A Multiplex PCR Assay for Detection of Salmonella sp., Listeria monocytogenes, Campylobacter sp., Staphylococcus aureus and Escherichia coli

Contaminated food and water are usual vehicles for bacterial pathogens transmission. According to EFSA they promote foodborne illness. Salmonella sp., Campylobacter sp., Listeria monocytogenes, Escherichia coli and Staphylococcus aureus are the most foodborne pathogens reported. Bacterial contamination levels remain at high level, particularly in Europe, despite regulatory efforts to address the situation. The need of new diagnostic tools is crucial. Ideally, tests easy to perform, enough accurate and low cost. The present work talk about optimization of a multiplex PCR (mPCR) test used to detect 5 foodborne contaminants: Salmonella sp., Campylobacter sp., L. monocytogenes, E. coli and S. aureus. For specificity evaluation, 5 PCR amplification reference DNA were used respectively: 103bp, 174bp, 151bp, 121bp and 136bp. No amplification was observed when primers and DNA from mismatching species were subject to PCR amplification. Furthermore, the sensitivity of this assay was evaluated by using serial dilutions of DNA extracted from clean 1CFU culture of each pathogen. This assay will be optimized by using Real-Time PCR and DNA plasmids containing a single copy of each gene, towards a new and rapid test for food and food manipulated surfaces control. Results are promising and allow us to postulate the design of an accurate and useful assay for bacterial control.