基于受体捕获诺如病毒检测方法的特异性及应用

Specificity and application of a norovirus detection method based on receptor capturing

目的评价新型感染性诺如病毒检测方法原位捕获反转录实时定量PCR（In situ capture realtime quantitative reversetranscription polymerase chain reaction，ISC-RT-qPCR）的特异性，并将其应用于新鲜草莓中诺如病毒的检测。方法以诺如病毒GⅡ组不同基因型和不同浓度的临床粪便标本评价ISC-RT-qPCR方法的特异性。此外，利用ISC-RT-qPCR方法对以GⅡ组诺如病毒人工污染新鲜草莓样本进行检测。结果ISC-RT-qPCR可特异性检测所有GⅡ诺如病毒8种基因型临床腹泻样本；与传统RT-qPCR相比，检测的Ct值并未随着诺如病毒滴度的降低而升高。人工污染新鲜草莓的ISC-RT-qPCR检测限为1.36基因组拷贝数。结论ISC-RT-qPCR是一种可用于临床腹泻样本和草莓中GⅡ组感染性诺如病毒的检测方法，为食品中诺如病毒的检测与监测提供了技术储备。

ObjectiveTo evaluate of the specificity of a new norovirus （NoV） detection method of in situ capture real-time quantitative reverse transcription polymerase chain reaction （ISC-RT-qPCR） and to apply the method for the detection of NoVs in fresh strawberry.MethodsA panel of stool samples with different NoV genotypes and various inoculums were used for the experiments.ResultsWe found that all the tested samples of eight genogroup Ⅱ （GⅡ） NoVs could be detected specifically by ISC-RT-qPCR. Moreover, in contrast to the conventional RT-qPCR method , the situation that the Ct value increased as the inoculum of NoV GⅡ decreased was not shown using ISC-RT-qPCR. When we tested NoVs in strawberry samples by ISC-RT-qPCR, the minimum test limit could reach 1.36 genocopy/10 g of fresh strawberry.ConclusionsISC-RT-qPCR is an effective and specific technic and it could be applied for the detection of infectious NoVs from stool samples and fresh strawberry samples.