Specific Identification of the Adulterated Components in Beef or Mutton Meats Using Multiplex PCR

Background: The fraudulent substitution of cheap and low-quality meat for expensive and good quality meats to gain profit is a common practice in industries worldwide. Adulteration of fox, raccoon, or mink in commercial beef and mutton meat in the supermarket has become a serious problem. Objective: To ensure the meat quality and safety, we have developed a multiplex PCR method to identify the fox, mink, and raccoon components adulterated in beef or mutton with very low detection limits. Methods: PCR primers were designed and tested by examining the size of PCR product, the nuclease digestion products, and DNA sequencing. Results: After primer interference tests, we have established a double PCR method that can clearly identify fox, mink, or raccoon components in beef meat and mutton meat at the 1% (w/w) level. Triplex PCR and quadruple PCR have been also developed, which are able to identify any two types of components or three mixed components in beef meat unambiguously. Conclusions: We have developed multiplex PCR systems. The duplex PCR systems can identify one component (fox, raccoon, or mink) adulterated in beef meat or mutton meat without question, and triplex PCR and quadruple PCR can discriminate two components and three components adulterated in beef meat. Highlights: These methods are convenient, low-cost, highly specific and reliable, and of a great value for meat quality control and food safety quarantine.