Listeria monocytogenes is a foodborne pathogen frequently present in ripened soft cheeses. Forty-three strains of L. monocytogenes isolated from the rind of ripened Gorgonzola cheeses produced in 24 different dairy plants were characterized by biotyping, serotyping, and molecular typing. Biotyping was performed by studying two phenotypes closely associated with virulence, such as hemolytic and phospholipase C activities. Traditional typing techniques did not allow a discrimination among the 43 strains studied. All strains showed a good hemolytic activity on blood agar, and only slight differences were observed when titration of hemolytic activity of culture supernatants was performed. Also phospholipase activities were quite similar for all the strains. Concerning serotyping, all strains belonged to serotype 1/2a. The molecular characterization was performed by RAPD-PCR. Combined cluster analysis following PCR amplification experiments allowed to group L. monocytogenes strains into few distinguishable profiles. At a level of similarity of 80%, the 43 strains were grouped into only 5 composite profile groups. Although isolated in 24 different plants, the presence of a few closely related strains demonstrated a possible relationship between these cheese isolates; a special ability of these strains to adapt to Gorgonzola cheese processing environment could be suggested.