Independent evaluation of a DNA microarray system for Salmonella detection in ground beef

Abstract

A new DNA microarray test kit has been developed to detect foodborne pathogens in various food matrices. This study focuses on evaluating the PathogenDx microarray-based system to detect Salmonella in ground beef and verify critical parameters that could interfere with the method's effectiveness, such as enrichment incubation time, ground beef fat content, inclusivity, exclusivity, and analytical sensitivity. Sample preparation protocols were evaluated at 6, 8, 12, 18, and 24 h enrichment times at varying bacterial levels to identify optimal conditions to detect the invA gene using the PathogenDx microarray. An 8 h enrichment step was selected based on 100% detection when initial inoculum levels were ≥5 CFU/g, and fractional detection was achieved when the concentration was as low as 1 CFU/g. Thus, the detection of Salmonella using the PathogenDx microarray system can be conducted in 12.5 h, including sample preparation, labeling PCR, hybridization, and analysis. Regarding fat content, there was no significant difference in detection rates of PathogenDx protocol among the highest and lowest commercially sold lean-to-fat ratios of ground beef. Inclusivity and exclusivity experiments showed that Salmonella was correctly identified 100% of the time. Using the ground beef matrix, PathogenDx method is comparable to the United States Department of Agriculture's Microbiology Laboratory Guidebook methodology for detection, which correctly identified Salmonella in 100% of the samples. Salmonella was detected between 93.33 and 100% when ground beef was inoculated with 1 and 5 CFU/g, respectively.