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目的:建立生化药物中的猪、牛与羊性成分PCR检测方法的阳性对照品,用于检测生产原料的动物来源是猪源性、牛源性或羊源性。方法:分别对自行制备的猪、牛及羊源性成分的阳性对照品,进行了DNA提取,并加入猪、牛与羊特异性引物进行扩增。对扩增产物进行酶切及测序验证。结果:猪、牛及羊阳性对照扩增产物片段长度、酶切片段长度均与目标序列长度一致。结论:所构建的猪、牛与羊性成分的阳性对照品可广泛应用于生化药物的种属鉴别中,对生物药物中猪、牛与羊性成分的核酸检测可进行良好的实验室质量控制。
OBJECTIVE: To establish a positive control for PCR detection of sex components of swine, cattle and sheep in biochemical medicines. The animal origin used for the detection of raw materials for production is pig, bovine or sheep origin. Methods: DNA samples were extracted from positive control samples of pigs, cattle and sheep, respectively, which were prepared by themselves. The specific primers of pig, cattle and sheep were added for amplification. The amplified product was digested and sequenced. Results: The length of amplified products and the length of digested fragments of positive control pigs, cattle and sheep were all consistent with the target sequences. Conclusion: The positive control of pig, cow and sheep can be widely used in the identification of biochemical drugs in the species, nucleic acid detection of the sex components of swine, cattle and sheep in biological drugs can be a good laboratory quality control .