Browse > Article

Development of prevotella intermedia ATCC 49046 Strain-Specific PCR Primer Based on a Pig6 DNA Probe  

Jeong Seung-U (Department of Periodontology, College of Dentistry, Chosun University)
Yoo So-Young (Department of Oral Biochemistry, College of Dentistry, Chosun University)
Kang Sook-Jin (Department of Oral Biochemistry, College of Dentistry, Chosun University)
Kim Mi-Kwang (Department of Oral Biochemistry, College of Dentistry, Chosun University)
Jang Hyun-Seon (Department of Periodontology, College of Dentistry, Chosun University)
Lee Kwang-Yong (Undergraduate student, College of Dentistry, Chosun University)
Kim Byung-Ok (Department of Periodontology, College of Dentistry, Chosun University)
Kook Joong-Ki (Department of Oral Biochemistry, College of Dentistry, Chosun University)
Publication Information
Korean Journal of Microbiology / v.42, no.2, 2006 , pp. 89-94 More about this Journal
Abstract
The purpose of this study is to develop the strain-specific PCR primers for the identification of prevotella inter-media ATCC 49046 which is frequently used in the pathogenesis studies of periodontitis. The Hind III-digested genomic DNA of P. intermedia ATCC 49046 were cloned by random cloning method. The specificity of cloned DNA fragments were determined by Southern blot analysis. The nucleotide sequence of cloned DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pig6 DNA probe were hybridized with the genomic DNA from P. intermedia strains (ATCC $25611^T$ and 49046) isolated from the Westerns, not the strains isolated from Koreans. The Pig6 DNA probe were consisted of 813 bp. Pig6-F3 and Pig6-R3 primers, designed base on the nucleotide Sequences Of Pig6 DNA Probe, were 3150 specific to the only both P. intermedia ATCC $25611^T$ and P. intermedia ATCC 49046. In the other hand, Pig6-60F and Pig6-770R primers were specific to the only P. intermedia ATCC 49046. The two PCR primer sets could detect as little as 4 pg of chromosomal DNA of P. intermedia. These results indicate that Pig6-60F and Pig6-770R primers have proven useful for the identification of P. intermedia ATCC 49046, especially with regard to the maintenance of the strain.
Keywords
detection; DNA probe Pig6; PCR primer; Prevotella intermedia ATCC 49046;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
연도 인용수 순위
1 Kook, J.K., T. akamoto, K. Nishi, M.K. Kim, J.H. Seong, Y.N. Son, and D.K. Kim. 2005. Detection of Tannerella forsythia and/ or Prevotella intermedia might be useful for microbial predictive markers for the outcome of initial periodontal treatment in Koreans. Microbiol. Immunol. 49, 9-16. Erratum in: Microbiol. Immunol. 2005. 49, 295   DOI
2 Kook, J.K., J.J. Han, H.S. Kim, J.H. Seong, D.K. Kim, D.H. Baek, and S.J. Choe. 2003. Cloning of a potentially strain-specific DNA probe of Prevotella intermedia ATCC 25611 by inverted dot blot hybridization screening method. J. Microbiol. Biotechnol. 13, 282-286
3 Krieg, N.R. 2001. Identification of Procaryotes, p. 33-38. In G. Garrity (ed.), Bergey's Manual of Systematic Bacteriology, 2th ed. Springer Verlag, New York, U.S.A
4 Slots, J. and M.A. Listgarten. 1988. Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases. J. Clin. Periodontol. 15, 85- 93   DOI
5 Dzink, J.L., S.S. Socransky, and A.D. Haffajee. 1988. The predominant cultivable microbiota of active and inactive lesions of destructive periodontal diseases. J. Clin. Periodontol. 15, 316-323   DOI
6 Shin, Y.K., S.U. Jeong, S.Y. Yoo, M.K. Kim, H.S. Kim, B.O. Kim, D.K. Kim, H.K. Hwang, and J.K. Kook. 2004. Pi30 DNA probe may be useful for the identification of Prevotella intermedia at the species or strain level. Microbiol. Immunol. 48, 931-936   DOI
7 Socransky, S.S., A.D. Haffajee, J.L. Dzink, and J.D. Hillman. 1988. Associations between microbial species in subgingival plaque samples. Oral Microbiol. Immunol. 3, 1-7   DOI   ScienceOn
8 Kim, H.S., S.K. Song, S.Y. Yoo, D.C. Jin, H.S. Shin, C.K. Lim, M.S. Kim, J.S. Kim, S.J. Choe, and J.K. Kook. 2005. Develop-ment of strain-specific PCR primers based on a DNA probe Fu12 for the identification of Fusobacterium nucleatum subsp. nuclea-tum ATCC $25586^{T}$. J. Microbiol. 43, 331-336. Erratum in: J. Microbiol. 2005. 43, 473   과학기술학회마을
9 Papapanou, P.N., A.M. Neiderud, A. Papadimitriou, J. Sandros, and G. Dahlen. 2000. 'Checkerboard' assessments of periodontal microbiota and serum antibody responses: a case-control study. J. Periodontol. 71, 885-897   DOI   ScienceOn
10 Milsom, S.E., S.V. Sprague, D. Dymock, A.J. Weightman, and W.G. Wade. 1996. Rapid differentiation of Prevotella intermedia and P. nigrescens by 16S rDNA PCR-RFLP. J. Med. Microbiol. 44, 41-43   DOI
11 Okamoto, M., N. Maeda, K. Kondo, and K.P. Leung. 1999. Hemolytic and hemagglutinating activities of Prevotella intermedia and Prevotella nigrescens. FEMS Microbiol. Lett. 178, 299- 304   DOI
12 Slots, J., L. Bragd, M. Wikstrom, and G. Dahlen. 1986. The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in destructive periodontal disease in adults. J. Clin. Periodontol. 13, 570-577   DOI
13 Kim, S.J., M.S. Ha, E.Y. Choi, J.I. Choi, and I.S. Choi. 2004. Prevotella intermedia lipopolysaccharide stimulates release of nitric oxide by inducing expression of inducible nitric oxide synthase. J. Periodontal Res. 39, 424-431   DOI   ScienceOn
14 Saito, K., N. Takahashi, H. Horiuchi, and T. Yamada. 2001. Effects of glucose on formation of cytotoxic end-products and proteolytic activity of Prevotella intermedia, Prevotella nigrescens and Porphyromonas gingivalis. J. Periodontal Res. 36, 355-360   DOI   ScienceOn
15 Paster, B.J., S.K. Boches, J.L. Galvin, R.E. Ericson, C.N. Lau, V.A. Levanos, A. Sahasrabudhe, and F.E. Dewhirst. 2001. Bacterial diversity in human subgingival plaque. J. Bacteriol. 183, 3770- 3783   DOI   ScienceOn
16 Dorn, B.R., K.L. Leung, and A. Progulske-Fox. 1998. Invasion of human oral epithelial cells by Prevotella intermedia. Infect. Immun. 66, 6054-6057