Recombinase Polymerase Amplification Based Multiplex Lateral Flow Dipstick for Fast Identification of Duck Ingredient in Adulterated Beef

SIMPLE SUMMARY: The adulteration and authenticity of meat and meat products has become a global social problem. Beef is often intentionally adulterated with cheap meat. In order to ensure the authenticity of meat, and provide technical support to regulatory authorities, we developed a rapid and visual method to detect duck ingredient in adulterated beef. This method is implemented recombinase polymerase amplification (RPA) and multiplex lateral flow dipstick (MLFD) cascade. The whole RPA-MLFD reaction process can be finished within 35 min, and the results can be determined by naked eyes. RPA-MLFD was applied to simultaneously detect duck ingredient and beef ingredient without using additional instruments. An adulteration ratio as low as 5% of duck ingredient in beef can be easily measured. Moreover, we confirmed that our new method held good potential in the detection of commercially processed meat samples. Therefore, this study reports a useful animal derived meat adulteration detection method, which have potential application in future. ABSTRACT: Meat adulteration has become a global social problem. In order to protect consumers from meat adulteration, several methods have been developed to identify meat species. However, the conventional methods are labor-intensive, time-consuming and require instruments. In the present study, a rapid and visual method based on recombinase polymerase amplification (RPA) and multiplex lateral flow dipstick (MLFD) was developed to detect duck ingredient in adulterated beef. Using recombinase and strand displacement polymerase enable RPA to amplify different double-labeled DNA amplicons at room temperature, which can be further detected by MLFD. The whole reaction process can be finished within 35 min, and the results can be determined by naked eyes. As low as 5% of duck ingredient in adulterated beef can be easily measured. Moreover, we confirmed that our new method held good potential in the detection of commercially processed meat samples. In conclusion, this study reported a useful animal derived meat adulteration detection method, which have potential application in future.