Technical report on optimization of a reduced PCR reaction volume and a single PCR condition for five different foodborne pathogens

Abstract

Foodborne pathogens have caused many disease outbreaks globally and more so in developing countries with huge economic effect. Food-borne pathogen identification is an important aspect of health care. The short shelf-life of some food products such as vegetables requires rapid and cost-effective methods for pathogen detection. Therefore this work was conducted to optimize a reduced PCR reaction volume and a single PCR condition that can simultaneously be used to detect five different foodborne pathogens namely Salmonella spp, Escherichia coli (0157), Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus. After analyzing five protocols from literature which were not able to successfully amplify the target genes of the organisms simultaneously, an in-house method was developed which successfully amplified the genes of the five organisms. The findings of this study suggests that target genes of multiple organisms can be amplified within the shortest period of time using this newly developed in-house method. This approach will also decrease the cost of performing PCR analyses for pathogen identification particularly in developing countries