Using DGGE and 16S rRNA gene sequence analysis to evaluate changes in oral bacterial composition.
OBJECTIVE
To investigate whether a standard dental prophylaxis followed by tooth brushing with an antibacterial dentifrice will affect the oral bacterial community, as determined by denaturing gradient gel electrophoresis (DGGE) combined with 16S rRNA gene sequence analysis.


METHODS
Twenty-four healthy adults were instructed to brush their teeth using commercial dentifrice for 1 week during a washout period. An initial set of pooled supragingival plaque samples was collected from each participant at baseline (0 h) before prophylaxis treatment. The subjects were given a clinical examination and dental prophylaxis and asked to brush for 1 min with a dentifrice containing 0.3% triclosan, 2.0% PVM/MA copolymer and 0.243% sodium fluoride (Colgate Total). On the following day, a second set of pooled supragingival plaque samples (24 h) was collected. Total bacterial genomic DNA was isolated from the samples. Differences in the microbial composition before and after the prophylactic procedure and tooth brushing were assessed by comparing the DGGE profiles and 16S rRNA gene segments sequence analysis.


RESULTS
Two distinct clusters of DGGE profiles were found, suggesting that a shift in the microbial composition had occurred 24 h after the prophylaxis and brushing. A detailed sequencing analysis of 16S rRNA gene segments further identified 6 phyla and 29 genera, including known and unknown bacterial species. Importantly, an increase in bacterial diversity was observed after 24 h, including members of the Streptococcaceae family, Prevotella, Corynebacterium, TM7 and other commensal bacteria.


CONCLUSIONS
The results suggest that the use of a standard prophylaxis followed by the use of the dentifrice containing 0.3% triclosan, 2.0% PVM/MA copolymer and 0.243% sodium fluoride may promote a healthier composition within the oral bacterial community.
College of Dentistry, New York University, NY, USA.
Zhou
Z
Chen
Harsh M
HM
Trivedi
Nok
N
Chhun
Virginia M
VM
Barnes
Deepak
D
Saxena
Tao
T
Xu
Yihong
Y
Li
D000328:Adult; D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D000891:Anti-Infective Agents, Local / Q000627:therapeutic use; D001419:Bacteria / Q000145:classification* / Q000235:genetics; D002327:Cariostatic Agents / Q000627:therapeutic use; D003352:Corynebacterium / Q000145:classification; D004269:DNA, Bacterial / Q000032:analysis; D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods*; D003773:Dental Plaque / Q000382:microbiology*; D003777:Dental Prophylaxis; D003802:Dentifrices / Q000627:therapeutic use; D016680:Genome, Bacterial / Q000235:genetics; D006801:Humans; D008298:Maleates; D014677:Pharmaceutical Vehicles; D011095:Polyethylenes; D018720:Prevotella / Q000145:classification; D012329:RNA, Bacterial / Q000032:analysis*; D012336:RNA, Ribosomal, 16S / Q000032:analysis*; D017423:Sequence Analysis, RNA / Q000379:methods*; D012969:Sodium Fluoride / Q000627:therapeutic use; D013289:Streptococcaceae / Q000145:classification; D013997:Time Factors; D014099:Toothbrushing / Q000379:methods; D014260:Triclosan / Q000627:therapeutic use
D000900:Anti-Bacterial Agents; D000891:Anti-Infective Agents, Local; D002327:Cariostatic Agents; D004269:DNA, Bacterial; D003802:Dentifrices; D008298:Maleates; D014677:Pharmaceutical Vehicles; D011095:Polyethylenes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D014260:Triclosan; D012969:Sodium Fluoride; C043105:poly(methyl vinyl ether-co-maleic anhydride)