Optimization of the reactional medium and a food impact study for a colorimetric in situ Salmonella spp. detection method.

Foodborne pathogens are still a major concern for public health authorities. In this paper, we describe the optimization of a previously reported method which combines a highly specific capture of targeted food pathogens with an intracellular staining method. The reaction medium was optimized to simultaneously allow specific enrichment of Salmonella and maximize the staining of the target pathogen. This in situ colorimetric concept was evaluated with a broad range of food samples artificially contaminated with low levels of stressed Salmonella to mimic natural contamination conditions. This direct detection method compared favorably to a commercially available immunoassay system (Vidas® UP Salmonella), for cooked meat, dry milk powder and egg products. Globally 88% agreement was obtained between the two methods with a sensitivity of 80% and a specificity of 100% for the tested method. Main discordances were obtained with food matrices having high levels of competitive Gram negative microflora. These observations show that the design of an adapted culture medium is necessary to enhance the specific in situ capture and revelation system.

Simplified detection of food-borne pathogens: an in situ high affinity capture and staining concept.

Food industries need simple, rapid and cost-effective solutions for pathogen detection in food and environmental samples. In this paper, we describe a simple but novel detection concept combining an affinity capture surface and intracellular metabolic marker to visualize the bacterial presence on the affinity surface. The surface of a Solid Phase Support (SPS) is functionalized with specific phage tail proteins targeted to the bacterial pathogen of interest. The SPS is placed directly into the primary food enrichment bag after stomaching. Following incubation, the captured bacteria are visually detected in situ as a result of the bacterial reduction of the colorless soluble substrate triphenyltetrazolium chloride (TTC) (present in the primary culture medium) to an intracellular red insoluble formazan product. Detection on the SPS is observed as an intense red color after 22 to 40 hours of enrichment. This is not impaired by the presence of food particles and the natural background microflora. The in situ method significantly simplifies pathogen detection by eliminating any post-enrichment intervention that is necessary in the traditional methods of analysis. We have demonstrated the application of this new approach for the detection of Escherichia coli O157: H7, Listeria spp. and Salmonella spp. in artificially contaminated food samples.