Preliminary establishment of a multiplex PCR method for the identification of pork and beef meat based on cytochrome-B Gene

Abstract

Molecular species detection in food has become common in the last years. In this study, multiplex PCR (mPCR) technique was applied to discriminate between the pork and beef with the aim of detecting the pork in food products made from beef. We developed a m-PCR protocol detected the presence of pork in food products made from beef based on Cytochrome-b gene by a general primer pair F and primer pair (RP, RC), specific two private with pig and cow. The m-PCR method was successfully designed with technical parameters, such as the annealing temperature of 590C and the final concentration of each primer in a reaction of 0,4µM. The minimum DNA concentration of pig and cow could be detected by m-PCR, which was 0,1ng/µl. This process was tested on 24 different beef sausage samples tagged no pork, resulting in 11/24 (46%) of samples, were found the presence of pork and 54% (13/24) of samples no beef and pork. According to this sequencing result that are completely accordant, we affirm primer-specific amplification in this study can be applied to experiment on large number of sample and the ton other types food made from beef.

Keywords

• beef
• cytochrome-b
• discriminate
• multiplex pcr
• pork