生化药物中猪、牛与羊性成分PCR检测的阳性对照品的构建

Construction of Positive Controls for the Determination of Compositions from Bovine,Porcine and Ovine in Biochemical Medicines by PCR

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 develop the positive controls for the determination of compositions derived from bovine, porcine and o- vine in biochemical medicines by PCR to identify the different animal original materials. Methods： DNA was extracted from the self- made positive controls of the compositions from bovine, porcine and ovine tissues, and then was amplified with the specific primers. The amplified products were digested by enzyme, and then the sequence was detected. Results： The length of amplified segments and enzyme-digested segments was the same with that of the target sequence. Conclusion： The developed positive controls for the composi- tions derived from bovine, porcine and ovine can be applied in the identification of biochemical medicines. Promising quality control can be achieved in laboratory for the detection of nucleic acid from bovine, porcine and ovine in biochemical medicines.