贻贝中诺如病毒荧光定量反转录PCR检测及北京市污染情况调查

The detection of Norovirus in mussels sold in Beijing by using TaqMan-based one-step quantitative RT-PCR assays

目的监测北京市市售贻贝中诺如病毒(NoV)的污染率及污染浓度,为贝类样品中诺如病毒的风险评估提供基础数据。方法解剖并分离贻贝的内脏团,将内脏团研磨至均质,加入磷酸盐缓冲液振荡60 min,提取病毒颗粒。提取病毒RNA,荧光定量反转录PCR检测诺如病毒,并进行定量分析。结果共检测293份贻贝样品,总阳性率为9.22%(27/293),其中诺如病毒基因组Ⅰ(GGⅠ)占阳性样品数的37.04%(10/27),基因组Ⅱ(GGⅡ)占阳性样品数的62.96%(17/27),未检测到同时携带GGⅠ和GGⅡ的样品。阳性样品中诺如病毒浓度范围在6.20×10~3~3.15×10~5基因拷贝/g(内脏)之间。结论北京市市售贻贝中存在诺如病毒污染。

Objective To monitor the contamination rate and concentration of Norovirus in mussels sold in Beijing seafood market and to provide basic data for risk assessment of Norovirus in mussels. Methods The visceral mass of mussels were dissected and homogenized,Norovirus were extracted from visceral mass by adding PBS buffer and shaking 60 min. The Norovirus RNA was extracted and detected using commercial kit. Results Two hundred and ninety-three mussels samples were detected. The total positive rate was 9. 22%（ 27/293）. In all the positive samples,the gene group Ⅰ accounted for 37. 04%（ 10/27） and gene group Ⅱ accounted for 62. 96%（ 17/27）. The range of the quantity of Norovirus in positive mussels was 6. 20 × 10 3-3. 15 × 10 5 gene copies/g（ visceral mass）. Conclusion The mussels sold in Beijing were contaminated by Norovirus.