title: Validation of single real-time TaqMan PCR assay for the detection and quantitation of four major genotypes of hepatitis E virus in clinical specimens.
abstract: 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 x 10(1) to 7.2 x 10(3) RNA copies/microl in clinical samples (serum and stools).
authors.affiliation: National Reference Centre for Hepatitis E, Teaching Military Hospital Val de Grace, Paris, France. vincent.enouf@Libertysurf.fr
authors.forename: V
authors.initials: V
authors.lastname: Enouf
authors.forename: G
authors.initials: G
authors.lastname: Dos Reis
authors.forename: J P
authors.initials: JP
authors.lastname: Guthmann
authors.forename: P J
authors.initials: PJ
authors.lastname: Guerin
authors.forename: M
authors.initials: M
authors.lastname: Caron
authors.forename: V
authors.initials: V
authors.lastname: Marechal
authors.forename: E
authors.initials: E
authors.lastname: Nicand
mesh_terms: D016679:Genome, Viral; D005838:Genotype; D016751:Hepatitis E / Q000821:virology*; D016752:Hepatitis E virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D019914:Taq Polymerase / Q000378:metabolism*
chemical_list: D019914:Taq Polymerase