Serogroup B has been the main cause of meningococcal disease in Spain since at least 1979, but in recent years an increase in the prevalence of infection due to serogroup C meningococci has been detected. In 1996, for the first time, most cases of meningococcal disease were caused by serogroup C strains. The sero/subtype of all serogroup C meningococci received from 1993 to June 1996 was determined, and the results showed that C:2b:P1.2,5, the most common phenotype in 1995 and 1996 (63% and 65%, respectively), represented only 4.8% of strains in 1993. The C:2b: P1.2,5 epidemic strains appear to be responsible for the high prevalence of serogroup C in Spain. One hundred fifty-one randomly selected serogroup C strains were analyzed by multilocus enzyme electrophoresis, ribotyping, and pulsed-field gel electrophoresis. Pulsed-field gel electrophoresis provided the most accurate information: more than 80% of the C:2b:P1.2,5 and C:2b:P1.2 isolates exhibited one of two very closely related profiles, while most of the C:2b:NST and C:2b:P1.5 strains had a pattern located at a genetic distance of 0.24 from those two profiles. The results show that C:2b:P1.2,5 strains represent a subclone or a genetic variant of the previously identified Spanish epidemic clone C:2b:non-subtypable strains.