Duplex Real-time PCR Methods for Quantitative Detection of Bovine Derived Materials in Animal Products

doi:10.13560/j.cnki.biotech.bull.1985.2017-1129

Abstract

Abstract: The objective of this work is to develop a duplex real-time quantitative PCR method for detecting the amount of bovine derived material in animal product. Using bovine satellite IV DNA as detection target gene and myostatin gene as an internal control to synthesize primer probe,the standard freeze-dried meat mixture powder with known beef proportion was detected by duplex real-time PCR based on TaqMan,and the linear model was established. Then the model was verified by detecting mocked and delivered samples. The method detected 0.1% bovine material in freeze-dried meat powder,and no cross reaction with the tissues of other animal species occurred. The relative standard deviation of repeatability test was < 5%,and the amplification efficiency of the method was 93.6%. Conclusively,the established method is suitable for quantitative detection of bovine derived material in meat products,and can be used to determine the content of bovine derived material in animal products on the market.

Key words: duplex real-time PCR, quantitative detection, bovine derived material

GAO Zhi-qiang, WANG Lin, PU Jing, YIN Yi, ZHANG Wei, ZHAO Xiang-peng, YAO Zhen-yu. Duplex Real-time PCR Methods for Quantitative Detection of Bovine Derived Materials in Animal Products[J]. Biotechnology Bulletin, 2018, 34(9): 190-194.

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