IDENTIFICATION AND STRAIN DIFFERENTIATION OF BACTEROIDES-FRAGILIS GROUP SPECIES AND PREVOTELLA-BIVIA BY PCR FINGERPRINTING

Using single consensus primers of genomic nucleotide sequences, PCR-ge nerated fingerprints were used for identification and differentiation of the Bacteroides fragilis group (B, fragilis, B. thetaiotaomicron, B . ovatus, B, distasonis, B, vulgatus) and Prevotella bivia (B. bivius) by comparing the DNA profiles with those of reference strains from th e American Type Culture Collection and German Culture Collection. When primed by a single primer phage M13 core sequence, intra-species spec ific differences and species-specific bands were detected. Using prime rs derived from the evolutionarily conserved tRNA gene sequence, speci es-specific patterns were produced. A computer program, GelManager, wa s used to analyze the profiles and generate dendrograms. The correlati on coefficients determined from the DNA fingerprint profiles of the cl inical isolates (using the M13 core primer) fell within a narrow range , reflecting a high level of homology within the species. Based on the dendrograms, strains of one species were clearly differentiated from strains of other species. For comparison, SDS-PAGE analysis of whore c ell extracts was also performed to obtain protein band patterns of var ious strains. Because of the simplicity of the PCR fingerprinting meth od and the ease of performance of computerized evaluation of data, thi s technique is a useful method for both species and strain differentia tion, as well as for characterization of Bacteroides species and Prevo tella bivia. (C) 1995 Academic Press