应用实时荧光RT-PCR法检测水产品中诺如病毒

摘要

Objective To establish a testing method for novovirus in aquatic product samples. Methods Optimized glycine buffer was used to dispose the homogenate of aquatic products, the concentration of concentrated virus in PEG precipitation was tested, virus RNA was extracted, and the method of fluorescent quantitative RT-PCR was applied to test novovirus. Results 72 samples were tested by an optimized method, and positive PCR products were validated by nucleotide sequencing method. The positive PCR products were validated by the nucleotide sequencing method as novovirus. Conclusion The research method can be applied to test novovirus in aquatic products. Few aquatic products in the seafood market in Fushun City are contaminated by novovirus.%目的：建立水产品样品中的诺如病毒检测方法。方法优化甘氨酸缓冲液处理水产品匀浆液，摸索PEG沉淀浓缩病毒的浓度，提取病毒RNA，采用荧光定量RT-PCR方法进行诺如病毒检测。结果应用优化后的方法检测72份样品，并用核苷酸序列测定法对阳性PCR产物进行验证，所得阳性PCR产物经核苷酸序列测序证实为诺如病毒。结论本研究方法可以应用于水产品中的诺如病毒检测，抚顺市海鲜市场上的水产品很少存在诺如病毒的污染。