has recently been cloned and sequenced and new diagnostic tests have been developed; these tests have been used to begin to characterize the natural history and epidemiologic features of HEV infection. Experimental vaccines have also been developed that offer the potential to prevent hepatitis E. However, to develop effective strategies to prevent this disease, much remains to be learned about HEV, including the vehicles of transmission, the reservoir(s) of the virus, and the natural history of protective immunity.
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Affiliation(s)
- E E Mast
- Hepatitis Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA
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