Identification of Actinobacillus actinomycetemcomitans in subgingival plaque by PCR.

Journal of Clinical Microbiology
T F FlemmigH Karch

Abstract

The purpose of this study was to assess the sensitivity and specificity of the PCR in detecting Actinobacillus actinomycetemcomitans. The PCR's detection capability was compared with those of three other methods: culture-enhanced PCR (CE-PCR), colony hybridization (CH), and conventional culture with presumptive biochemical identification. A 285-bp stretch of the leukotoxin gene lktA of A. actinomycetemcomitans was amplified by PCR with primers TT-15 and TT-16. For CH, the PCR product was labeled with digoxigenin and used as a hybridization probe. Nucleotide sequence analysis of the PCR product of A. actinomycetemcomitans 1D4 and 1664 and three clinical isolates revealed complete homology among the tested strains, with only one base substitution (at position 1344) in comparison with the published sequence. With artificially infected subgingival plaque, the detection limit of PCR for A. actinomycetemcomitans was 10(3) CFU/ml of plaque suspension. Culturing subgingival plaque on tryptic soy-serum-bacitracin-vancomycin agar prior to PCR (CE-PCR) improved the limit of detection to 10(2) CFU/ml. Analysis of subgingival plaque samples from 35 patients with periodontal disease and 10 periodontally healthy subjects revealed that CE-PCR ...Continue Reading

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Citations

Mar 27, 1999·Oral Microbiology and Immunology·A MombelliE F Corbet
Jul 11, 2006·Journal of Periodontal Research·C PadillaC Descouvieres
Aug 17, 1999·Microbiology and Immunology·M SakamotoT Nakase
Oct 8, 1999·Journal of Dental Research·B EhmkeT F Flemmig
Aug 7, 2009·Journal of Periodontology·Fernando VerdugoJosé Pontón
Dec 22, 1999·Journal of Clinical Periodontology·B EhmkeT F Flemmig

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