Quantification of Salmonella spp., Listeria monocytogenes and Escherichia coli O157:H7 in non-spiked food products and evaluation of real-time PCR as a diagnostic tool in routine food analysis

Escherichia coli O157:H7, Listeria monocytogenes and Salmonella spp. are foodborne pathogens frequently associated with foods such as poultry, ready-to-eat products, fruits and vegetables. PCR-based procedures are rapid, sensitive and accurate; in particular, real-time PCR (qPCR), which besides being an automated high-throughput technique, allows quantification of foodborne pathogens. In the present work, qPCR-based methods were applied for the quantitative detection of E. coli O157:H7, Salmonella spp. and L. monocytogenes in a total of 306 non-spiked food samples in a study carried out in two laboratories simultaneously. qPCR allowed the detection of the three pathogens in around 20% of the analyzed samples for each pathogen. Quantification results revealed the presence of the three pathogens mostly at levels between 102 and 104 cells/g. Besides quantification, the qPCR results (presence/absence) were compared with those of the standard mini-VIDAS system. In order to determine which were the “true” positive samples, conventional PCR was carried out after the corresponding enrichment for each pathogen. These results were considered as the gold standard for further analysis. The statistical analysis of global data recording the presence of E. coli O157:H7 and L. monocytogenes, together with data previously obtained for Salmonella spp., revealed that qPCR outperformed the mini-VIDAS procedures, in terms of both time and accuracy. Thus, these results proved qPCR to be useful as a rapid diagnostic test for the direct detection of pathogens in food, without the need for enrichment steps.