Although isolates of the "Streptococcus milleri" group (SMG) of bacteria are regarded as members of the commensal microflora of the body, they are frequently encountered in purulent infections from a range of body sites. The genetic diversity of 91 epidemiologically unrelated SMG isolates (including 37 commensal strains and 49 disease-associated strains) was analyzed by macrorestriction fingerprinting (MF). The genomes were digested with SmaI and ApaI independently, and fragments were resolved by pulsed-field gel electrophoresis. Similarities between banding profiles were determined, and strains were clustered on this basis into dendrograms. In common with other commensal species that have been examined by MF, considerable genetic diversity was revealed. In addition, the clustering of strains tended to support the current taxonomic position of this heterogeneous group. The present study has shown that MF is a powerful tool for characterization of SMG strains and that its use is likely to be of great value in epidemiological and population genetic studies of this group of bacteria.
MATERIALS AND METHODSTest strains. The isolates of SMG were obtained from dentoalveolar abscesses (n ϭ 21), extraoral infections (n ϭ 28), and healthy oral sites (n ϭ 37); and 5 isolates were of unknown origin. Strains isolated from extraoral sites originated from blood, brain, lung, abdominal, perianal, urogenital, skin, soft tissue, bone, and miscellaneous infections ( Table 1). The majority of the nonoral isolates were kindly provided by R. Whiley (Oral Microbiology, St. Bartholomew's and the Royal London, United Kingdom). Commensal oral strains of SMG (Table 2) were isolated from the mixed saliva, supra-and subgingival plaques, dorsum of the tongue, and throat swabs obtained from unrelated healthy individuals and were cultured by use of a selective medium (4). The isolates were routinely maintained either in brain heart infusion broth (Oxoid Ltd., Basingstoke, United Kingdom) or on blood base agar (Oxoid Ltd.) containing 5% (vol/vol) defibrinated horse blood (TCS Microbiology, Botolph Claydon, United Kingdom). Cultures were incubated at 37°C in an anaerobic workstation (Don Whitley Scientific Ltd., Shipley, United Kingdom) containing an atmosphere of 10% hydrogen, 10% carbon dioxide, 80% nitrogen.A total of 91 clinical SMG isolates were examined in the study, including 3 type strains, S. anginosus NCTC 10713 (ATCC 12395); S. constellatus NCTC 11325 (ATCC 27823), and S. intermedius NCTC 11324 (ATCC 27335) (Table 1). Each strain was assigned to either S. anginosus, S. constellatus, or S. intermedius according to a differential phenotypic identification scheme (37). Briefly, strains were tested for a panel of six glycosidase reactions by using 4-methylumbelliferyllinked fluorogenic substrates and hyaluronidase production by a modified plate assay (37). The identity of each isolate was confirmed with the Rapid ID 32 Strep API system (bioMérieux sa, Marcy l'Etoile, France). All isolates were tested for the presence of Lancefield group an...