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On the use of denaturing gradient gel electrophoresis approach for bacterial identification in endodontic infections

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Abstract

Bacteria in infected root canals of teeth evincing chronic apical periodontitis lesions were identified by a polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE) approach. DNA was extracted from root canal samples, and part of the 16S rRNA gene of all bacteria was amplified by PCR and separated by DGGE, generating banding patterns representative of the community structure. Twenty visible bands were cut out of the gel, re-amplified, and sequenced to provide identification. Sequencing analysis revealed the presence of both cultivable and as-yet-uncultivated species in the samples analyzed, including representatives of the genera Fusobacterium, Bacteroides, Dialister, Synergistes, Prevotella, Eubacterium and Peptostreptococcus. Unambiguous identification was not always possible and the method’s limitations are discussed. In general, the findings showed that PCR-DGGE can be useful for the identification of both cultivable and as-yet-uncultivated bacteria in endodontic infections.

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Acknowledgment

This study was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), a Brazilian governmental institution.

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Authors and Affiliations

  1. Universidade Estácio de Sá, Av. Paulo de Frontin 628, 5 andar, Rio de Janeiro, 20261-243, Brazil

    Julio C. Machado de Oliveira, Tulio G. V. Gama, José F. Siqueira Jr. & Isabela N. Rôças

  2. Institute of Microbiology Prof. Paulo de Góes, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    Raquel S. Peixoto & Alexandre S. Rosado

Authors
  1. Julio C. Machado de Oliveira
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  2. Tulio G. V. Gama
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  3. José F. Siqueira Jr.
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  4. Isabela N. Rôças
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  5. Raquel S. Peixoto
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  6. Alexandre S. Rosado
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Correspondence to José F. Siqueira Jr..

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Machado de Oliveira, J.C., Gama, T.G.V., Siqueira, J.F. et al. On the use of denaturing gradient gel electrophoresis approach for bacterial identification in endodontic infections. Clin Oral Invest 11, 127–132 (2007). https://doi.org/10.1007/s00784-006-0085-9

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  • DOI: https://doi.org/10.1007/s00784-006-0085-9

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