Application of Real-time PCR for Rapid Petection of Salmonella spp., Salmonella enterica ser. Typhimurium and Enteritidis in Food Samples of Animal Origin without Pre-enrichment and with Pre-enrichment

Abstract

The aim of this study was follow the contamination of milk and meat products with Salmonella spp. by using the Step One real-time PCR. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and Power SYBR Green PCR Master Mix for pursuance the real-time PCR (Applied Biosystems). We were observing the presence of genes stn (Salmonella enterica ser. Typhimurium DT096), sef and pef (Salmonella enterica ser. Enteritidis SE7). We could detect strain of Salmonella spp. in the investigated samples without pre-enrichment and after pre-enrichment. Our results indicated that the Step One real-time PCR assay developed in this study could sensitively detect Salmonella spp. in food samples of animal origin (swabs). This Step One real-time PCR assay is extremely useful for any laboratory in possession of a real-time PCR. It is a fast, reproducible, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future. Our results indicated that the Step One real-time PCR assay developed in this study could sensitively detect Salmonella spp. in ready to eat food. Thus, these results proved real-time PCR to be useful as a rapid diagnostic test for the direct detection of pathogens in food of animal origin, without the need of pre-enrichment steps.

Keywords

• pre-enrichment
• real-time pcr
• salmonella spp.
• sybr green