一步法多重实时 RT-PCR 检测方法用于快速同时检测人诺如病毒基因 I、II 和 IV 型。
A one-step multiplex real-time RT-PCR assay for rapid and simultaneous detection of human norovirus genogroup I, II and IV.
机构信息
Jiaxing Center for Disease Control and Prevention, Wen Qiao Road 486, Jiaxing 314050, China.
出版信息
J Virol Methods. 2013 May;189(2):277-82. doi: 10.1016/j.jviromet.2013.02.004. Epub 2013 Feb 27.
A one-step multiplex real-time reverse transcription-PCR (RT-PCR) assay was developed for one-tube and simultaneous detection of three genogroups of human norovirus, genogroup I, II and IV (GI, GII and GIV). The specificity and sensitivity of the assay were evaluated and 50 samples were tested by using this assay. The results showed that the multiplex assay had high sensitivity and specificity. The amplification efficiencies of the assay were 91.3%, 90.1%, 88.9% and the detection limits were up to 16.9, 6.3, 43.0 copies/reaction respectively for norovirus GI, GII and GIV detection. No cross-reaction with the other examined RNA viruses was observed, and the qualitative analysis of samples showed that the multiplex assay had a good consistency with its corresponding monoplex assays for the detection of norovirus GI, GII and GIV (Kappa values were 0.848, 0.876 and 0.812 respectively).
建立了一种一步法多重实时逆转录聚合酶链反应（RT-PCR）检测方法，用于在一管中同时检测三种人类诺如病毒基因型，即基因型 I、II 和 IV（GI、GII 和 GIV）。评估了该检测方法的特异性和灵敏度，并使用该方法检测了 50 个样本。结果表明，该多重检测方法具有较高的灵敏度和特异性。该检测方法的扩增效率分别为 91.3%、90.1%和 88.9%，检测限分别达到 16.9、6.3 和 43.0 拷贝/反应，用于检测诺如病毒 GI、GII 和 GIV。与其他检测的 RNA 病毒无交叉反应，对样本的定性分析表明，该多重检测方法与相应的单重检测方法检测诺如病毒 GI、GII 和 GIV 的一致性良好（kappa 值分别为 0.848、0.876 和 0.812）。
Zotero 插件