Abstract
It is possible and risky for fur animal carcasses to be mixed into meat products, which is a potential danger for meat quality safety and human health. Therefore, meat validation of quality and constituents is crucial. A variety of methods have been developed to identify muscle tissues of different species. However, most of these methods have the disadvantages of poor repeatability, complex operation and low efficiency, and cannot simultaneously detect multiple species of muscle tissue. The purpose of this study was to construct a multiplex PCR protocol to detect the samples of mink, fox, and raccoon dog. In this study, the specific primers of mink, fox, and raccoon dog were designed according to the variable region sequence of mitochondrial cytochrome b (Cytb) gene. The primers showed good specificity and 50 ℃ was determined as the optimal annealing temperature. The lowest concentration of DNA template of mink, fox, or raccoon dog that could be determined simultaneously by a single tube was 1 pg/µL. Clinical tissue samples detect analysis test results showed that this method could identify whether the tissue samples of three fur animals were mixed from the muscles of chickens, ducks, dogs, cattle, sheep, pigs and rabbits in one PCR reaction simultaneously. In conclusion, the scheme exhibited the advantages of convenient operation, low cost, strong species specificity, high sensitivity, good stability, and repeatability. The systematic optimized inspection process can be applied to meat detection to ensure veterinary public health safety, which has important scientific significance, production, public health, and safety significance.
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All data generated or analyzed during this study are included in this published article and its supplementary information files. The datasets are also available from the corresponding author on reasonable request.
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Funding
This work was mainly supported by Major Science and Technology Innovation Project in Shandong Province (2023CXGC010704), Shandong Province Modern Agricultural Technology System Innovation Team Program (SDAIT-21–10), the Earmarked Fund for the Protection for the State Forest and Wild Animals (No. 2130211), and Funds of Shandong “Double Tops” program.
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Huijun Guo conceived and designed the experiments. Yao Pan, Xuemin Wang and Wenjie Ma are mainly responsible for experimental implementation. Hongmei Li, Jianhua Qiu and Yuxin Sun is mainly responsible for sample collection and helped to do some experiments. Jianhua Qiu, Xuemin Wang and Yao Pan wrote the manuscript. All authors have read and approved the final manuscript
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Yao Pan declares that he has no conflict of interest. Xuemin Wang declares that she has no conflict of interest. Wenjie Ma declares that he has no conflict of interest. Hongmei Li declares that she has no conflict of interest. Yuxin Sun declares that he has no conflict of interest. Jianhua Qiu declares that he has no conflict of interest. Huijun Guo declares that he has no conflict of interest.
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Ethical approval was not required for the study because the sampling process did not harm the animals.
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Pan, Y., Wang, X., Ma, W. et al. Establishment and Application of Multiplex PCR Scheme for Simultaneously Distinguishing Muscle Tissues of Mink, Fox, and Raccoon Dog from Conventional Livestock and Poultry. Food Anal. Methods 17, 226–235 (2024). https://doi.org/10.1007/s12161-023-02560-y
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DOI: https://doi.org/10.1007/s12161-023-02560-y