Semi-nested PCR检测饲料中牛羊源成分

加强对进口饲料中牛羊源成分的检测是防止疯牛病和痒病传播的一个重要措施。根据已发表的牛和羊特异性基因及引物序列，分别设计了1条牛和羊特异性semi-nested PCR引物，并采用semi-nested PCR技术对饲料中的牛和羊成分进行了扩增检测。结果表明，semi-nested PCR能够扩增得到247 bp的牛特异性基因条带和214 bp的羊特异性基因条带，其对饲料中牛或羊源性成分的检测灵敏度可达到0.00001%～0.0001%，比普通PCR检测灵敏度要高出103倍；对牛或羊成分DNA的检测灵敏度可以达到10-6～10-5 ng，比普通PCR检测灵敏度要高出105倍以上。该技术具有快速、灵敏和结果稳定的特点，是检测饲料中痕量牛羊源成分的一种有效方法。

Identification of Bovine and Sheep Derived Materials in Feedstuff by Semi-nested PCR

Augment identification of bovine and sheep derived materials in feedstuff is essential for effective control transmission of bovine spongiform encephalopathy and scrapie. This study described a semi-nested PCR assay for the detection of bovine and sheep materials in feedstuff. Bovine- or sheep-specific semi-nested PCR primer had been designed for this purpose based on their published species-specific gene and primer's sequences. The results of semi-nested PCR showed that 247 and 214 bp bands were obtained from the bovine and sheep materials respectively, the detection limitation was 0.00001%0.0001% for bovine and sheep derived materials in feedstuff and 10-6 ng10-5 ng for bovine and sheep genomic DNA respectively, 103 and 105 times higher than that of the detection limitation of PCR. Therefore, the rapid, sensitive and stable semi-nested PCR are the effective methods for detecting trace bovine and sheep derived materials in feedstuff.