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Species Identification of Fox-, Mink-, Dog-, and Rabbit-Derived Ingredients by Multiplex PCR and Real-Time PCR Assay

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Abstract

Various detection methods have been developed to date for identification of animal species. New techniques based on PCR approach have raised the hope of developing better identification methods, which can overcome the limitations of the existing methods. PCR-based methods used the mitochondrial DNA (mtDNA) as well as nuclear DNA sequences. In this study, by targeting nuclear DNA, multiplex PCR and real-time PCR methods were developed to assist with qualitative and quantitative analysis. The multiplex PCR was found to simultaneously and effectively distinguish four species (fox, dog, mink, and rabbit) ingredients by the different sizes of electrophoretic bands: 480, 317, 220, and 209 bp. Real-time fluorescent PCR’s amplification profiles and standard curves showed good quantitative measurement responses and linearity, as indicated by good repeatability and coefficient of determination R 2 > 0.99. The quantitative results of quaternary DNA mixtures including mink, fox, dog, and rabbit DNA are in line with our expectations: R.D. (relative deviation) varied between 1.98 and 12.23% and R.S.D. (relative standard deviation) varied between 3.06 and 11.51%, both of which are well within the acceptance criterion of ≤ 25%. Combining the two methods is suitable for the rapid identification and accurate quantification of fox-, dog-, mink-, and rabbit-derived ingredients in the animal products.

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Acknowledgements

This study was supported by Major International Cooperation NSFC (31210103917).

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Authors and Affiliations

  1. Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, 430070, China

    Qingqing Wu, Shengnan Xiang, Wenjun Wang, Yueran Zhen & Bang Liu

  2. Department of Animal Technology, Henan University of Animal Husbandry and Economy, Zhengzhou, Henan Province, China

    Jinyan Zhao

  3. Department of Biology Science, Shangrao Normal College, Shangrao, Jiangxi Province, China

    Jinhua Xia

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  1. Qingqing Wu
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  2. Shengnan Xiang
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  3. Wenjun Wang
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  4. Jinyan Zhao
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  5. Jinhua Xia
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  6. Yueran Zhen
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  7. Bang Liu
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Correspondence to Bang Liu.

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Wu, Q., Xiang, S., Wang, W. et al. Species Identification of Fox-, Mink-, Dog-, and Rabbit-Derived Ingredients by Multiplex PCR and Real-Time PCR Assay. Appl Biochem Biotechnol 185, 1–12 (2018). https://doi.org/10.1007/s12010-017-2621-2

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  • DOI: https://doi.org/10.1007/s12010-017-2621-2

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