Establishment and application of TaqMan real-time RT-PCR for the detection of hepatitis E virus

VernacularTitle:戊型肝炎病毒TaqMan Real-time RT-PCR法的建立及应用
Author: Qing-Ling MENG ¹ ; Feng QIU ; Li-Ping SHEN ; Sheng-Li BI
Author Information

1. 中国疾病预防控制中心病毒病预防控制所
Keywords: Hepatitis E virus; Reverse transcriptase polymerase chain reaction; Diagnosis
From: Chinese Journal of Experimental and Clinical Virology 2012;26(6):486-488
CountryChina
Language:Chinese
Abstract: Objective To establish a specific TaqMan-based Real-time PCR assay for the detection of hepatitis E virus (HEV).Methods According 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.Results The 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 × 106-8.12 × 109 RNA copies in 1ml of the clinical samples.Conclusion The 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.