CHARACTERIZATION OF LEPTOSPIRA ISOLATES FROM SEROVAR HARDJO BY RIBOTYPING, ARBITRARILY PRIMED PCR, AND MAPPED RESTRICTION SITE POLYMORPHISMS

Leptospira serovar hardjo isolates of the hardjoprajitna and hardjobov is genotypes were characterized by ribotyping, arbitrarily primed PCR (AP-PCR) fingerprinting, and the study of mapped restriction site poly morphisms (MRSPs) in rrs and rrl genes. After restriction of chromosom al DNA with BglII, EcoRL, or HindIII, each genotype was individualized with a distinct ribotype. The fingerprints produced by AP-PCR with se ven primers clearly separated the two groups; primers KF and RSP produ ced species-specific products which assigned hardjoprajitno and hardjo bovis isolates to the species L. interrogans sensu stricto and L. borg petersenii, respectively. Furthermore, AP-PCR fingerprints gave eviden ce of a considerable genomic heterogeneity at the strain level among t he hardjobovis group. Conversely, the hardjoprajitno group was homogen eous. MRSP profiles in ribosomal genes indicated that hardjoprajitno a nd hardjobovis isolates belonged to L. interrogans MRSP group B and L. borgpetersenii group C, respectively. AP-PCR and determination of MRS Ps in ribosomal genes proved to be quick and reliable methods for typi ng Leptospira strains and for studying intraspecific population struct ures.