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Validation of single real‐time TaqMan® PCR assay for the detection and quantitation of four major genotypes of hepatitis E virus in clinical specimens
Author(s) -
Enouf V.,
Dos Reis G.,
Guthmann J.P.,
Guerin P.J.,
Caron M.,
Marechal V.,
Nicand E.
Publication year - 2006
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.20665
Subject(s) - taqman , hepatitis e virus , virology , real time polymerase chain reaction , biology , genotype , caliciviridae , microbiology and biotechnology , virus , norovirus , gene , genetics
Since the characterization of the genome of the hepatitis E virus (HEV) in 1990, a large genetic diversity has been described. A single real‐time reverse transcription (RT)‐PCR assay with TaqMan® technology has been validated which uses only one set of primers and probe within the ORF2 HEV region (nt 5207–5292) for the detection and quantification of the four major genotypes of HEV. This assay proved to be as efficient as the conventional RT‐PCR methodology for the detection of HEV in clinical samples testing positive previously. The real‐time RT‐PCR and conventional RT‐PCR were performed comparatively on 60 pairs of sera and stools collected during a recent outbreak of hepatitis E in Darfur. The real‐time RT‐PCR assay was 10‐ to 100‐fold sensitive than for conventional RT‐PCR assays used in this study with a range quantitation from 1.8 × 10 1 to 7.2 × 10 3 RNA copies/µl in clinical samples (serum and stools). J. Med. Virol. 78:1076–1082, 2006. © 2006 Wiley‐Liss, Inc.