Abstract：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 ng～10-5 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.
