Dual real-time PCR method for rapid detection of sole fish and Pangasius bocourti-derived components in fish products
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1.Linyi Inspection and Testing Center, Shandong Linyi 276000, China;2.Qinhuangdao Customs Technology Center, Hebei Qinhuangdao 066004, China;3.Shijiazhuang Customs Technology Center, Hebei Shijiazhuang 050000, China

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R155

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    Abstract:

    Objective To establish a dual real-time PCR rapid detection method for sole fish and Pangasius bocourti-derived components.Methods Universal TaqMan primers and probe sets were designed according to the mitochondrial 16S rRNA gene sequence of 13 species of sole fish. TaqMan primers and probe sets were designed according to the mitochondrial cytb gene sequence of Pangasius bocourti, and the amplification reaction system was optimized for real-time PCR to achieve the purpose of rapid detection of products.Results This method had good specificity, and the sensitivity could reach 10-3 ng sole fish DNA. Sole fish 16S rRNA gene could be detected in fish products mixed with carp meal, and the mass fraction sensitivity could reach 1%. The sensitivity could reach 10-4 ng Pangasius bocourti DNA. Pangasius bocourti cytb gene could be detected in fish products mixed with sole fish, atlantic cod meal and rice noodles, the mass fraction sensitivity of sole fish and atlantic cod meal could reach 0.1%, and the mass fraction sensitivity of rice noodles could reach 0.001%.Conclusion This method had high specificity, high speed and high sensitivity, and could meet the detection requirements of sole fish authenticity and Pangasius bocourti adulteration in fish meat products.

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LI Jie, QIAN Yunkai, WANG Jianchang. Dual real-time PCR method for rapid detection of sole fish and Pangasius bocourti-derived components in fish products[J].中国食品卫生杂志,2022,34(3):539-545.

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History
  • Received:November 16,2021
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  • Online: July 07,2022
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