贝类中诺如病毒实时荧光定量PCR检测方法的建立及应用

Establishment and application of RT-PCR in the detection of norovirus in shellfish

目的建立实时荧光定量PCR法检测GⅠ、GⅡ型诺如病毒的方法。对东海区域市场内零售贝类海产品中诺如病毒携带情况进行监测,为海产品食品安全和风险预警提供科学依据。方法采集浙江沿海地区市场上常见贝类,利用实时荧光定量PCR检测技术对贝类进行检测。结果 2 955份样品中有63份样品携带诺如病毒,其中杂色蛤、太平洋牡蛎、僧帽牡蛎、毛蚶中均检出GⅡ型诺如病毒。贝类样品中太平洋牡蛎的检出率最高,为15.64%。贝类中诺如病毒在冬季（11月-次年2月）的检出率明显高于夏季（6月-9月）,其季节分布差异有统计学意义（χ^2=31.3,P〈0.05）。结论东海区域贝类中,特别是生食或半熟食贝类中诺如病毒的携带水平较高、分布较广,应持续做好贝类中诺如病毒携带监测,同时做好风险预警。

Objective To establish the method of detecting GI and GII norovirus through real-time RT-PCR,so as to provide scientific basis for seafood food safety and early risk warning through the detection of the norovirus in retail shellfish seafood in the area of East China Sea.Methods Common shellfish on the market of Zhejiang coastal areas was collected and then detected through RT-PCR detection technology.Results Among the 2 955 samples,63 samples carried norovirus.To be specific,GII norovirus was detected in venerupis variegata,crassostrea gigas,ostrea cucullata and scapharca subcrenata.Among the shellfish samples,crassostrea gigas had the highest detection rate of norovirus（15.64%）.Besides,the detection rate of norovirus in shellfish in winter（November to February） was significantly higher than that in summer（June to September）,with the season distribution difference statistically significant（χ^2= 31.3,P〈0.05）.Conclusion Shellfish in the East China sea area,especially raw or half cooked shellfish,had high incidence and wide distribution of norovirus.Thus,it is advisable to constantly monitor norovirus carrying in shellfish and do early risk warning measure.