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Journal of Clinical Microbiology, September 2007, p. 3039-3049, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.02618-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Use of Multiple-Displacement Amplification and Checkerboard DNA-DNA Hybridization To Examine the Microbiota of Endodontic Infections{triangledown}

L. C. N. Brito,2* F. R. Teles,1,3 R. P. Teles,1 E. C. França,2 A. P. Ribeiro-Sobrinho,2 A. D. Haffajee,1 and S. S. Socransky1

Department of Periodontology, The Forsyth Institute, Boston Massachusetts,1 Federal University of Minas Gerais School of Dentistry, Belo Horizonte, Minas Gerais, Brazil,2 Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, Boston, Massachusetts3

Received 31 December 2006/ Returned for modification 23 February 2007/ Accepted 6 July 2007

Multiple-displacement amplification (MDA) has been used to uniformly amplify bacterial genomes present in small samples, providing abundant targets for molecular analysis. The purpose of this investigation was to combine MDA and checkerboard DNA-DNA hybridization to examine the microbiota of endodontic infections. Sixty-six samples were collected from teeth with endodontic infections. Nonamplified and amplified samples were analyzed by checkerboard DNA-DNA hybridization for levels and proportions of 77 bacterial taxa. Counts, percentages of DNA probe counts, and percentages of teeth colonized for each species in amplified and nonamplified samples were computed. Significance of differences for each species between amplified and nonamplified samples was sought with Wilcoxon signed-rank test and adjusted for multiple comparisons. The amount of DNA in the samples ranged from 6.80 (± 5.2) ng before to 6.26 (± 1.73) µg after MDA. Seventy of the 77 DNA probes hybridized with one or more of the nonamplified samples. All probes hybridized with at least one sample after amplification. Most commonly detected species at levels of >104 in both amplified and nonamplified samples were Prevotella tannerae and Acinetobacter baumannii at frequencies between 89 and 100% of samples. The mean number of species at counts of >104 in amplified samples was 51.2 ± 2.2 and in nonamplified samples was 14.5 ± 1.7. The endodontic microbiota was far more complex than previously shown, although microbial profiles at teeth with or without periradicular lesions did not differ significantly. Species commonly detected in endodontic samples included P. tannerae, Prevotella oris, and A. baumannii.

* Corresponding author. Mailing address: Federal University of Minas Gerais School of Dentistry, Av. Antônio Carlos, 6627-Sl 3310 Bairro: Pampulha Cep:31270-901, Belo Horizonte, MG Brazil. Phone: (55)-31-3441-3915. Fax: (55)-31-3418-1433. E-mail: luitauna{at}yahoo.com.br

{triangledown} Published ahead of print on 18 July 2007.

Journal of Clinical Microbiology, September 2007, p. 3039-3049, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.02618-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Rocas, I. N., Siqueira, J. F. Jr. (2008). Root Canal Microbiota of Teeth with Chronic Apical Periodontitis. J. Clin. Microbiol. 46: 3599-3606 [Abstract] [Full Text]  


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