Denaturing gradient gel electrophoresis analysis to study bacterial community structure in pockets of periodontitis patients.
Bacteria are involved in the onset and progression of periodontitis. A promising molecular technique, denaturing gradient gel electrophoresis (DGGE), to study microbial population dynamics in the subgingival pocket is presented. Twenty-three samples were taken from the subgingival pockets of nine patients and six healthy family members. From four periodontitis patients, 12 samples were evaluated before, 1 day after and 3 months after treatment. Part of the 16S rRNA gene of all bacteria was amplified by PCR and separated by DGGE, creating banding patterns representative of the community structure. Shifts in composition and diversity of the microbial population could be determined semiquantitatively, and this showed that treatment resulted in a decrease in the diversity of the population. After 3 months a microbial population 33-47% different from the population before treatment had re-established. Intense bands representing Exiguobacterium aurantiacum were present in 13 out of 25 samples, indicating that this species may play a role in periodontal disease.
Zijnge
V
V
Department of Medical Microbiology, University of Groningen, Groningen, the Netherlands.
Harmsen
H J M
HJ
Kleinfelder
J W
JW
van der Rest
M E
ME
Degener
J E
JE
Welling
G W
GW
D000190:Actinomyces / Q000235:genetics; D000328:Adult; D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics; D001421:Bacteria, Anaerobic / Q000235:genetics; D015373:Bacterial Typing Techniques; D018441:Biofilms / Q000254:growth & development*; D016022:Case-Control Studies; D004269:DNA, Bacterial / Q000032:analysis; D003773:Dental Plaque / Q000382:microbiology; D012534:Dental Scaling; D017753:Ecosystem; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005673:Fusobacterium / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010514:Periodontal Pocket / Q000382:microbiology* / Q000628:therapy; D016133:Polymerase Chain Reaction; D011489:Protein Denaturation; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S