• International Journal of Oral Biology
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• v.38 no.2
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• pp.43-49
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• 2013

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii-specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was assessed against 43 strains of mitis group streptococci, including clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amplicons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

• International Journal of Oral Biology
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• v.38 no.1
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• pp.29-36
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• 2013

Mitis group streptococci (MGS) were classified based on the nucleotide sequences 16S rRNA gene (16S rDNA) and comprised 13 Streptococcus species. However, 16S rDNA homogeneity among MGS was too high to discriminate between clinical strains at the species level, notably between Streptococcus mitis, Streptococcus oralis, Streptococcus pneumoniae, and Streptococcus pseudopneumoniae. The purpose of this study was to discriminate between 37 strains of MGS isolated from Korean oral cavities using phylogenetic analysis of the DNA-dependant RNA polymerase beta-subunit gene (rpoB). 16S rDNA and rpoB from clinical strains of MGS were sequenced using the dideoxy chain termination method and analyzed using MEGA version 5 software. The resulting phylogenetic data showed that the rpoB sequences could delineate clinical strains of MGS at the species level. Phylogenetic analysis of rpoB is therefore a useful approach for identifying MGS at the species level.

• International Journal of Oral Biology
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• v.31 no.1
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• pp.11-14
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• 2006

This study was undertaken to develop PCR primers for the identification and detection of Streptococcus anginosus using species-specific forward and reverse primers. These primers targeted the variable regions of the 16S ribosomal RNA coding gene(rDNA). The primer specificity was tested against 12 S. anginosus strains and 6 different species(10 strains) of oral bacteria. The primer sensitivity was determined by testing serial dilutions of the purified genomic DNA of S. anginosus ATCC $33397^T$. The data showed that species-specific amplicons were obtained from all the S. anginosus strains tested, but not in the six other species. The PCR could detect as little as 0.4pg of the chromosomal DNA from S. anginosus. This suggests that the PCR primers are highly sensitive and applicable to the detection and identification of S. anginosus.

• Journal of Microbiology
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• v.44 no.5
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• pp.572-576
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• 2006

To obtain primary idea on oral bacterium species that are generally present in periodotally healthy Koreans, the oral bacterial flora in the saliva of four periodontally healthy Koreans at different ages (5, 32, 35, 65) was investigated in this study. For this investigation, 16S rRNA gene clone libraries were generated from the saliva of the four healthy Koreans, and 50 clones were randomly selected from each saliva clone library and sequenced. Totally, 37 different kinds of bacterial 16S rRNA gene sequences were identified based on sequence homology search through GenBank database. The 37 kinds of saliva clone sequences were classified to 14 genera and 2 uncultured and 1 unidentified bacteria. Among the 14 identified genera, Streptococcus, Prevotella, and Veillollella were common genera, and Streptococcus was dominant genus that accounted for 7 different species. Among the seven Streptococcus species, S. salivarius appeared as the most common species. More numbers of species belonging to the genera Streptococcus and Prevotella was present in saliva from ages 32 and 35. While saliva from ages 5 and 65 showed more numbers of species belonging to the genera Rothia, including potential pathogenic species. Overall, saliva of a young child and a senior showed higher bacterial diversity than that of young adults.

• International Journal of Oral Biology
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• v.38 no.1
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• pp.21-27
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• 2013

Anginosus group streptococci (AGS) were classified based on the nucleotide sequences of the 16S rRNA gene (16S rDNA) and comprised Streptococcus anginosus, Streptococcus intermedius, and Streptococcus constellatus. It is known that AGS is a causative factor of oral and systematic diseases. The purpose of this study was to discriminate the 56 clinical strains of AGS isolated from Korean oral cavities using phylogenetic analysis of 16S rDNA and species-specific PCR at the species-level. The 16S rDNA of clinical strains of AGS was sequenced using the dideoxy chain termination method and analyzed using MEGA version 5 software. PCR was performed to identify the clinical strains using species-specific primers described in previous studies and S. intermedius-specific PCR primers developed in our laboratory. The resulting phylogenetic data showed that the 16S rDNA sequences can delineate the S. anginosus, S. intermedius, and S. constellatus strains even though the 16S rDNA sequence similarity between S. intermedius and S. constellatus is above 98%. The PCR data showed that each species-specific PCR primer pair could discriminate between clinical strains at the species-level through phylogenetic analysis of 16S rDNA nucleotide sequences. These results suggest that phylogenetic analysis of 16S rDNA and PCR are useful tools for discriminating between AGS strains at the species-level.

• International Journal of Oral Biology
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• v.35 no.1
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• pp.21-25
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• 2010

This study was undertaken to develop species-specific forward and universal reverse PCR primers for the detection of Streptococcus sobrinus. These primers target the variable regions of the 16S ribosomal RNA coding gene (rDNA) and their specificity was tested against 10 strains of S. sobrinus strains and 20 different species of oral bacteria using serial dilutions of the purified genomic DNA of S. sobrinus ATCC $33478^T$. Our data show that species-specific amplicons were obtained from all the S. sobrinus strains tested but not from other species. Both direct and nested PCR could detect as little as 400 pg and 4 fg of genomic DNA from S. sobrinus ATCC $33478^T$, respectively. This result suggests that these PCR primers are highly specific and sensitive and applicable to the detection of S. sobrinus.

