Simultaneous Identification of Water Buffalo, Cattle, and Yak Meat and Horns by Multiplex Polymerase Chain Reaction (PCR)

Beef is an important source of protein for human consumption globally; in addition to beef, water buffalo and yak meat are widely consumed in Asia. Additionally, water buffalo horns are an integral part of traditional Chinese medicine. Adulteration of water buffalo horn with cattle and yak horn for economic gain has become a prevalent and pressing concern. This study used multiplex polymerase chain reaction (PCR) for the simultaneous identification of meat and horns from water buffalo, cattle, and yaks within a single PCR test. Species-specific primers were designed for multiplex PCR based on a comprehensive comparison of mitochondrial gene sequences with those of commonly edible and horned animals. Multiplex PCR conditions were optimized by considering the concentration of components and the annealing temperatures for each set of species-specific primer pairs. A total of 220 bp amplicons were obtained for water buffalo, 269 bp for cattle, and 346 bp for yak. Validation measures were used to eliminate interference from other animal sources. Simultaneous identification of the three species in a single assay presents substantial advantages over conventional species-specific PCR methods.