Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (9): 1873-1880.doi: 10.3864/j.issn.0578-1752.2012.09.024

• RESEARCH NOTES • Previous Articles     Next Articles

A Quick Multiplex PCR Method for the Identification of Four Meat Ingredients in Food Products

 HE  Wei-Ling, ZHANG  Chi, YANG  Jing, HUANG  Ming, YANG  Jun   

  1. 1.南京农业大学食品科技学院/肉品加工与质量控制教育部重点实验室，南京 210095
    2.南京市产品质量监督检验院，南京  210028
  • Received:2011-06-16 Online:2012-05-01 Published:2011-09-28

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Abstract: 【Objective】An accurate and reliable universal primers-multiplex PCR (UP-M-PCR) method was developed to identify 4 kinds of meat ingredient (chicken, beef, lamb, pork) in food products.【Method】 Total DNA in meat samples were extracted using a commercial DNA extraction kit, a CTAB-Proteinase k method and a SDS-Proteinase k method, respectively. The concentration and purity of DNA were evaluated to select the optimal method. Two sets of UP-M-PCR primers in different lengths were designed based on the homologous and specific sites of mitochondrial cytochrome b gene. Each set was comprised of 5 primers: one general forward primer and four species-specific reverse primers. The UP-M-PCR system, followed by gel electrophoresis assay of speces-specific bands, was established for meat identification. The meat spices were identified according to different lengths of PCR products. Meanwhile, the accuracy of the method was evaluated by testing of eighty food samples from the market. 【Result】The SDS-Proteinase K method and the DNA commercial kit presented better extraction efficiencies in contrast to the CTAB-Proteinase K method. The optimized UP-M-PCR was established using the set of longer primes, which showed higher specificity than that using the shorter ones. The detection limit was picograms of DNA. The accuracy of the method was verified by examination of 80 food samples from the market. 【Conclusion】A sensitive, reliable and convenient method was developed to identify 4 kinds of meat ingredient in food products.

Key words: multiplex PCR, identification, pork, beef, lamb, chicken

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