Comprehensive Molecular Approach for Characterization of Hepatitis E Virus Genotype 3 Variants
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Comprehensive Molecular Approach for Characterization of Hepatitis E Virus Genotype 3 Variants. / Wang, Bo; Harms, Dominik; Papp, C Patrick; Niendorf, Sandra; Jacobsen, Sonja; Lütgehetmann, Marc; Pischke, Sven; Wedermeyer, Heiner; Hofmann, Jörg; Bock, C-Thomas.
in: J CLIN MICROBIOL, Jahrgang 56, Nr. 5, 05.2018, S. e01686-17.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Comprehensive Molecular Approach for Characterization of Hepatitis E Virus Genotype 3 Variants
AU - Wang, Bo
AU - Harms, Dominik
AU - Papp, C Patrick
AU - Niendorf, Sandra
AU - Jacobsen, Sonja
AU - Lütgehetmann, Marc
AU - Pischke, Sven
AU - Wedermeyer, Heiner
AU - Hofmann, Jörg
AU - Bock, C-Thomas
N1 - Copyright © 2018 American Society for Microbiology.
PY - 2018/5
Y1 - 2018/5
N2 - Autochthonous hepatitis E virus genotype 3 (HEV-3) infections in industrialized countries are more frequent than previously assumed. HEV-3 is zoonotic and the causal pathogen of chronic hepatitis E. According to the latest classification of the family Hepeviridae, 10 designated HEV-3 subtypes (HEV-3a to HEV-3j) and 7 unassigned HEV-3 subtypes are proposed. In order to identify and characterize the HEV-3 variants in circulation, we developed a molecular approach combining a sensitive HEV-specific real-time reverse transcription-PCR (RT-PCR) targeting the overlapping region of HEV ORF2 and ORF3 (the ORF2/3 region) and two newly designed consensus nested RT-PCRs targeting the HEV ORF1 and ORF2 genes, respectively. Since complete genome sequences are required for new HEV-3 subtype assignment, we implemented a straightforward approach for full-length HEV-3 genome amplification. Twenty-nine human serum samples and six human feces samples from chronic hepatitis E patients were selected for evaluation of the system. Viral loads ranged from 1 × 104 to 1.9 × 1010 copies/ml of serum and from 1.8 × 104 to 1 × 1012 copies/g of feces. Sequence and phylogenetic analyses of partial ORF1 and ORF2 sequences showed that HEV strains had considerable genetic diversity and clustered into the HEV-3c (29/35), HEV-3e (2/35), HEV-3f (2/35), and unassigned HEV-3 (2/35) subtypes. Moreover, from these strains, three full-length HEV-3 genome sequences were generated and characterized. DE/15-0030 represents a typical HEV-3c strain (95.7% nucleotide identity to wbGER27), while DE/15-0031 and SW/16-0282 have <89.2% homology to known HEV-3 strains and are phylogenetically divergent, indicating novel HEV-3 subtypes. In summary, our approach will significantly facilitate the detection, quantification, and determination of HEV-3 strains and will thus help to improve molecular diagnostics and our knowledge of HEV diversity and evolution.
AB - Autochthonous hepatitis E virus genotype 3 (HEV-3) infections in industrialized countries are more frequent than previously assumed. HEV-3 is zoonotic and the causal pathogen of chronic hepatitis E. According to the latest classification of the family Hepeviridae, 10 designated HEV-3 subtypes (HEV-3a to HEV-3j) and 7 unassigned HEV-3 subtypes are proposed. In order to identify and characterize the HEV-3 variants in circulation, we developed a molecular approach combining a sensitive HEV-specific real-time reverse transcription-PCR (RT-PCR) targeting the overlapping region of HEV ORF2 and ORF3 (the ORF2/3 region) and two newly designed consensus nested RT-PCRs targeting the HEV ORF1 and ORF2 genes, respectively. Since complete genome sequences are required for new HEV-3 subtype assignment, we implemented a straightforward approach for full-length HEV-3 genome amplification. Twenty-nine human serum samples and six human feces samples from chronic hepatitis E patients were selected for evaluation of the system. Viral loads ranged from 1 × 104 to 1.9 × 1010 copies/ml of serum and from 1.8 × 104 to 1 × 1012 copies/g of feces. Sequence and phylogenetic analyses of partial ORF1 and ORF2 sequences showed that HEV strains had considerable genetic diversity and clustered into the HEV-3c (29/35), HEV-3e (2/35), HEV-3f (2/35), and unassigned HEV-3 (2/35) subtypes. Moreover, from these strains, three full-length HEV-3 genome sequences were generated and characterized. DE/15-0030 represents a typical HEV-3c strain (95.7% nucleotide identity to wbGER27), while DE/15-0031 and SW/16-0282 have <89.2% homology to known HEV-3 strains and are phylogenetically divergent, indicating novel HEV-3 subtypes. In summary, our approach will significantly facilitate the detection, quantification, and determination of HEV-3 strains and will thus help to improve molecular diagnostics and our knowledge of HEV diversity and evolution.
KW - Journal Article
U2 - 10.1128/JCM.01686-17
DO - 10.1128/JCM.01686-17
M3 - SCORING: Journal article
C2 - 29514938
VL - 56
SP - e01686-17
JO - J CLIN MICROBIOL
JF - J CLIN MICROBIOL
SN - 0095-1137
IS - 5
ER -
