Optimization of a Viability PCR Method for the Detection of Listeria monocytogenes in Food Samples
Rapid detection of Listeria and other microbial pathogens in food is an essential part of quality control and it is critical for ensuring the safety of consumers. Culture-based methods for detecting foodborne pathogens are time-consuming, laborious and cannot detect viable but non-culturable microorganism, whereas viability PCR methodology provides quick results; it is able to detect viable but non-culturable cells, and allows for easier handling of large amount of samples. Although the most critical point to use viability PCR technique is achieving the complete exclusion of dead cell amplification signals, many improvements are being introduced to overcome this. In the present work, the yield of dead cell DNA neutralization was enhanced by incorporating two new sample treatment strategies: tube change combined with a double light treatment. This procedure was successfully tested using artificially contaminated food samples, showing improved neutralization of dead cell DNA.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 4.D’Agostino M, Wagner M, Vazquez-Boland JA, Kuchta T, Kapriskova R, Hoorfar J, Novella S, Scortti M, Ellison J, Murray A, Fernandes I, Kuhn M, Pazlarova J, Heuvelink A, Cook N (2004) A validated PCR-based method to detect Listeria monocytogenes using raw milk as a food model-towards an international standard. J Food Prot 67:1646–1655CrossRefPubMedGoogle Scholar
- 7.FDA (2015) Testing methodology for Listeria species or L. monocytogenes in Environmental Samples, version 1Google Scholar
- 21.UNE-EN ISO 11290-1 (2017) Microbiology of the food chain-Horizontal method for the detection and enumeration of Listeria monocytogenes and other Listeria spp.-Part 1: detection methodGoogle Scholar
- 22.UNE-EN ISO 7218:2007-Microbiology of food and animal feeding stuffs-general requirements and guidance for microbiological examinationsGoogle Scholar
- 23.UNE-EN ISO 16140-2:2016-Microbiology of the food chain-Method validation. Part 2: protocol for the validation of alternative (proprietary) methods against a reference methodGoogle Scholar