阿胶的半巢式-多重PCR鉴别方法研究

摘要

采用半巢式-多重PCR（聚合酶链式反应）法鉴别阿胶中是否掺杂有马、猪和牛源性成分。首先采用液氮研磨法从阿胶中提取高质量的DNA（脱氧核糖核酸），以通用引物P1和P2进行第一轮扩增，并将扩增产物作为第二轮PCR的模板；再以包含通用引物P1和4种物种（驴、马、猪和牛）特异性引物A、B、C和D的混合引物进行第二轮多重PCR扩增；最后电泳检测第二轮扩增产物，根据电泳条带的分子量大小，判断阿胶产品的动物原材料是否掺假。结果表明，采用本方法可观察到不同动物DNA分子量之间的明显差异，通过DNA电泳结果直接判断阿胶产品中是否掺假。%In this study,semi-nested multiplex PCR method were used to identify whether gelatin doped with horses,pigs and cattle-derived ingredients. Firstly, liquid nitrogen grinding method was used to extract high quality DNA from gelatin. Universal primer P1 and P2 were used for the first round of amplification,and the am-plified product was used as the templates of the second round PCR. Then mixed primers containing universal primer P1 and specific primers (A,B,C and D) was used to do multiplex PCR amplification of second rounds. Final the second round of amplification products with electrophoresis was detected,and authenticity of gelatin was determined according to molecular weight of the electrophoretic bands. The results showed that it can ob-serve the significant differences of the DNA molecular weight of different animals with the method and directly judge whether gelatin is adulterated by DNA electrophoresis results.