Real-time PCR methods for detecting Salmonella spp. in food after different DNA extraction procedures

Marko Dmitrić ,Kazimir Matović ,Dejan Vidanović ,Lj Saric,Nedjeljko Karabasil

doi:10.1088/1755-1315/333/1/012041

Marko Dmitrić , Kazimir Matović + Show 3 more

Open Access

https://doi.org/10.1088/1755-1315/333/1/012041

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Abstract

The aim of this paper was to evaluate two real-time PCR (qPCR) protocols for the detection of Salmonella spp. in minced meat and chicken neck skin, after DNA extraction using the InstaTM Gene matrix (BioRad, USA) and DNA extraction based on thermal cell lysis. The applied molecular methods were sensitive and specific for the rapid detection of Salmonella spp. in minced meat and chicken neck skin. The qualitative results were identical regardless of the applied DNA extraction or qPCR protocols. Lower Cq values were achieved after DNA extraction using the InstaTM Gene matrix.

Highlights

Salmonella species are one of the main foodborne pathogens [12]
The aim of this paper was to evaluate two real-time PCR protocols for the detection of Salmonella spp. in minced meat and chicken neck skin, after DNA extraction using the InstaTM Gene matrix (BioRad, USA) and DNA extraction based on thermal cell lysis
Lower Cq values were achieved after DNA extraction using the InstaTM Gene matrix

Summary

Introduction

Salmonella species are one of the main foodborne pathogens [12]. The most common sources of human infections are food products of animal origin, especially pork and poultry meat [3,4,5,6,7]. In the European Union, 91,662 cases of salmonellosis were confirmed during 2017 [14]. In Serbia, 1,850 cases of salmonellosis were diagnosed during 2017, which is 16.4% more cases than in 2016 [13]. The standard method requires at least four days for the detection of Salmonella spp. in food. Modern food microbiology demands the implementation of faster methods for the detection of Salmonella spp. The qPCR method meets this requirement, but it is still relatively more expensive than the cultural method. Evaluate a modified qPCR protocol for the detection of the invasion gene (inv A) [9] and the tetrathionate respiration gene (ttr) [11] Salmonella spp. in minced meat and chicken neck skin samples and to compare results with the reference method [17]. Compare two DNA extraction procedures and determine the effect of using different volumes of BPW pre-enrichments for the DNA extractions

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