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

Abstract

Abstract 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 L −1 ). The specific DNA fragments could be efficiently amplified by CSP‐M‐PCR with > 5 µmol L −1 species‐specific primers, except for 50 µmol L −1 Horse‐R. A detection limit of 0.5 g kg −1 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. Copyright © 2008 Society of Chemical Industry