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目的建立果蔬样品表面诺如病毒快速有效的检测方法,评价酸吸附-碱洗脱-PEG沉淀法对果蔬样品表面诺如病毒分离富集的适用性,为诺如病毒疫情的控制和食源性疾病的溯源提供可靠的实验室依据。方法在北京地区采集草莓样品,采用酸吸附-碱洗脱-PEG方法分离富集样品表面病毒颗粒,设计GⅠ和GⅡ的特异性引物和探针,以实时定量RT-PCR法检测病毒核酸。同时优化病毒提取方法,以Mengo病毒做为过程控制对照,制作不同浓度的标准曲线,计算病毒回收率。结果酸吸附-碱洗脱-PEG法病毒回收率为2.36%,符合病毒检测回收率的要求,与其他食源性病毒没有交叉反应,能够应用于日常检测。结论酸吸附-碱洗脱-PEG方法病毒回收率较高,灵敏度和重复性较好,进一步优化后可以应用于果蔬样品的日常检测。
OBJECTIVE To establish a rapid and effective method for the detection of Norovirus on the surface of fruits and vegetables and to evaluate the applicability of acid-base elution-PEG precipitation on the separation and enrichment of Norovirus on the surface of fruits and vegetables, to control Norovirus epidemic and foodborne diseases Traceability provides a reliable laboratory basis. Methods Strawberry samples were collected in Beijing area. The virus particles were separated and enriched on the surface of the samples by acid-adsorption-alkaline elution-PEG method. The specific primers and probes of GⅠ and GⅡ were designed and used to detect the viral nucleic acid by real-time RT-PCR. At the same time optimize the virus extraction method to Mengo virus as a process control, the production of different concentrations of the standard curve, calculate the virus recovery. Results The recovery rate of acid-adsorption-alkaline elution-PEG method was 2.36%, which was in line with the detection rate of virus recovery. It did not cross-react with other food-borne viruses and could be applied to routine testing. Conclusion The acid-adsorption-alkaline elution-PEG method has high virus recovery rate, good sensitivity and repeatability, and can be further applied to routine testing of fruits and vegetables samples after further optimization.