A novel common single primer multiplex polymerase chain reaction (CSP-M-PCR) method for the identification of animal species in minced meat

摘要

BACKGROUND: Minced meat species adulteration is a severe problem. In this study a common single primer multiplex polymerase chain reaction (CSP-M-PCR) method was applied to the identification of five species (chicken, cattle, sheep, pig and horse) in minced meat products. CSP was designed as a common adapter at the 5'-end of species-specific reverse primers and employed in giving different length fragments from the respective meat species.RESULTS: Species-specific DNA fragments could be identified in a single PCR reaction by mixing CSP, Chicken-R, Cattle-R, Sheep-R, Pig-R and Horse-R primers in the ratio 1:0.01:0.01:0.01:0.01:0.10 (where '1' represents 0.5 mmol Lp#). The specific DNA fragments could be efficiently amplified by CSP-M-PCR with > 5 omol Lp# species-specific primers, except for 50 omol Lp# Horse-R. A detection limit of 0.5 g kgp# was determined for both single species of minced meat and all species of five kinds of minced meat.CONCLUSION: The CSP-M-PCR method simplified the PCR reaction system and removed the inconsistent amplification efficiency from different primers. This highly sensitive, reproducible and rapid method could potentially be used as a screening or identifying assay to test for the presence of species or ingredients in minced meat and other meat products.