双重实时荧光PCR法鉴别大西洋鲑鱼和虹鳟鱼

Simultaneous identification of Salmo salar and Oncorhynchus mykiss by dual real-time PCR

目的建立双重实时荧光PCR法同时鉴别大西洋鲑鱼和虹鳟鱼的检测方法。方法在现有单一物种检测方法的基础上,应用多重实时荧光PCR检测技术对大西洋鲑鱼和虹鳟鱼同时进行检测,并对PCR反应时间和反应温度进行优化。结果该方法通过1管PCR反应实现对大西洋鲑鱼和虹鳟鱼的同时鉴别,反应时间缩短至1 h以内,方法的特异性好,灵敏度可达到0.1%(DNA质量分数)。结论该方法检测效果好、时间短、成本低,可为监管部门解决三文鱼市场监管问题提供技术手段。

Objective To establish a method for the simultaneous identification of Salmo salar and Oncorhynchus Mykiss by dual real-time PCR.Methods On the basis of the reported single species detection method, multiple real-time PCR technology was applied to simultaneous identification of Salmo salar and Oncorhynchus mykiss, and the PCR reaction time and reaction temperature were optimized.Results The method could identify Salmo salar and Oncorhynchus mykiss by one PCR reaction, and the reaction time was shortened to less than 1 h.The specificity was well, and the sensitivity was up to 0.1%(DNA mass percent).Conclusion This method has the advantages of good detection effect, short operation time and low detection cost, which can provide technical support for the supervision department to solve problems of salmon market.