Browse > Article

Identification of Actinobacillus actinomycetemcomitans Using Species-Specific 16S rDNA Primers  

Kim Su Gwan (Department of Oral and Maxillofacial Surgery, College of Dentistry, Chosun University)
Kim Soo Heung (Department of Oral and Maxillofacial Surgery, College of Dentistry, Chosun University)
Kim Mi Kwang (Department of Oral Biochemistry, College of Dentistry, Chosun University)
Kim Hwa Sook (Department of Oral Biochemistry, College of Dentistry, Chosun University)
Kook Joong Ki (Department of Oral Biochemistry, College of Dentistry, Chosun University)
Publication Information
Journal of Microbiology / v.43, no.2, 2005 , pp. 209-212 More about this Journal
Abstract
The purpose of this study was to develop species-specific PCR primers for use in the identification and detection of Actinobacillus actinomycetemcomitans. These primers target variable regions of the 168 ribosomal RNA coding gene (rDNA). We assessed the specificity of the primers against 9 A. actinomycetemcomitans strains and 11 strains (3 species) of the Haemophilus genus. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC$ 33384^$T Our obtained data revealed that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 4 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these PCR primers are incredibly sensitive, and should prove suitable for application in epidemiological studies, as well as the diagnosis and monitoring of periodontal pathogens after treatment for periodontitis.
Keywords
Citations & Related Records

Times Cited By Web Of Science : 6  (Related Records In Web of Science)
Times Cited By SCOPUS : 6
연도 인용수 순위
1 Conrads, G., T.F. Flemmig, I. Seyfarth, F. Lampert, and R. Lutticken. 1999. Simultaneous detection of Bacteroides forsythus and Prevotella intermedia by 16S rRNA gene-directed multiplex PCR. J. Clin. Microbiol. 37, 1621-1624   PUBMED
2 Lally, E.T., I.R. Kieba, D.R. Demuth, J. Rosenbloom, E.E. Golub, N.S. Taichman, and C.W. Gibson. 1989. Identification and expression of the Actinobacillus actinomycetemcomitans leukotoxin gene. Biochem. Biophys. Res. Commun. 159, 256-262   DOI   ScienceOn
3 Seong, J.H., D.K. Kim, K.Y. Shin, B.O. Kim, M.S. Kim, M.K. Kim, and J.K. Kook. 2003. Simultaneous detection of Prevotella intermedia and Prevotella nigrescens by 16S rDNA-directed multiplex PCR. J. Korean Acad. Dent. Health 26, 523-533
4 Tran, S.D. and J.D. Rudney. 1999. Improved multiplex PCR using conserved and species-specific 16S rRNA gene primers for simultaneous detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis. J. Clin. Microbiol. 37, 3504-3508   PUBMED
5 Asikainen, S. 1986. Occurrence of Actinobacillus actinomycetemcomitans and spirochetes in relation to age in localized juvenile periodontitis. J. Periodontol. 57, 537-541   DOI   PUBMED
6 Kiley, P. and S.C. Holt. 1980. Characterization of the lipopolysaccharide from Actinobacillus actinomycetemcomitans Y4 and N27. Infect. Immun. 30, 862-873   PUBMED
7 Kook, J.-K., M.-K. Kim, J.-H. Seong, D.-K. Kim, B.-O. Kim, J.-C. Park, K.-K. Kim, S.-J. Choe, and B.-M. Min. 2003. A new method for rapid screening of bacterial species- or subspeciesspecific DNA probes. FEMS Microbiol. Lett. 219, 121-127   DOI   ScienceOn
8 Zambon, J.J. 1985. Actinobacillus actinomycetemcomitans in human periodontal disease. J. Clin. Periodontol. 12, 1-20   DOI   PUBMED
9 Robertson, P.B., M. Lantz, P.T. Marucha, K.S. Kornman, C.L. Trummel, and S.C. Holt. 1982. Collagenolytic activity associated with Bacteroides species and Actinobacillus actinomycetemcomitans. J. Periodontal Res. 17, 275-283   DOI   PUBMED
10 Darveau, R.P., A. Tanner, and R.C. Page. 1997. The microbial challenge in periodontitis. Periodontol. 2000 14, 12-32   DOI   PUBMED   ScienceOn
11 Woese, C.R. 1987. Bacterial evolution. Microbiol. Rev. 51, 221-271   PUBMED
12 Ashimoto, A., C. Chen, I. Bakker, and J. Slots. 1996. Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions. Oral Microbiol. Immunol. 11, 266-273   DOI   ScienceOn
13 Tonjum, T., and R. Haas. 1993. Identification of Actinobacillus actinomycetemcomitans by leukotoxin gene-specific hybridization and polymerase chain reaction assays. J. Clin. Microbiol. 31, 1856-1859   PUBMED
14 Lippke, J.A., M.N. Strzempko, F.F. Raia, S.L. Simon, and C.K. French. 1987. Isolation of intact high-molecular-weight DNA by using guanidine isothiocyanate. Appl. Environ. Microbiol. 53, 2588-2589   PUBMED
15 Ashkenazi, M., R.R. White, and D.K. Dennison.1992. Neutrophil modulation by Actinobacillus actinomycetemcomitans. I. Chemotaxis, surface receptor expression and F-actin polymerization. J. Periodontal Res. 27, 264-273   DOI   PUBMED
16 Slots, J., A. Ashimoto, M.J. Flynn, G. Li, and C. Chen. 1995. Detection of putative periodontal pathogens in subgingival specimens by 16S ribosomal DNA amplification with the polymerase chain reaction. Clin. Infect. Dis. 20(Suppl 2), S304-307   DOI
17 Tran, S.D. and J.D. Rudney. 1996. Multiplex PCR using conserved and species-specific 16S rRNA gene primers for simultaneous detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. J. Clin. Microbiol. 34, 2674-2678   PUBMED
18 van Winkelhoff, A.J. and J. Slots. 1999. Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in nonoral infections. Periodontol. 2000 20, 122-135   DOI   PUBMED   ScienceOn
19 Mandell, R.L. 1984. A longitudinal microbiological investigation of Actinobacillus actinomycetemcomitans and Eikenella corrodens in juvenile periodontitis. Infect. Immun. 45, 778-780   PUBMED
20 Sugai, M., T. Kawamoto, S.Y. Peres, Y. Ueno, H. Komatsuzawa, T. Fujiwara, H. Kurihara, H. Suginaka, and E. Oswald. 1998. The cell cycle-specific growth-inhibitory factor produced by Actinobacillus actinomycetemcomitans is a cytolethal distending toxin. Infect. Immun. 66, 5008-5019
21 Genco, R.J., J.J. Zambon, and P.A. Murray. 1985. Serum and gingival fluid antibodies as adjuncts in the diagnosis of Actinobacillus actinomycetemcomitans-associated periodontal disease. J. Periodontol. 56, 1-50   PUBMED