This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Martin, F. E.
Right arrow Articles by Hunter, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Martin, F. E.
Right arrow Articles by Hunter, N.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, May 2002, p. 1698-1704, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1698-1704.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Quantitative Microbiological Study of Human Carious Dentine by Culture and Real-Time PCR: Association of Anaerobes with Histopathological Changes in Chronic Pulpitis

F. Elizabeth Martin,* Mangala A. Nadkarni, Nicholas A. Jacques, and Neil Hunter

Institute of Dental Research, Westmead Centre for Oral Health, Westmead Hospital, Westmead, NSW, Australia

Received 14 December 2001/ Returned for modification 25 January 2002/ Accepted 12 February 2002

The bacteria found in carious dentine were correlated with the tissue response of the dental pulps of 65 teeth extracted from patients with advanced caries and pulpitis. Standardized homogenates of carious dentine were plated onto selective and nonselective media under anaerobic and microaerophilic conditions. In addition, real-time PCR was used to quantify the recovery of anaerobic bacteria. Primers and fluorogenic probes were designed to detect the total anaerobic microbial load, the genera Prevotella and Fusobacterium, and the species Prevotella melaninogenica, Porphyromonas endodontalis, Porphyromonas gingivalis, and Micromonas (formerly Peptostreptococcus) micros. The pulpal pathology was categorized according to the cellular response and degenerative changes. Analysis of cultured bacteria showed a predominance of gram-positive microorganisms, particularly lactobacilli. Gram-negative bacteria were also present in significant numbers with Prevotella spp., the most numerous anaerobic group cultured. Real-time PCR analysis indicated a greater microbial load than that determined by colony counting. The total number of anaerobes detected was 41-fold greater by real-time PCR than by colony counting, while the numbers of Prevotella and Fusobacterium spp. detected were 82- and 2.4-fold greater by real-time PCR than by colony counting, respectively. Real-time PCR also identified M. micros, P. endodontalis, and P. gingivalis in 71, 60, and 52% of carious samples, respectively. Correlation matrices of the real-time PCR data revealed significant positive associations between M. micros and P. endodontalis detection and inflammatory degeneration of pulpal tissues. These anaerobes have been strongly implicated in endodontic infections that occur as sequelae to carious pulpitis. Accordingly, the data suggest that the presence of high levels of these bacteria in carious lesions may be indicative of irreversible pulpal pathology.

* Corresponding author. Mailing address: Institute of Dental Research, Westmead Centre for Oral Health, Westmead Hospital, P.O. Box 533, Wentworthville NSW 2145, Australia. Phone: 61-2-9845-8763. Fax: 61-2-9845-7599. E-mail: efjelda{at}mail.usyd.edu.au.

