牡蛎中诺如病毒核酸两种提取方法比较

摘要

目的 寻找简便有效的更适合长期监测的牡蛎中诺如病毒核酸提取的方法.方法 通过甘氨酸聚乙二醇富集后硅胶膜离心柱法(方法A)和蛋白酶K生物消化后顺磁性硅胶基法(方法B)两种核酸提取方法,采用实时RT-PCR检测自然状态的牡蛎和人工染毒状态的牡蛎来进行比较.结果 两种方法的阳性检出率没有差异,总体比较方法B的病毒回收率高.结论 方法B优于方法A,且方法B更快捷,故方法B更适合长期监测.%Objective To explore a convenient and effective method for norovirus nucleic acid extraction from oysters suitable for long-term viral surveillance. Method Two methods, namely method A (glycine washing and polyethylene glycol precipitation of the virus followed by silica gel centrifugal column) and method B (protease K digestion followed by application of paramagnetic silicon) were compared for their performance in norovirus nucleic acid extraction from oysters. Real-time RT-PCR was used to detect norovirus in naturally infected oysters and in oysters with induced infection. Results The two methods yielded comparable positive detection rates for the samples, but the recovery rate of the virus was higher with method B than with method A. Conclusion Method B is a more convenient and rapid method for norovirus nucleic acid extraction from oysters and suitable for long-term surveillance of norovirus.