BROAD-RANGE PCR AMPLIFICATION AND DNA-SEQUENCE ANALYSIS REVEALS VARIABLE MOTIFS IN 16S RIBOSOMAL-RNA GENES OF MOBILUNCUS SPECIES

Using DNA primers based on highly conserved regions of bacterial 16S r ibosomal RNA genes, a technique was established for detection of Mobil uncus species by polymerase chain reaction (PCR) and hybridization ana lysis. Part of the 16S rRNA genes of Mobiluncus mulieris, Mobliuncus c urtisii and uncharacterized Mobiluncus strains were analyzed by broad- range PCR amplification and direct DNA sequencing analysis. Sequence c omparison of the partial 16S rRNA genes of Mobiluncus curtisii, Mobilu ncus mulieris and atypical Mobiluncus strains studied indicated genus and species-specific motifs within the variable regions V3, V4 and V9 of 16S ribosomal DNAs. A Mobiluncus curtisii-specific primer, located within the variable region V3 of the 16S rRNA gene, was designed for S outhern blot hybridization analysis of broad-range PCR products. Broad -range amplification combined with a M. curtisii-specific hybridizatio n probe, Mob V3, distinguished between Mobiluncus curtisii, Mobiluncus mulieris, and atypical Mobiluncus strains.