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 (m-PCR) 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.