Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6288650 | Food Microbiology | 2014 | 38 Pages |
Abstract
Multiple drug resistance in Salmonella is an emerging problem in the area of food safety. Depending on the virulence and antibiotic resistance characteristics of the Salmonella strain, infections of varying severity could result. In this study, a multiplex melt curve real-time PCR assay for the detection of virulent and antibiotic resistance strains of Salmonella was developed with two primer sets. The first set targets the virulence gene, invasin (invA), and tetracycline (tetG), streptomycin (aadA2) and sulphonamide (sulI) antibiotic resistance genes, and the second set amplifies ampicillin (blaPSE,blaTEM) and chloramphenicol (floR) resistance genes. The multiplex assay was evaluated using 41 Salmonella strains and was further tested on eight different artificially inoculated food samples. The fluorescent DNA intercalating dye, SYTO9, generated high resolution melt curve peaks and, hence, was used for the development of the assay. This multiplex assay worked efficiently over a DNA concentration range of 20Â ng-200Â fg and showed a sensitivity of 290Â CFU/mL with serially diluted broth cultures. The detection limit for un-enriched artificially inoculated food samples was 104Â CFU/g, but an enrichment period of 6Â h allowed for detection of 10Â CFU/g of cells in the samples.
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Authors
Prashant Singh, Azlin Mustapha,