Journal of Clinical Microbiology, May 2002, p. 1698-1704, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1698-1704.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Preza, D., Olsen, I., Aas, J. A., Willumsen, T., Grinde, B., Paster, B. J. (2008). Bacterial Profiles of Root Caries in Elderly Patients. J. Clin. Microbiol. 46: 2015-2021 [Abstract] [Full Text]  
  • Aas, J. A., Griffen, A. L., Dardis, S. R., Lee, A. M., Olsen, I., Dewhirst, F. E., Leys, E. J., Paster, B. J. (2008). Bacteria of Dental Caries in Primary and Permanent Teeth in Children and Young Adults. J. Clin. Microbiol. 46: 1407-1417 [Abstract] [Full Text]  
  • Hain, T., Otten, S., von Both, U., Chatterjee, S. S., Technow, U., Billion, A., Ghai, R., Mohamed, W., Domann, E., Chakraborty, T. (2008). Novel Bacterial Artificial Chromosome Vector pUvBBAC for Use in Studies of the Functional Genomics of Listeria spp.. Appl. Environ. Microbiol. 74: 1892-1901 [Abstract] [Full Text]  
  • Byun, R., Carlier, J.-P., Jacques, N. A., Marchandin, H., Hunter, N. (2007). Veillonella denticariosi sp. nov., isolated from human carious dentine. Int. J. Syst. Evol. Microbiol. 57: 2844-2848 [Abstract] [Full Text]  
  • Dalwai, F., Spratt, D. A., Pratten, J. (2007). Use of Quantitative PCR and Culture Methods To Characterize Ecological Flux in Bacterial Biofilms. J. Clin. Microbiol. 45: 3072-3076 [Abstract] [Full Text]  
  • Accetto, T., Avgustin, G. (2007). Studies on Prevotella nuclease using a system for the controlled expression of cloned genes in P. bryantii TC1-1. Microbiology 153: 2281-2288 [Abstract] [Full Text]  
  • Maskell, N. A., Batt, S., Hedley, E. L., Davies, C. W. H., Gillespie, S. H., Davies, R. J. O. (2006). The Bacteriology of Pleural Infection by Genetic and Standard Methods and Its Mortality Significance. Am. J. Respir. Crit. Care Med. 174: 817-823 [Abstract] [Full Text]  
  • Horz, H. P., Vianna, M. E., Gomes, B. P. F. A., Conrads, G. (2005). Evaluation of Universal Probes and Primer Sets for Assessing Total Bacterial Load in Clinical Samples: General Implications and Practical Use in Endodontic Antimicrobial Therapy. J. Clin. Microbiol. 43: 5332-5337 [Abstract] [Full Text]  
  • Suzuki, N., Yoshida, A., Nakano, Y. (2005). Quantitative Analysis of Multi-Species Oral Biofilms by TaqMan Real-Time PCR. Clin Med Res 3: 176-185 [Abstract] [Full Text]  
  • Chhour, K.-L., Nadkarni, M. A., Byun, R., Martin, F. E., Jacques, N. A., Hunter, N. (2005). Molecular Analysis of Microbial Diversity in Advanced Caries. J. Clin. Microbiol. 43: 843-849 [Abstract] [Full Text]  
  • Nadkarni, M. A., Caldon, C. E., Chhour, K.-L., Fisher, I. P., Martin, F. E., Jacques, N. A., Hunter, N. (2004). Carious Dentine Provides a Habitat for a Complex Array of Novel Prevotella-Like Bacteria. J. Clin. Microbiol. 42: 5238-5244 [Abstract] [Full Text]  
  • Suzuki, N., Nakano, Y., Yoshida, A., Yamashita, Y., Kiyoura, Y. (2004). Real-Time TaqMan PCR for Quantifying Oral Bacteria during Biofilm Formation. J. Clin. Microbiol. 42: 3827-3830 [Abstract] [Full Text]  
  • Nadkarni, M. A., Nguyen, K.-A., Chapple, C. C., DeCarlo, A. A., Jacques, N. A., Hunter, N. (2004). Distribution of Porphyromonas gingivalis Biotypes Defined by Alleles of the kgp (Lys-Gingipain) Gene. J. Clin. Microbiol. 42: 3873-3876 [Abstract] [Full Text]  
  • McLachlan, J. L., Sloan, A. J., Smith, A. J., Landini, G., Cooper, P. R. (2004). S100 and Cytokine Expression in Caries. Infect. Immun. 72: 4102-4108 [Abstract] [Full Text]  
  • Munson, M. A., Banerjee, A., Watson, T. F., Wade, W. G. (2004). Molecular Analysis of the Microflora Associated with Dental Caries. J. Clin. Microbiol. 42: 3023-3029 [Abstract] [Full Text]  
  • Byun, R., Nadkarni, M. A., Chhour, K.-L., Martin, F. E., Jacques, N. A., Hunter, N. (2004). Quantitative Analysis of Diverse Lactobacillus Species Present in Advanced Dental Caries. J. Clin. Microbiol. 42: 3128-3136 [Abstract] [Full Text]  
  • Boutaga, K., van Winkelhoff, A. J., Vandenbroucke-Grauls, C. M. J. E., Savelkoul, P. H. M. (2003). Comparison of Real-Time PCR and Culture for Detection of Porphyromonas gingivalis in Subgingival Plaque Samples. J. Clin. Microbiol. 41: 4950-4954 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»