Mitis-salivarius sucrose bacitracin(MSB) medium is widely used in the selective isolation of mutans streptococci(MS), a designation for a group of oral cariogenic species. Recently, we have isolated three bacterial strains grown on MSB agar from human dental plaques. The three strains exhibited bioc...
Mitis-salivarius sucrose bacitracin(MSB) medium is widely used in the selective isolation of mutans streptococci(MS), a designation for a group of oral cariogenic species. Recently, we have isolated three bacterial strains grown on MSB agar from human dental plaques. The three strains exhibited biochemical characteristics similar to those of the biotype IV of MS, with the exception that they manifested a positive reaction for arginine deaminase. The objective of this study was to identify and characterize these three clinical isolates. The bacteria were identified with biochemical tests as well as by 16S rDNA cloning and sequencing. In order to compare the antibiotics susceptibility of the clinical isolates with that of type strain, the minimum inhibitory concentrations of 9 antibiotics were determined using broth dilution assays. The results identified all of our three clinical isolates as Enterococcus faecalis. All E. faecalis strains were found to be susceptible to penicillin G, amoxicillin, augmentin, and vancomycin, but were resistant to ciprofloxacin, cefuroxim axetil, and clindamycin. Our findings indicate that E. faecalis is capable of growing on MSB agar, and suggest that the MSB medium be improved so that only MS should be recoverable on the medium, as originally devised for their selection.
Mitis-salivarius sucrose bacitracin(MSB) medium is widely used in the selective isolation of mutans streptococci(MS), a designation for a group of oral cariogenic species. Recently, we have isolated three bacterial strains grown on MSB agar from human dental plaques. The three strains exhibited biochemical characteristics similar to those of the biotype IV of MS, with the exception that they manifested a positive reaction for arginine deaminase. The objective of this study was to identify and characterize these three clinical isolates. The bacteria were identified with biochemical tests as well as by 16S rDNA cloning and sequencing. In order to compare the antibiotics susceptibility of the clinical isolates with that of type strain, the minimum inhibitory concentrations of 9 antibiotics were determined using broth dilution assays. The results identified all of our three clinical isolates as Enterococcus faecalis. All E. faecalis strains were found to be susceptible to penicillin G, amoxicillin, augmentin, and vancomycin, but were resistant to ciprofloxacin, cefuroxim axetil, and clindamycin. Our findings indicate that E. faecalis is capable of growing on MSB agar, and suggest that the MSB medium be improved so that only MS should be recoverable on the medium, as originally devised for their selection.
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제안 방법
sobrinus) of mutans streptococci, with the exception that our isolates manifested a positive reaction for arginine deaminase. Therefore, in this study, we performed 16S rDNA sequencing and biochemical test using API 20 STREP (BioMerieux, Marcy 1'Etoile, France) in order to identify the isolates at the species level. In addition, antibiotic suscep tibility of the bacteria was eval니ated in order to characterize the bacteria.
To confirm the result of identification of the three strains by 16S rDNA sequence comparison method, biochemical tests were performed using commercial identification kit, API 20 STREP (BioMerieux, Marcy 1'Etoile, France) according to the manufacturer's instruction.
대상 데이터
E.faecalis KCTC 3206T (ATCC 19433T) and S. sobrinus ATCC 33478 T were obtained from the Korean Collection for Type Cultures (KCTC, Daejeon, Korea) and American Type C니ture Collection (ATCC, University Boulevand, Manassas, 、VA, USA), respectively. The clinically isolated strains, E.
이론/모형
, Madison, WI, USA). The nucleotide sequencing of the 16S rDNA was determined by use of the dideoxy chain termination method, with a Big Dye Terminator Cycle Seq니encing Kit (Applied Biosystems, Foster City, CA, USA) and an ABI PRISM 310 Genetic Analyzer (Applied Biosystems). The primers used in nucleotide sequencing were as follows: ChDC-GEM-F (5'-TTC CCA GTC ACG ACG TTG TAA AA-3'), Seq-Fl X 七CCT ACG GGA GGC AGC AG-39, Seq-R2 (5、GAC TAC CAG GGT ATC TAA TCC-3'), F16 (5'-TAG ATA CCC YGG TAG TCC-3, ), and ChDC-GEM-R (5'-GTG TGG AAT TGT GAG CGG ATA AC-3').
성능/효과
faecalis. The 16S rDNA sequences of the three clinical isolates were highly homologous (>98%) with that (GenBank accession number AB 154827) of E. faecalis EC-12 (Fig. 1). The GenBank accession numbers of 16S rDNA sequences derived from E.
1). The GenBank accession numbers of 16S rDNA sequences derived from E. faecalis ChDC YE1, ChDC YE2, and ChDC YE3 were AY942557, AY942558, and AY942559, respectively. The biological codes of API 20 STREP test for the three strains were 5173711.
, 2005). The results indicated that we had succeeded in isolating such organisms as S. anginosus, S. sanguinis, and Pantoea agglomerans.
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