Validation of single real-time TaqMan® PCR assay for the detection and quantitation of four major genotypes of hepatitis E virus in clinical specimens

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 × 101 to 7.2 × 103 RNA copies/µl in clinical samples (serum and stools). J. Med. Virol. 78:1076-1082, 2006. © 2006 Wiley-Liss, Inc.