Abstract:To establish a TaqMan-based real-time fluorescent quantitative PCR assay for detection and quantitation of Hepatitis E virus. The conserved region ORF2 was used for the design of primers and TaqMan probe based on the alignment result of HEV sequences downloaded from Genbank. The sensitivity, stability and specificity of the established method was evaluated. It was demonstrated that the TaqMan assay and detect as few as 5 genome equivalent (GE) copies of HEV plasmid DNA per reaction and showed a high linear dynamic range of quantitation(5×100-5×108 molecules of copies/reaction) with a good correlation. The coefficient of variation(CV)of intra-assay for same sample was 1.49%(n=10) and those of inter-assay were 1.98% and 2.23%(n=5). The specificity was confirmed by detecting 22 human faecal samples. This Taqman based real-time PCR assay is sensitive, stable and specific for detection of HEV.
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