• International Journal of Oral Biology
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• v.41 no.2
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• pp.75-81
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• 2016

Human mouth environment is known to include a variety bacteria, including Streptococcus spp., Staphylococcus spp., Actinomyces spp., Lactobacillus spp., Candida spp., Enterobacteriaceae, et al. Human oral microorganisms can cause dental caries, gingivitis, periodontitis, respiratory tract infection, and cardiovascular disease. Thus, right denture cleaning is essential to oral and general human health. The aim of this study was to evaluate the bactericidal effect of a sodium dichloroisocyanurate-based effervescent tablet (Aos Denti Germ, Aos Company, Chungbuk, Korea) against oral microorganisms. A total of 5 species Streptococcus spp. (Streptococcus anginosus, Streptococcus mitis, Streptococcus mutans, Streptococcus oralis, and Streptococcus sobrinus), Actinomyces oris, Candida albicans, and Escherichia coli were used in this study. All strains were exposed to the distilled water prepared with effervescent tablet. After the exposure, the mixture of strains and effervescent tablet was inoculated onto blood agar or MacConkey agar plate and cultured at $36^{\circ}C$. All strains were killed immediately on exposure to effervescent tablet. The results suggested that effervescent tablet could be used as an effective denture cleanser for dental hygiene.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.289-292
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• 2008

Trisodium phosphate 12 hydrate and citric acid monohydrate mixture showed the strong anti-sticking effect on Streptococcus mutans, Streptococcus mitis, and Streptococcus salivarius, which are adhered to glass beads. Each Streptococcus species was shaking-cultured in brain heart infusion broth containing three glass beads. After 18 hr, glass beads were slightly washed into normal saline by three-pin-pointed pincette. Each three glass-beads set was put into reagent -containing tubes, which have 40 mg of bits of weighing paper for gaining brushing effect as similar as brushing one's teeth. The tubes were shaken by vortex mixer for 10 min except non-oral microbe, Streptococcus agalactiae (5 min). The samples were colony-counted by serial agar dilution method. Experiment was repeated three times for each Streptococcus species. The relative ratios of bacterial de-adherence by reagents were calculated in comparison with normal saline control. The de-adherence degree of citric acid-trisodium phosphate-saline mixture (CTS, pH 6.0) against Streptococcus mutans came to an average of 12.5 times compared with normal saline control. Trisodium-saline (TS, pH 8.4) showed the average of 7.5 times, and citric acid-saline (CS, pH 4.6) showed 6.0 times compared to the control group. The bacterial de-adherence degree against Streptococcus salivarius was each 7.2,2.6 and 2.8 times in above reagent sequence in comparison with saline control. CTS and TS showed 2.4 and 3.4 times of anti-sticking effect on Streptococcus mitis respectively, but CS had no anti-sticking effect on this bacterium. CTS, TS and CS showed 0.7, 0.6, and 0.6 times on non-oral microbe, Streptococcus agalactiae, separately compared with saline control. These results show that oral Streptococcus mutans, Streptococcus salivarius, and Streptococcus mitis, which are causative of dental caries or subacute endocarditis, may be easily removed from oral cavity by CTS mixture. It is conceivable that our experimental results will enable the development of a new conceptive toothpaste to prevent dental caries or subacute endocarditis after drawing teeth.

• Korean Journal of Microbiology
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• v.53 no.2
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• pp.129-130
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• 2017

Streptococcus gordonii is a Gram-positive, facultative anaerobic and non-motile cocci. S. gordonii is a member of oral flora and a pioneer species that initiate the dental biofilm formation. S. gordonii has also been implicated in the pulpitis of primary teeth as well as systemic diseases such as infective endocarditis and septic arthritis. S. gordonii is associated with oral, respiratory, and gastrointestinal tract infections. S. gordonii KCOM 1506 (= ChDC B679) was isolated from a human acute pulpitis lesion. Here, we present the complete genome sequence of S. gordonii KCOM 1506.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.556-560
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• 2009

Xanthorrhizol, a sesquiterpene isolated from Curcuma xanthorrhiza Roxb., was used to investigate its effect on reducing the saliva and multi-species oral biofilms consisting of Streptococcus mutans, Streptococcus sanguis, and Actinomyces viscosus by anti-biofilm and confocal laser scanning microscopy (CLSM) assays. Xanthorrhizol exhibited significant antibiofilm activity in the dose- and time-dependent manners. Exposure to 2 and $5{\mu}g/mL$ xanthorrhizol for 30 min remained <50% of saliva and multi-species biofilms formed for 24 hr. In addition, exposure to $10{\mu}g/mL$ xanthorrhizol for 30 min reduced 65 and 77% of 24 hr saliva and multi-species oral biofilms, respectively. CLSM results visually demonstrated that xanthorrhizol reduced bacterial viability in the saliva and multi-species oral biofilms. These results suggest that C. xanthorrhiza Roxb. containing xanthorrhizol with strong anti-biofilm activity can be employed as a plant source for oral care functional foods.