Proofman Ladder-Shape Melting Temperature Isothermal Amplification for Rapid Detection of Duck-Derived Contaminants in Edible Oils
Abstract
The behavior of mixing fake edible oil is not uncommon, and the addition of "gutter oil" to edible oil will have a serious impact on people's daily lives. A method for detecting duck components in edible oil using Proofman ladder-shape melting temperature isothermal amplification (Proofman-LMTIA) was established by combining Proofman enzyme-mediated probe cleaning with LMTIA technology. Proofman probes and LMTIA primers were designed for specific gene fragments in duck DNA, and the reaction system was optimized through experiments. The results showed that the established Proofman-LMTIA detection method had the best amplification effect at 66 oC and could detect duck DNA concentrations as low as 1 pg/μL. The method has strong specificity and can accurately distinguish duck source genes from DNA of other species, detecting adulteration of 0.1% of duck mass fraction. The amplification time is within 20 min. The established Proofman-LMTIA detection method can quickly detect the adulteration of duck components in edible oil, providing a reference for regulatory authorities.
