Abstract
Adulterating undeclared animal components into meats may cause health problems. In this study, nuclear DNA sequences for specific detection of sheep/goat, fox and murine (mouse, rat and hamster) were selected to design primers and probes for determination of mutton adulteration. The inter-specific specificity and intra-specific conservation were strictly evaluated in different individuals of 19 animal species. These three nuclear DNA sequences were confirmed to be fixed copy sequences by relative quantitative PCR. Furthermore, a multiplex qualitative and quantitative PCR system with sensitivity of 0.05 ng was developed to efficiently discriminate sheep/goat (237 bp), fox (211 bp) and murine (160 bp) in one reaction. Finally, several mixed DNA samples of goat, fox and rat were quantitatively analyzed by multiplex real-time PCR method, showing good accuracy (R.E. from 1.64% to 18.43%) and precision (R.S.D. from 0.51% to 12.70%). The method developed in this study will provide a powerful technology to combat meat adulteration and protect consumer health.
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