Departmental Papers (Dental)

Document Type

Journal Article

Date of this Version

4-2011

Publication Source

Molecular Oral Microbiology

Volume

26

Issue

2

Start Page

127

Last Page

139

DOI

10.1111/j.2041-1014.2010.00603.x

Abstract

Approximately 35% of the species present in subgingival biofilms are as yet uncultivated, so their role in periodontal pathogenesis is unknown. The aim of the present study was to develop a high throughput method to quantify a wide range of cultivated and uncultivated taxa in subgingival biofilm samples associated with periodontal disease or health. Oligonucleotides targeting the 16S ribosomal DNA gene were designed, synthesized and labeled with digoxigenin. These probes were hybridized with the total nucleic acids of pure cultures or subgingival biofilm samples. Target species included cultivated taxa associated with periodontal health and disease, as well as uncultivated species, such as TM7 sp OT 346, Mitsuokella sp. OT 131 and Desulfobulbus sp. OT 041. Sensitivity and specificity of the probes were determined. A Universal probe was used to assess total bacterial load. Sequences complementary to the probes were used as standards for quantification. Chemiluminescent signals were visualized after film exposure or using a CCD camera. In a pilot clinical study, 266 subgingival plaque samples from eight periodontally healthy people and 11 patients with periodontitis were examined. Probes were specific and sensitivity reached 104 cells. Fusobacterium nucleatum ss polymorphum and Actinomyces gerencseriae were the most abundant cultivated taxa in clinical samples. Among uncultivated/unrecognized species, Mitsuokella sp. OT 131 and Prevotella sp. OT 306 were the most numerous. Porphyromonas gingivalis and Desulfobulbus sp. OT 041 were only detected in patients with periodontitis. Direct hybridization of total nucleic acids using oligonucleotide probes permitted the quantification of multiple cultivated and uncultivated taxa in mixed species biofilm samples.

Copyright/Permission Statement

This is the pre-peer reviewed version of the following article: [Teles, R. P., Teles, F. R. F., Loesche, W. J., Listgarten, M., Fine, D., Lindhe, J., . . . Sahlgrenska Academy. (2012). Rediscovering sig socransky, the genius and his legacy. Journal of Dental Research, 91(5), 433-439. doi:10.1177/0022034512443689], which has been published in final form at [http://dx.doi.org/10.1177/0022034512443689]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.

Comments

At the time of publication, author Flavia Teles was affiliated with the Harvard School of Dental Medicine. Currently, she is a faculty member at the School of Dental Medicine at the University of Pennsylvania.

Keywords

bacteria, biofilms, oral, periodontal, uncultivated

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Date Posted: 25 February 2022

This document has been peer reviewed.