Establish A duplex fluorescent quantitative one-step RT-PCR system for the detection of norovirus genogroup I and II.
- Author:
Dong-Mei ZHOU
1
;
Miao JIN
;
Hui-Ying LI
;
Zi-Qian XU
;
Zhao-Jun DUAN
Author Information
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. zhoudongmeicdc@163.com
- Publication Type:Journal Article
- MeSH:
DNA Primers;
genetics;
Feces;
virology;
Gastroenteritis;
diagnosis;
virology;
Genotype;
Humans;
Norovirus;
classification;
genetics;
isolation & purification;
Reverse Transcriptase Polymerase Chain Reaction;
instrumentation;
methods
- From:
Chinese Journal of Virology
2013;29(3):310-315
- CountryChina
- Language:Chinese
-
Abstract:
The object of this study is to develop a duplex fluorescent quantitative one-step RT-PCR assay for detection and quantitation of GI and GII norovirus. The specific primers, Taqman probes, optimized reaction solution and condition were used to develop the duplex fluorescent quantitative one-step RT-PCR assay. The sensitivity, specificity and reproducibility of the assay were evaluated. The assay was evaluated by testing the 100 specimen samples and compared with the reference assay conventional RT-PCR. The assay possessed high specificity for norovirus detection without any evident cross-reaction with enteric adenovirus, rotavirus or astrovirus. The detection limit of the real-time RT-PCR assay, for GI and GII norovirus was up to 10(3) copy/microL respectively. Compared with the conventional RT-PCR assay, the assay in this study had higher sensitivity with higher detection rate of norovirus in stool specimens. The duplex fluorescent quantitative one-step RT-PCR assay provides rapid, sensitive and reliable detection of GI and GII norovirus, and could be used as a laboratory diagnosis of norovirus in acute gastroenteritis patients.
