Dual real-time PCR method for rapid detection of sole fish and Pangasius bocourti-derived components in fish products

Abstract

ObjectiveTo establish a dual real-time PCR rapid detection method for sole fish and Pangasius bocourti-derived components.MethodsUniversal 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.ResultsThis 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%.ConclusionThis 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.

Keywords

• sole fish
• pangasius bocourti
• cynogloddus
• real-time pcr
• fish products
• adulteration
• detection