Beef specific polymerase chain reaction assay for authentication of meat and meat products

The aim of this study was to develop species-specific polymerase chain reaction (PCR) assay for specific detection of beef using self-designed primer pair based on D-loop region of mitochondrial gene for amplification of 513 bp DNA fragments from fresh, processed and autoclaved meat and meat products. The beef-specific primer pair was self-designed based on the available gene sequences on NCBI nucleotide database. The primer pair was individually optimized for amplification of desired 513 bp DNA fragments from isolated DNA of fresh beef. After successful amplification of desired DNA fragments by this primer pair, the PCR assay was evaluated for their efficiency to amplify DNA extracted from cooked and autoclaved meat and meat emulsion. The level of detection of this beef-specific primer pair was found to be less than 1 percent using PCR assay, even in admixed meat products containing meat of beef, buffalo meat, pork, chevon, mutton and chicken. No adverse effect of heat treatment, processing conditions and ingredients was observed on amplification pattern. The experiments were repeated for several time and results was found to be repeatable every-time.