Abstract

One of the most deadly foodborne pathogens is the bacterium Listeria monocytogenes (L. monocytogenes). Though L. monocytogenes has a low incidence, its mortality rate is dangerously high. Because of the threat of this pathogen, President Clinton issued a directive that specifically called for a more accurate and sensitive detection protocol of L. monocytogenes. Using the portions of the hlyA (hemolysin A) gene sequence from L. monocytogenes, I designed and conducted experiments to find a primer/probe combination that would both sensitively and specifically detect L. monocytogenes using a real-time Polymerase Chain Reaction 5' nuclease assay (real-time PCR) as compared to traditional culturing methods and traditional PCR. I worked with various strains of L. monocytogenes, other Listeria species, as well as non-Listeria strains to determine the sensitivity and specificity of my primer/probe set and the real-time PCR assay itself. The results successfully exemplified the ability of the real-time PCR assay to be specific, rapid, simple, and reproducible for the detection of L. monocytogenes with the primer/probe combinations I developed.