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Abstract

MOLECULAR DETECTION OF ENTEROTOXIGENIC ISOLATES OF SALMONELLA TYPHIMURIUM, SHIGELLA FLEXNERI AND STAPHYLOCOCCUS AUREUS

C. Anish, M. Muthusamy and S. Ramesh*

ABSTRACT

The precise detection of pathogenic organisms that cause food-borne diseases is essential to ensure food safety. The diagnostic procedures should enable to detect enterotoxin producing these pathogens amidst other grown microbial flora and unknown complex food matrix. In the present study, we have developed multiplex PCR for detection of enterotoxigenic food borne pathogens such as Salmonella typhimurium, Shigella flexneri and Staphylococcus aureus by targeting toxin genes such as stn, shET and seB respectively. The efficiency of the mPCR method was confirmed with target-positive pure reference culture from spiked milk and bread samples. The detection of these pathogens by mPCR requires at least 36 ng of target-positive DNA from Staphylococcus aureus, 360 pg from Salmonella typhimurium and as low as 36 pg of DNA from Shigella flexneri in the presence of mixture of genomic DNA samples. The mPCR efficiency was evaluated in the presence of single target-positive and several other targetnegative Enterobacteriaceae family members. This method can be employed as molecular diagnostics method for detection of enterotoxigenic Salmonella typhimurium, Shigella flexneri and Staphylococcus aureus from food samples since mixture of DNAs isolated from spiked food samples provide equivalent efficiency in pathogen detections.

Keywords: Genomic DNA, mPCR, enterotoxin gene, food-borne pathogens.


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