A highly specific PCR assay for identification of raw and heat treated mutton (Ovis aries)

A highly species-specific polymerase chain reaction (PCR) was developed for authentic identification of raw and heat treated sheep meat (mutton). Oligonucleotide primers were designed against a unique target region of sheep mitochondrial D-loop, that yielded four PCR products upon gel electrophoresis of which two amplicons of molecular size 329 and 404 bp were found highly diagnostic. Chances of cross-amplification were precluded by considering as many as 25 species and suitability of the developed sheep-specific PCR was confirmed in raw (n = 20), cooked (60, 80 and 100 °C for 30 min), autoclaved (121 °C for 30 min) and microwave processed meats. Further, the sensitivity of the assay was established to be 0.1% for the detection of adulteration, while limit of detection of ovine DNA was as low as 1 pg. This PCR technique has applications in solving problems related to meat adulteration, quality assurance, forensics, mutton authentication and traceability.