Comparison of Multiple‑Locus Variable number tandem repeat Analysis and Pulsed Field Gel Electrophoresis in molecular subtyping of Listeria monocytogenes isolates from Italian cheese.

Subtyping of Listeria monocytogenes strains is an essential epidemiological tool to trace contamination and determine evolutionary relationships among different strains. Pulsed field gel electrophoresis (PFGE) is the current gold standard method for Listeria characterization. Multiple-Locus Variable-number tandem repeat Analysis (MLVA) is a rapid subtyping method based on Polymerase Chain Reaction (PCR) amplification that has been successfully developed for subtyping bacterial genera. The purpose of this study was to evaluate MLVA for subtyping L. monocytogenes strains isolated from Italian cheese and to compare it with PFGE. The type ability and discriminatory power of MLVA was determined on a collection of 90 isolates corresponding to 5 serotypes and 29 pulsotypes with enzymes AscI and ApaI. A panel of 5 variable-number tandem repeat loci was used. MLVA and PFGE showed very similar discriminatory power (Simpson's Index of diversity 0.840 with 95% Confidence Interval [CI] 0.780/0.899 and 0.837 with 95% CI 0.768/0.906, respectively). MLVA is an easy test to perform. It is relatively fast, reproducible and could be implemented in any molecular laboratory but, according to the performed protocol, it is not sufficient for discriminating the L. monocytogenes strains isolated from cheese. This method could be combined with PFGE to increase the discrimination in molecular subtyping of these strains.