3 SPECIES OF BORRELIA-BURGDORFERI SENSU LATE (BORRELIA-BURGDORFERI SENSU-STRICTO, B-AFZELII, AND B-GARINII) IDENTIFIED FROM CEREBROSPINAL-FLUID ISOLATES BY PULSED-FIELD GEL-ELECTROPHORESIS AND PCR

A total of 36 European Borrelia burgdorferi sensu lato cerebrospinal f luid isolates (mainly from southern Germany) were analyzed by pulsed-f ield gel electrophoresis (PFGE) for large restriction fragment pattern (LRFP) and linear plasmid profiles. Analyzing this large panel of iso lates, me detected all three species of B. burgdorferi sensu late path ogenic for humans in cerebrospinal fluid from patients with Lyme neuro borreliosis by PFGE typing after MluI digestion: 21 B. garinii (58%), 10 B. afzelii (28%), and 4 B. burgdorferi sensu stricto (11%) strains as well as 1 isolate with bands characteristic of both B. afzelii and B. garinii. Species classification by PFGE typing was confirmed by 16S rRNA-specific PCR. Eighteen isolates (11 B. garinii, 6 a B, afzelii, and 1 B. burgdorferi sensu stricto isolate) were further characterized by LRFP with four different restriction enzymes (ApaI, KspI, SmaI, an d KhoI). All B. afzelii isolates showed identical patterns for each re striction enzyme group. Considerable heterogeneity was demonstrated wi thin the B. garinii group. Subsequent analysis of plasmid profiles rev ealed only marginal differences for B. afzelii strains but different p atterns for B. garinii isolates. In one B. afzelii strain we found a l inear plasmid of about 110 kbp not described before. LRFP analysis by PFGE is a suitable tool for the molecular characterization of B. borgd orferi sensu late strains and allows determination not only of the spe cies but also of the subtypes within B. garinii.