常规RT-PCR与荧光定量RT-PCR检测GⅠ、GⅡ型诺如病毒方法的研究和应用

Research and Application the Conventional RT-PCR and Fluorescence Quantitative RT-PCR in Detection of GⅠ,GⅡType of Norovirus

目的研究常规RT-PCR与荧光定量RT-PCR检测GⅠ、GⅡ型诺如病毒,比较两种检测方式的效果。方法选取2019年7月至2020年7月我院收治的210例腹泻患者粪便为研究对象,应用常规RT-PCR和荧光定量RT-PCR进行检测,比较两种检测方法的检测结果。结果两种检测方法特异性均较强,与轮状病毒、札幌病毒、星状病毒之间无交叉反应,相同体系GⅠ、GⅡ型诺如病毒之间无干扰。常规RT-PCR的最低检测下限是103 copies/μL,荧光定量RT-PCR的最低检测下限是102 copies/μL。常规RT-PCR的检测率为5.71%(12/210),符合率为66.67%(12/18),荧光定量RT-PCR的检测率为8.57%(18/210),符合率为100.00%(18/18);对18例阳性样本进行测序分析,均证实为诺如病毒。结论常规RT-PCR与荧光定量RT-PCR均能快速检测诺如病毒,荧光定量RT-PCR更加灵敏,具有较高的符合率。

Objective To study the conventional RT-PCR and fluorescence quantitative RT-PCR in detection of GⅠ,GⅡtype of norovirus,compare the effects of the two detection methods.Methods The feces of 210 patients with diarrhea from July 2019 to July 2020 were selected as the research object,conventional RT-PCR and fluorescence quantitative RT-PCR were used for detection,the test results of the two methods were compared.Results These two tests were highly specific,there was no cross-reaction with rotavirus,sapporo virus and stavirus,there was no interference between GⅠand GⅡnorovirus in the same system.The minimum detection limit of conventional RT-PCR was 103 copies/μL,the minimum detection limit of fluorescence quantitative RTPCR was 102 copies/μL.The detection rate of conventional RT-PCR was 5.71%(12/210),the coincidence rate was 66.67%(12/18),the detection rate of fluorescence quantitative RT-PCR was 8.57%(18/210),the coincidence rate was 100.00%(18/18);18 positive samples were sequenced and analyzed,both were confirmed as norovirus.Conclusion Both conventional RT-PCR and fluorescence quantitative RT-PCR can detect norovirus rapidly,fluorescence quantitative RT-PCR was more sensitive,it has a high coincidence rate.