常见肉类中鸭源性成分荧光PCR检测方法的建立

目的通过筛选引物、探针体系,优化PCR反应条件,建立常见肉品中鸭源性DNA的荧光PCR检测方法。方法针对线粒体基因设计50对引物探针,以鸭DNA为模板,利用PCR和荧光PCR方法筛选引物探针并确定反应条件,进行灵敏度、特异性和稳定性试验,最终确定荧光定量PCR方法的引物、探针及反应条件。结果当样本含量为0.1%时,本方法仍可特异、灵敏地检出鸭肉成分,检测Ct值可稳定在20以下。结论建立了常见肉品中鸭源性DNA的荧光PCR检测方法,可用于肉类制品中鸭源性成分的鉴定。

Establishment of real-time fluorescence quantitative PCR method for identi-fying duck ingredients from edible meat

Objective To establish a real-time fluorescence quantitative PCR method for identifying the hidden duck ingredients in other meats through screening of primers and probe system and optimizing of PCR reaction conditions. Methods Totally fifty primer pairs and probes were designed according to sequence of the cytochrome b gene from the duck. Primers and probe were selected and the conditions of the PCR were confirmed. Specificity and sensitivity of the method were evaluated. Results Duck ingredients could be identified specifically and sensitively, even the content percentage was 0.1%. The Ct value was below 20. Conclusion The real-time fluorescence quantitative PCR method for identification of duck has high specificity and sensitivity. This method can be used for identifying hidden duck ingredients in other meats.