Establishing a Detection Method Based on Multiplex PCR for Identification of Sheep Meat, Goat Meat and Common Adulterant Meats

This study aimed to establish a multiplex PCR identification system capable of rapidly detecting adulteration in sheep and goat meat, while qualitatively identifying common adulterant meats (pork, chicken, and duck). Species-specific primers targeting mitochondrial DNA sequences were designed after screening for gene fragments with intraspecies conservation and interspecies specificity across five target species. The multiplex PCR conditions and system were systematically optimized and evaluated for specificity, reproducibility, sensitivity, and practical applicability using simulated mixed samples and heat-treated products. The results demonstrated that the system could successfully identify sheep meat, goat meat, and adulterant meat components in randomly combined target meat template DNAs with excellent reproducibility. The system maintained a high sensitivity, detecting target species even at low DNA template concentrations and in samples with low adulteration ratios. Moreover, target meat components remained detectable in heat-treated products, confirming the system's robustness under realistic market conditions. This multiplex PCR identification system demonstrates strong specificity, good reproducibility, high sensitivity, and broad applicability. It provides an important tool for effectively monitoring sheep and goat meat adulteration and offers crucial technical support for ensuring the authenticity of sheep and goat meat.