鱼肉制品中巴沙鱼源性成分的实时荧光聚合酶链式反应快速检测法

Real-time polymerase chain reaction method for rapid detection of Pangasius bocourti-derived components in fish products

目的建立巴沙鱼源性成分的实时荧光聚合酶链式反应(PCR)快速检测手段。方法根据巴沙鱼的线粒体cytb基因序列设计引物,使用实时荧光PCR进行扩增,从而达到快速检测产物的目的。结果此方法特异性良好,巴沙鱼基因组DNA灵敏度可达到10^(-4) ng,在与婴幼儿米粉、儿童副食芝麻粉、鸡肉粉和大西洋鳕鱼粉混合的鱼肉制品中均可检测,且质量分数灵敏度均可达到0.01%。结论本方法检测特异性高、时间短、灵敏度高,可满足鱼肉制品中巴沙鱼掺假的检测要求。

Objective To establish a real-time polymerase chain reaction(PCR)rapid detection method for Pangasius bocourti-derived components.Methods Primers were designed according to the mitochondrial cytb gene sequence of Pangasius bocourti,and real-time PCR was used for amplification to achieve the purpose of rapid detection of products.Results This method had good specificity,and the sensitivity could reach 10^(-4) ng Pangasius bocourti DNA.Pangasius bocourti cytb gene could be detected in fish products mixed with rice noodles,sesame powder,chicken powder and atlantic cod meal,and the sensitivity could reach 0.01%.Conclusion This method had high specificity,short time and high sensitivity,and could meet the detection requirements of Pangasius bocourti adulteration in fish meat products.