二重实时荧光PCR方法检测山羊和绵羊源性成分

Differentiation of goat and sheep-derived materials by fluorescent real-time PCR

本研究根据山羊、绵羊线粒体中的cytb基因序列,利用ABI primer express 3.0和DNAStar软件设计了特异性引物和探针。通过对引物和探针浓度、Mg2+浓度、dNTP浓度和Taq酶用量以及反应条件等因素的优化筛选,建立了能同时鉴别山羊和绵羊源性成分的二重实时荧光PCR方法。所建方法灵敏性高、特异性好,对17种不同源性的动物DNA和50份不同来源的样品DNA进行实时荧光PCR检测,结果表明引物与探针能特异地鉴别检测出山羊和绵羊源性成分。实验结果表明该检测方法适用于饲料、肉制品、奶制品和生皮等动物源性产品的山羊和绵羊源性成分鉴定。

In this study, a real time PCR assay capable of differentiating goat and sheep-derived materials was developed using specific primers and probes designed according to goat and sheep mitochondrial DNA cyt b gene sequences. This method was 10 times as sensitive as normal PCR, and successfully detected goat and sheep-derived materials in 50 samples of 17 animals from different sources. Therefore this method has the potential application of detection for goat and sheep-derived materials in feed, meat, milk and pelt, etc.