Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (21): 4578-4585.doi: 10.3864/j.issn.0578-1752.2013.21.021

• STORAGE·FRESH-KEEPING·PROCESSING • Previous Articles     Next Articles

Establishment of a Taqman Real-time PCR with Internal Amplification Control for the Detection of Chicken and Pork Ingredient in Food Products

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.河南省商丘市技术监督局/国家面粉及制品质量监督检验中心，河南商丘 476000   

  1. 1.College of Food Science and Technology, Nanjing Agricultural University/Key Laboratory of Meat Processing and Quality  Control, Ministry of Education, Nanjing 210095
    2.Shangqiu Bureau of Quality and Technical Supervision/National Flour and Flour Products Supervision and Inspection Center, Shangqiu 476000, He’nan
  • Received:2012-10-25 Online:2013-11-01 Published:2013-07-25

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Abstract: 【Objective】A Taqman real-time PCR with IAC (Internal amplification control) was developed to identify pork and chicken ingredient in meat products. 【Method】 Specific primers and probes were designed based on the sequence of beta actin gene of pig DNA and transforming growth factor gene of chicken DNA, respectively. The specificity and sensitivity of the primers and probes were evaluated. An IAC was designed and constructed, and the real-time PCR system with IAC was established by optimizing the concentration of IAC in the system. The DNA of raw and cooked samples was evaluated by applying the Taqman real-time PCR system with IAC. Meanwhile, 38 food samples from market were assayed. 【Result】The pork and chicken ingredient in food was detected effectively by the Taqman real-time PCR with IAC. No cross-reaction was observed between species specific primer-probe systems in 40 cycles. The limit of detection was 0.5 ng of template DNA. No significant difference was observed between the Ct values of raw and cooked samples. The accuracy of the method was verified by examination of 38 food samples from the market.【Conclusion】An accurate and reliable method was developed to identify pork and chicken ingredient in food products.

Key words: Internal amplification control (IAC) , detection , real-time PCR , chicken ingredient , pork ingredient

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