Application of Quadruple Multiplex PCR Detection for Beef, Duck, Mutton and Pork in Mixed Meat

Abstract

In this study, we have developed quadruple multiplex PCR assay for meat (beef, duck, mutton and pork) in processed meat products. By mixing four primers in appropriate ratios could be identified by the PCR. A forward primer was designed on a conserved DNA sequence in the mitochondrial ND2 and 16S rDNA gene in sheep and duck genes, and reverse primers on species-specific DNA sequences for each species. PCR primers were designed to give different length fragments from the four meats. The products showed species-specific DNA fragments of 116, 212, 177 and 322 bp from beef, pork, mutton and duck. Optimal PCR conditions were established. The assay sensitivity under these conditions was 0.1ng, and its specificity was 100 %. The results of the study suggest that PCR represents a simple, efficient test method as a practical alternative for the rapid detection and identification of meat.