Multiplex TaqMan locked nucleic acid real-time PCR for the differential identification of various meat and meat products

Meat adulteration incidents have been reported frequently over the past few years, and the corresponding traceability issues attracted much more attention due to the customer's demands and administration's responsibility. Therefore, it is important to develop high-throughput and rapid detection methods to identify the specific sources from meat samples. In this study, a multiplex TaqMan locked nucleic acid real-time polymerase chain reaction assay (MLNA-RT-PCR) was developed to simultaneously detect multiple meat sources (duck, pork, beef and chicken). PCR primers and TaqMan-LNA probes were designed based on species-specific mitochondrial gene sequences, and the MLNA-RT-PCR was developed and optimized for better performance. The specificity of this assay was verified through identifying unrelated (sheep, horse, deer, donkey, rabbit, goose, goat, shrimp, salmon and maize) mitochondrial DNA as species-specific targets. The detection limit for MLNA-RT-PCR reached to the level of 0.01% of each species. The assay was then used to identify the meat sources of commercial meat and meat-derived products that were obtained from markets in Shantou, and the results were 98% consistent with that obtained from detection based on the national standard. In conclusion, this MLNA-RT-PCR is a high-throughput, sensitive and specific method that can be used to identify multiple meat sources in meat and meat-derived products.