Establishment and application of TaqMan real-time RT-PCR for the detection of hepatitis E virus.
- Author:
Qing-Ling MENG
1
;
Feng QIU
;
Li-Ping SHEN
;
Sheng-Li BI
Author Information
- Publication Type:Journal Article
- MeSH: DNA Primers; genetics; Hepatitis E; virology; Hepatitis E virus; genetics; isolation & purification; Humans; RNA, Viral; genetics; metabolism; Real-Time Polymerase Chain Reaction; methods; Taq Polymerase; metabolism
- From: Chinese Journal of Experimental and Clinical Virology 2012;26(6):486-488
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a specific TaqMan-based Real-time PCR assay for the detection of hepatitis E virus (HEV).
METHODSAccording to the references, primers-probe sets which were located in ORF2, the conservative part of HEV genome were designed and therefore we established a HEV TaqMan real-time RT-PCR assay with great performance of specificity, sensitivity and reproducibility. And then it was used in the detection of HEV RNA in clinical samples.
RESULTSThe HEV Real-time RT-PCR assay established in this study were able to detect HEV RNA with a detection limit of 10 copies/reaction. When the detection of a same sample was repeated for several times, coefficients of variation (CV) was all less than 1.53%. Our data also suggested that there were 1.87 x 10(6)-8.12 x 10(9) RNA copies in 1 ml of the clinical samples.
CONCLUSIONThe TaqMan-based Real-time PCR assay established in this study was specific and precise for the rapid detection of HEV RNA. It was applied successfully in the pathogen detection of clinical samples.
