牛肉及其制品中肉类掺假的荧光PCR鉴别体系优化

Optimization of real-time PCR identification system for meat components in beef and its products

根据可能用于牛肉及其制品掺假的肉类原料(猪肉、鸡肉和鸭肉),设计了4对引物和探针.经过测试其具有良好的特异性后,对影响各自聚合酶链式反应(polymerase chain reaction,PCR)扩增的5个影响因素包括Mg2+浓度、Taq DNA聚合酶活、dNTPs浓度、引物和探针浓度以及模板DNA用量等进行了优化,确定了最佳的PCR扩增体系.同时,根据优化后的结果,对特异性进行进一步的测试,并使用建立的优化荧光PCR鉴别体系对混合样品和市售样品进行检测,确定整体检出限为0.1%(质量分数),并证明该鉴别体系的实际应用价值.

According to the possible meat origin （ pork, chicken, and duck） in beef and their pro-cessed products , specific primers and probes were designed .Following the high specificity confirma-tion experiments, five key factors affecting PCR including TaqDNA polymerase and template DNA a-mounts as well as final magnesiumion （ Mg2＋） , dNTPs and primers concentrations were optimized to establish an optimal real -time PCR system for beef source components identification .Then the speci-ficity was performed againafter optimization .At the same time , mixed samples and commercial beef products were detected by the real -time PCR identification system .The detection limit was 0 .1%and the accuracy was also verified .