A Multiplex Real-Time PCR for Qualitative Detection of Mutton, Pork, Chicken, and Duck in Processed Meat Products

Economically motivated adulteration (EMA) is a persistent global issue in the meat industry. This study aimed to develop a qualitative detection method for mutton, pork, chicken, and duck meat based on nuclear genes. The specificity of the method was validated by amplifying DNA from 23 samples including animal, plant, and microbial mixtures. Accuracy was confirmed by testing 11 reference samples, which produced the expected results. Sensitivity tests demonstrated a limit of detection as low as 10 copies for mutton, pork, and chicken, and 20 copies for duck. To assess the applicability of this method to real-world samples, 35 processed meat products were tested using the established multiplex real-time polymerase chain reaction (PCR) method. Labeling information for 34 of the 35 products matched the detection results, with one exception: a mutton roll contained no detectable mutton. Therefore, the developed multiplex real-time PCR method demonstrated good specificity, accuracy, and sensitivity, making it a reliable tool for the qualitative detection of mutton, pork, chicken, and duck in processed meat products.