The validation of the assay was performed with laboratory strains and clinical isolates. EGD and LO28, 11 serogroup 3 strains and ten strains of other serogroups for developing the molecular typing assay. The present study was conducted using two different type strains of L. monocytogenes strains that had been previously characterized by pulsed-field gel electrophoresis. Nucleotide sequence alignment and molecular modelling were used to identify regions of listeriolysin O gene of 37 strains of L. The purpose of the present study was to develop a novel PCR-based method for the molecular typing of Listeria monocytogenes strains. Among the different methods available, amplification of the listeriolysin O gene (hlyA) is the best characterized. In this scenario, typing methods are essential to better understand the epidemiology of the disease in the recent years. The number of reported cases of listeriosis has been increasing in recent years and new outbreaks have been reported in Italy in the past few years. The most common source of foodborne listeriosis is the consumption of contaminated food. Rapid diagnosis of patients with listeriosis is crucial for an appropriate treatment and also for contact tracing. Infections associated with food-borne listeriosis are a severe and life-threatening foodborne disease that mainly affect immunocompromised and elderly patients. Of a targeted diagnostic assay for typing of Listeria monocytogenes. It integrates well with the browser and is very easy to configure.
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