Abstract: 【Objective】To improve the existing method and develop a fast, precise and reliable PCR method for detection of animal derived materials in feedstuff. 【Method】Firstly, general primer designed according to 16S rRNA gene sequence of bovis, sheep, pig, chicken, fish and horse mtDNA was used for primary detection of animal derived materials in feedstuff. Species-specific primers designed according to conserved sequence of mtDNA of bovis, sheep, pig and chicken were used for amplification of a 271bp, 274bp, 149bp and 266bp fragment respectively. Further confirmation of the detection result was then performed. 【Result】PCR method for detection of animal derived materials in unknown feedstuff was developed by using general primer, relevant PCR system and PCR condition. Also a PCR method for detection of each species (bovis, sheep, pig and chicken) was designed by using our species-specific primers. High sensitivity and specificity of our method was confirmed with a minimum detection level of 0.1%. 【Conclusion】Method for detection of animal derived materials in this research is not only cheap and easy for operation, but also precise and reliable for result. It could be one of the effective methods for detection animal derived materials in feedstuff.

Key words: Animal feedstuff, Sensitivity and specificity detection, PCR, Primer