Detection of Pork-Derived Ingredients Based on Real-time Quantitative PCR

In order to accurately and reliably quantify pork-derived ingredients in meat products, single-copy gene (carcinoembryonic antigen-related cell adhesion molecule 2-like, CACA) in pig cell nucleus was screened by bioinformatics methods, with CACA gene as the amplification target, specific primers and TaqMan probes were designed, and a TaqMan real-time quantitative PCR method for detecting pork ingredients was established. The experimental results showed that the method had good extraspecies specificity and intraspecies stability. There was a good linear relationship between the template DNA and the amplified Ct value within the DNA concentration range from 20 to 0.032 ng/µL. The standard curve was y=−3.508x+28.636, the coefficient of determination (R²) value was 0.997. Under the condition of relative standard deviation ≤25%, it could accurately detect DNA samples with pork DNA content as low as 0.1%, and meat products with pork content as low as 5.0%. Through testing of commercially available samples, it was found that somemeat products were mixed with pork but were not labeled. And some meat products, pork was not listed as the main ingredient but in fact pork was the main ingredient. The real-time quantitative PCR method established in this study could be used to accurately and reliably detect pork ingredients in meat products. It has important reference value for accurately identifying adulteration of meat products and quantifying the ingredients of meat products.