Establishment of ERA Visual Rapid Detection Methods for Salmonella

In order to realize the rapid detection of Salmonella in food, in this study, the Salmonella RPA primers and probes reported in the literature were used to establish the enzymatic recombinase amplification (ERA) fluorescence method and chromogenic method for visualization of Salmonella. Then, the specificity, sensitivity and detection limit of the two methods were analyzed. Furthermore, considering the requirements of the on-site detection environment outside the laboratory, the self-heating box and heat stickers were used as heat sources for DNA extraction and ERA reaction on-site, respectively. The optimized detection time was only 11 min and 4 min respectively, and the sensitivity was up to 10- 2 ng/µL. The detection limit of artificially contaminated samples was 1 CFU/mL after culture for 4 h. This study innovatively applied RPA primers and probe applied to ERA detection system, established two visual rapid detection methods: fluorescence method and chromogenic method, and integrated on-site DNA extraction and ERA reaction devices. In this study, the Salmonella RPA primer probe was innovatively applied to the ERA detection system, two visual and rapid detection methods, fluorescence method and chromogenic method were established, and the on-site DNA extraction and ERA reaction detection kit were integrated to improve the amplification efficiency. And get rid of the dependence on traditional laboratory equipment, it is of great significance to promote the rapid detection of food-borne pathogens on-site visualization. © 2023 Chinese Institute of Food Science and Technology. All rights reserved.