Detection of Soybean-Derived Components in Dairy Products Using Proofreading Enzyme-Mediated Probe Cleavage Coupled with Ladder-Shape Melting Temperature Isothermal Amplification (Proofman–LMTIA)
Fugang Xiao,
Menglin Gu,
Yaoxuan Zhang,
Yaodong Xian,
Yaotian Zheng,
Yongqing Zhang,
Juntao Sun,
Changhe Ding,
Guozhi Zhang,
Deguo Wang
Affiliations
Fugang Xiao
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Menglin Gu
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Yaoxuan Zhang
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Yaodong Xian
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Yaotian Zheng
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Yongqing Zhang
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Juntao Sun
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Changhe Ding
College of Grain and Food, Henan University of Technology, Zhengzhou 450001, China
Guozhi Zhang
College of Grain and Food, Henan University of Technology, Zhengzhou 450001, China
Deguo Wang
Henan Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety, Food and Pharmacy College, Xuchang University, Xuchang 461000, China
Food adulteration is a serious problem all over the world. Establishing an accurate, sensitive and fast detection method is an important part of identifying food adulteration. Herein, a sequence-specific ladder-shape melting temperature isothermal amplification (LMTIA) assay was reported to detect soybean-derived components using proofreading enzyme-mediated probe cleavage (named Proofman), which could realize real-time and visual detection without uncapping. The results showed that, under the optimal temperature of 57 °C, the established Proofman–LMTIA method for the detection of soybean-derived components in dairy products was sensitive to 1 pg/μL, with strong specificity, and could distinguish soybean genes from those of beef, mutton, sunflower, corn, walnut, etc. The established Proofman–LMTIA detection method was applied to the detection of actual samples of cow milk and goat milk. The results showed that the method was accurate, stable and reliable, and the detection results were not affected by a complex matrix without false positives or false negatives. It was proved that the method could be used for the detection and identification of soybean-derived components in actual dairy products samples.
