title: [Establish A duplex fluorescent quantitative one-step RT-PCR system for the detection of norovirus genogroup I and II].
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.
authors.affiliation: National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. zhoudongmeicdc@163.com
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Zhou
authors.forename: Miao
authors.initials: M
authors.lastname: Jin
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Li
authors.forename: Zi-Qian
authors.initials: ZQ
authors.lastname: Xu
authors.forename: Zhao-Jun
authors.initials: ZJ
authors.lastname: Duan
mesh_terms: D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000821:virology; D005759:Gastroenteritis / Q000175:diagnosis / Q000821:virology*; D005838:Genotype; D006801:Humans; D029322:Norovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
chemical_list: D017931:DNA Primers