Establishment and application of TaqMan real-time RT-PCR for the detection of hepatitis A virus
10.3760/cma.j.issn.1003-9279.2012.02.022
- VernacularTitle:甲肝病毒特异性TaqMan荧光定量PCR法检测甲肝病毒核酸的研究
- Author:
Hui-Hui ZHENG
1
;
Feng QIU
;
Jing-Yuan CAO
;
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 A virus;
Reverse traseriptase polymerace chain reaction;
Sensitivity and specificity
- From:
Chinese Journal of Experimental and Clinical Virology
2012;26(2):142-144
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a specific TaqMan-based Real-time PCR assay for the detection of hepatitis A virus in serum samples.Methods According to the references,primers-probe sets which were located in 5'-NCR,the most conservative part of HAV genome were designed and therefore we established a TaqMan real-time RT-PCR assay with great performance of specificity, sensitivity and reproducibility.And then it was used in the detection of HAV RNA in serum from HAV patients.Results The HAV Real-time RT-PCR assay established in this study were able to detect HAV RNA and its detection limit ranged from 0.1CCID50/reaction to 0.01CCID50/reaction.When the detection of a same sample was repeated for three times,coefficients of varistion (CV) of intra- and inter-assay were calculated and they were all less than 2.0％ and 2.6％ respectively.Our data suggested that there were 5.18 × 102 - 4.93 × 107 RNA copies in 1 ml of the serum from acute HAV patients.Conclusion The TaqMan-based Real-time PCR assay established in this study was specific and precise for the rapid detection of HAV RNA.It was applied successfully in the pathogen detection of clinical samples.
