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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.

Citation

Dong-Mei Zhou, Miao Jin, Hui-Ying Li, Zi-Qian Xu, Zhao-Jun Duan. Establish A duplex fluorescent quantitative one-step RT-PCR system for the detection of norovirus genogroup I and II]. Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui]. 2013 May;29(3):310-5

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PMID: 23905476

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