Rapid and simple detection of the invA gene in Salmonella spp. by isothermal target and probe amplification (iTPA)

Abstract

Salmonella spp. are an important cause of food-borne infections throughout world, and the availability of rapid and simple detection techniques is critical for the food industry. Salmonella enterica serovars Enteritidis and Typhimurium cause the majority of human gastroenteritis infections, and there are a reported 40,000 cases of salmonellosis in the United States each year. A novel rapid and simple isothermal target and probe amplification (iTPA) assay that rapidly amplifies target DNA (Salmonella invA gene) using a FRET-based signal probe in an isothermal environment was developed for detection Salmonella spp. in pre-enriched food samples. The assay was able to specifically detect all of 10 Salmonella spp. strains without detecting 40 non-Salmonella strains. The detection limit was 4 x 10¹ CFU per assay. The iTPA assay detected at an initial inoculum level of <10 CFU in the pre-enriched food samples (egg yolk, chicken breast and peanut butter). This detection system requires only a water bath and a fluorometer and has great potential for use as a hand-held device or point-of-care-testing diagnostics. The iTPA assay is sensitive and specific and has potential for rapid screening of Salmonella spp. by food industry.