Development of a rapid diagnostic test based on loop-mediated isothermal amplification to identify the most frequent...

Identification of Salmonella serovars is performed by conventional seroagglutination or sequencing. These methods are labor-intensive and require technical experience. An easy-to-perform assay allowing the timely identification of the most common non-typhoidal serovars (NTS) is needed. In this study, a molecular assay based on loop-mediated isothermal amplification (LAMP) targeting specific gene sequences of Salmonella Enteritidis, S. Typhimurium, S. Infantis, S. Derby, and S. Choleraesuis has been developed for rapid serovar identification from cultured colonies. A total of 318 Salmonella strains and 25 isolates of other Enterobacterales species that served as negative controls were analyzed. All S. Enteritidis (n = 40), S. Infantis (n = 27), and S. Choleraesuis (n = 11) strains were correctly identified. Seven out of 104 S. Typhimurium and 10 out of 38 S. Derby strains missed a positive signal. Cross-reactions of the gene targets were only rarely observed and restricted to the S. Typhimurium primer set (5 false-positives). Sensitivity and specificity of the assay compared to seroagglutination were as follows: 100% and 100% for S. Enteritidis, 93.3% and 97.7% for S. Typhimurium, 100% and 100% for S. Infantis, 73.7% and 100% for S. Derby, and 100% and 100% for S. Choleraesuis, respectively. With results available in just a few minutes of hands-on time and a test run time of 20 min, the LAMP assay developed here may be a useful tool for the rapid identification of common Salmonella NTS in daily routine diagnostics.