Rapid visual detection of beef products adulteration using recombinase polymerase amplification combined with lateral flow dipstick

Beef is one of the most widely consumed meats worldwide, and its products are often adulterated with other meats. To protect consumers' rights and facilitate regulation by relevant government departments, it is important to develop a rapid and accurate method to detect adulteration in meat and meat products. In this study, we employed bioinformatics methods to identify specific sequences of cattle, pig, chicken, and duck, and designed primers and probes accordingly. A method based on recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD) was developed for rapid visual detection of the authenticity of beef and beef products. The RPA reaction was conducted at 37 degrees C for 20 min. The amplification products were then diluted and applied to the sample pad of the LFD. Results were visible to the naked eye within 5 min. The results demonstrated that the method could specifically differentiate components of bovine, porcine, chicken, and duck origin, with a limit of detection (LOD) of approximately 20 copies for each species. The method was applied to 10 commercially available beef products. Of which, five samples were detected with porcine-derived components. The results of the RPA-LFD method were verified using PCR and observed to be consistent between the methods. Compared with other methods, this method is easy to use, requires no specialized equipment, and delivers results in about 30 min from amplification to detection, making it suitable for rapid visual detection on-site. Therefore, it can serve as a technical reference for detecting adulteration in meat and meat products.