Simultaneous detection of Salmonella pathogenicity island 2 and its antibiotic resistance genes from seafood

• Multiplex PCR assay for detecting virulence and antibiotic resistance is developed.
• Three virulence and three antibiotic resistance genes were targeted.
• The sensitivity of detection was confirmed in artificially contaminated samples.
• Efficiency of the assay was validated on 65 isolates of Salmonella from seafood.
• The assay can find application in seafood quality control and risk assessment.

Salmonella enterica serovars are virulent pathogens of humans and animals with many strains possessing multiple drug resistance traits. They have been found to carry resistance to ampicillin, chloramphenicol, florfenicol, streptomycin, sulfonamides, and tetracycline (ACSSuT-resistant). A rapid and sensitive multiplex PCR (mPCR)-based assay was developed for the detection of Salmonella serovars from seafood. Six sets of primers which are one primer pair targeting Salmonella specific gene invA (284 bp), two Salmonella pathogenicity island 2 (SPI-2) genes ssaT (780 bp) and sseF (888 bp) and three antibiotic resistance genes floR (198 bp), sul1 (425 bp), tetG (550 bp) were used for the study. The specificity and sensitivity of the assay were tested by spiking shrimp/fish/clam homogenate with viable cells of Salmonella. This assay allows for the cost effective and reliable detection of pathogenic Salmonella enterica from seafood. The mPCR developed in the present study proved to be a potent analytical tool for the rapid identification of multidrug-resistant Salmonella serovars from seafood.