A low-cost, rapid, sensitive and reliable PCR-based alternative method for predicting the presence of possible live microbial contaminants in food

In this study we report the development of a cheaper, faster and more sensitive alternative method than the commonly used RT-PCR, and more reliable than conventional DNA-based PCR method for the prediction of the presence of possible live pathogens in food. The false-positivity associated with the conventional PCR methods could be completely overcome by incorporating simple modifications at a step prior to the isolation of the template DNA from bacterial cells. We also report the development of a highly specific four-gene targeted multiplex PCR assay which would be useful for the simultaneous detection of two of the most important food-borne human pathogens, E. coli O157:H7 and L. monocytogenes.