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

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2012 Dec;26(6):486-8.
[Article in 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 x 10(6)-8.12 x 10(9) RNA copies in 1 ml 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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Primers / genetics
  • Hepatitis E / virology*
  • Hepatitis E virus / genetics
  • Hepatitis E virus / isolation & purification*
  • Humans
  • RNA, Viral / genetics
  • RNA, Viral / metabolism
  • Real-Time Polymerase Chain Reaction / methods*
  • Taq Polymerase / metabolism

Substances

  • DNA Primers
  • RNA, Viral
  • Taq Polymerase