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Increase in Probing Depth Is Correlated With a Higher Number of Prevotella intermedia Genotypes
Author(s) -
Alves Ana Cláudia B.A.,
Napimoga Marcelo H.,
Klein Marlise I.,
Hofling Jose F.,
Gonçalves Reginaldo B.
Publication year - 2006
Publication title -
journal of periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.036
H-Index - 156
eISSN - 1943-3670
pISSN - 0022-3492
DOI - 10.1902/jop.2006.77.1.61
Subject(s) - heteroduplex , prevotella intermedia , genotype , biology , amplicon , polymerase chain reaction , intermedia , genetic diversity , microbiology and biotechnology , genetics , dna , gene , medicine , bacteria , population , art , environmental health , porphyromonas gingivalis , performance art , art history
Background: The aims of this study were to determine the genotypic diversity of Prevotella intermedia in subgingival plaque samples by using two techniques, arbitrarily primed polymerase chain reaction (AP‐PCR) and heteroduplex analysis, and to assess the relationship of this diversity with increase in probing depth. Methods: The subgingival plaque samples were obtained from 12 patients using paper points inserted into periodontal pockets (diseased sites) and healthy gingival sulci (healthy sites) of the same subjects. After isolation and identification, AP‐PCR was performed for genotypic characterization of P. intermedia (80 isolates). The clinical samples with a positive result for P. intermedia were amplified by 16S rRNA‐based PCR method, and the amplicons were subjected to heteroduplex analysis. Results: The agreement between the two methods was very high; the AP‐PCR and heteroduplex analysis showed that subjects harbored between one and five distinct genotypes of P. intermedia , with a positive association between numbers of genotypes by AP‐PCR ( P = 0.0042) or heteroduplex ( P = 0.0099) and increase in probing depth. No matching of P. intermedia genotypes was observed between healthy and diseased sites of the same individual. Interindividual analyses demonstrated absence of identical clones and indicated a high level of genetic diversity in the species. Conclusion: A clear relationship was observed between a higher number of genotypes and increase in probing depth; these results suggest that environmental challenges in the periodontal pockets may modulate the microbiota by selecting genotypes best able to exploit the environment.