Multiplex PCR assay for species identification of meat and dairy products from buffalo (Bubalus bubalis), cattle (Bos indicus and Bos taurus), goat (Capra hircus), and sheep (Ovis aries)

Abstract

Cases of fraudulent meat and dairy products have increased worldwide, especially in developing countries. To determine the misrepresented animal species, appropriate tools in routine monitoring should be available for food inspections. In the present work, a multiplex polymerase chain reaction assay for species identification of products from ruminants including buffalo, cattle, goat, and sheep was developed. The primer set KUMUT_cFarmSp1 was composed of five species-specific primers and a pair of positive-control primers. The primer set amplified 106-, 163-, 232-, and 308-bp specific fragments from the cytochrome b (cyt b) gene of buffalo, cattle, goat, and sheep, respectively, and 370-bp positive-control fragment from 16S ribosomal RNA (16S rRNA). The detection limit of this PCR assay is 0.1 ng of DNA template. The developed primer set exhibited strong specificity, sensitivity, robustness, and simplicity for food verification, thus indicating its usefulness for species verification in food quality control and law enforcement.