A Rapid Method for Bovine-derived Components Based on Recombinase Polymerase Amplification Technology
The diversification of meat adulteration in the market has made it increasingly difficult for regulatory agencies to identi-fy ingredients of animal origin with ease,accuracy and sensitivity.In particular,beef products with high economic value on the market have become the hardest hit areas for counterfeiting.Therefore,it is urgent to establish a fast and efficient molecular de-tection method for bovine components.Based on the requirement,this study designed and screened several groups of bovine spe-cific amplified primers and probe combinations with β-actin,a reference gene expressed constitutively in cattle.Through sensitivity detection and specificity verification,recombinase polymerase amplification(RPA)method for rapid detection of bovine-derived components was established.At the same time,the efficiency of one-step DNA extraction from beef products was improved by adjusting and optimizing the reagent ratio.PRA technology was combined with colloid gold immunotest strip color develop-ment technology to realize the convenience of detection and visualization of results.Application results showed that the proposed method could detect bovine RPA specifically,with a minimum detection sensitivity of 14.8 copies,and could visualize the visual-ized results obtained by colloidal gold immunotest strips,and the accuracy and sensitivity of the results were the same as that of real-time fluorescence RPA.The method has strong specificity and high sensitivity,and the whole process can be completed in 25 min,greatly shortening the detection time.
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