Polymerase Chain Reaction-Restriction Fragment Length Polymorphism assay (PCR-RFLP) for porA and porB genes of Neisseria meningitidis was applied to type 64 strains isolated from cases of invasive disease in Italy. The technique was also successfully used on non-viable strains and on cerebrospinal fluid samples from patients with meningococcal meningitis diagnosed by PCR. RFLP patterns were obtained for all strains, including those nontypeable or nonsubtypeable by the serological methods. The results confirmed the usefulness of the PCR-RFLP for porA and porB genes of Neisseria meningitidis in differentiating strains belonging to the same serological type and provided discriminative patterns in the increased proportion of non serotypeable/serosubtypeable strains circulating in Italy in recent years.