牛肉及其制品中肉类掺假的荧光PCR鉴别体系优化

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.

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

According to the possible meat origin (pork, chicken, and duck) in beef and their processed products, specific primers and probes were designed. Following the high specificity confirmation experiments, five key factors affecting PCR including TaqDNA polymerase and template DNA amounts 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 specificity 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.