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| Development of Proofman-Ladder-Shape Melting Temperature Isothermal Amplification Method for Detection of Pig-Derived Components in Meat Products |
| WANG Borui, XU Dandan, GU Menglin, YAO Wei, WANG Yao, XIAO Fugang, WANG Deguo |
| 1.College of Food and Bioengineering, Henan University of Science and Technology, Luoyang 471000, China; 2.Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety of Henan Province, College of Food and Pharmacy, Xuchang University, Xuchang 461000, China; 3.College of Food Science and Engineering, Henan University of Technology, Zhengzhou 450001, China |
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Abstract The study aimed to establish a proofreading enzyme-mediated probe cleavage combined with ladder-shape melting temperature isothermal amplification (Proofman-LMTIA) method for detection of pig-derived components in meat products. The LMTIA primers and Proofman probes were designed using the porcine prolactin receptor (PRLR) gene as the target. The LMTIA reaction system was optimized, and the specificity, sensitivity and limit of detection (LOD) of the proposed method were determined. The results showed that the amplification could be completed within 30 min, and the Proofman-LMTIA method could specifically detect the genomic DNA of pigs rather than of chicken, ducks, cattle, sheep, dogs, sweet potato, cassava, mung bean and wheat. The sensitivity was 1 ng/μL, and the LOD for artificially adulterated meat samples was 0.1%. The Proofman-LMTIA method has high specificity for pig-derived components, and can quickly and accurately detect pig-derived components in meat products and rapidly detect meat adulteration.
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2023, Vol. 37 
