Similar articles

Abstract: Objective: The aim of this study was to investigate subgingival infection frequencies of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans strains with genetic variation in Chinese chronic periodontitis (CP) patients and to evaluate its correlation with clinical parameters. Methods: Two multiplex polymerase chain reaction (PCR) assays were developed to detect the 16SrDNA, collagenase (prtC) and fimbria (fimA) genes of P. gingivalis and the 16SrDNA, leukotoxin (lktA) and fimbria-associated protein (fap) genes of A. actinomycetemcomitans in 60 sulcus samples from 30 periodontal healthy subjects and in 122 subgingival plaque samples from 61 patients with CP. The PCR products were further T-A cloned and sent for nucleotide sequence analysis. Results: The 16SrDNA, prtC and fimA genes of P. gingivalis were detected in 92.6%, 85.2% and 80.3% of the subgingival plaque samples respectively, while the 16SrDNA, lktA and fap genes of A. actinomycetemcomitans were in 84.4%, 75.4% and 50.0% respectively. Nucleotide sequence analysis showed 98.62%~100% homology of the PCR products in these genes with the reported sequences. P. gingivalis strains with prtC+/fimA+ and A. actinomycetemcomitans with lktA+ were predominant in deep pockets (>6 mm) or in sites with attachment loss ≥5 mm than in shallow pockets (3~4 mm) or in sites with attachment loss ≤2 mm (P<0.05). P. gingivalis strains with prtC+/fimA+ also showed higher frequency in gingival index (GI)=3 than in GI=1 group (P<0.05). Conclusion: Infection of P. gingivalis with prtC+/fimA+ and A. actinomycetemcomitans with lktA+ correlates with periodontal destruction of CP in Chinese. Nonetheless P. gingivalis fimA, prtC genes and A. actinomycetemcomitans lktA gene are closely associated with periodontal destruction, while A. actinomycetemcomitans fap gene is not.

[1] Amano, A., 2003. Molecular interaction of Porphyromonas gingivalis with host cells: implication for the microbial pathogenesis of periodontal disease. J. Periodontol., 74(1):90-96.

[2] Amano, A., Nakagawa, I., Kataoka, K., Morisaki, I., Hamada, S., 1999. Distribution of Porphyromonas gingivalis strains with fimA genotypes in periodontitis patients. J. Clin. Microbiol., 37(5):1426-1430.

[3] Amano, A., Kuboniwa, M., Nakagawa, I., Akiyama, S., Morisaki, I., Hamada, S., 2000. Prevalence of specific genotypes of Porphyromonas gingivalis fimA and periodontal health status. J. Dent. Res., 79(9):1664-1668.

[4] Amano, A., Nakagawa, I., Okahashi, N., Hamada, N., 2004. Variations of Porphyromonas gingivalis fimbriae in relation to microbial pathogenesis. J. Periodontal Res., 39(2):136-142.

[5] Armitage, G.C., 1999. Development of a classification system for periodontal disease and conditions. Ann. Periodontol., 4(1):1-6.

[6] Beikler, T., Peters, U., Prajaneh, S., Prior, K., Ehmke, B., Flemmig, T.F., 2003a. Prevalence of Porphyromonas gingivalis fimA genotypes in Caucasians. Eur. J. Oral Sci., 111(5):390-394.

[7] Beikler, T., Ehmke, B., Wittstock, M., Schmidt, H., Karch, H., Flemmig, T.F., 2003b. Serum antibody reactivity against recombinant PrtC of Porphyromonas gingivalis following periodontal therapy. J. Periodontal Res., 38(3):276-281.

[8] Bodinka, A., Schmidt, H., Henkel, B., Flemmig, T.F., Klaiber, B., Karch, H., 1994. Polymerase chain reaction for the identification of Porphyromonas gingivalis collagenase genes. Oral Microbiol. Immunol., 9(3):161-165.

[9] Chen, L.L., Wu, Y.M., Yan, J., Sun, W.L., Sun, Y.Z., Ojcius, D., 2005. Association between coinfection of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Treponema denticola and periodontal tissue destruction in chronic periodontitis. Chin. Med. J. (Engl.), 118(11):915-921.

[10] Cortelli, J.R., Cortelli, S.C., Jordan, S., Haraszthy, V.I., Zambon, J.J., 2005a. Prevalence of periodontal pathogens in Brazilians with aggressive or chronic periodontitis. J. Clin. Periodontol., 32(8):860-866.

[11] Cortelli, S.C., Feres, M., Rodrigues, A.A., Aquino, D.R., Shibli, J.A., Cortelli, J.R., 2005b. Detection of Actinobacillus actinomycetemcomitans in unstimulated saliva of patients with chronic periodontitis. J. Periodontol., 76(2):204-209.

[12] Darout, I.A., Skaug, N., Albandar, J.M., 2003. Subgingival microbiota levels and their associations with periodontal status at the sampled sites in an adult Sudanese population using miswak or toothbrush regularly. Acta Odontol. Scand., 61(2):115-122.

[13] Fujiwara, T., Nakagawa, I., Morishima, S., Takahashi, I., Hamada, S., 1994. Inconsistency between the fimbrillin gene and the antigenicity of lipopolysaccharides in selected strains of Porphyromonas gingivalis. FEMS Microbiol. Lett., 124(3):333-341.

[14] Gafan, G.P., Lucas, V.S., Roberts, G.J., Petrie, A., Wilson, M., Spratt, D.A., 2004. Prevalence of periodontal pathogens in dental plaque of children. J. Clin. Microbiol., 42(9):4141-4146.

[15] Guthmiller, J.M., Lally, E.T., Korostoff, J., 2001. Beyond the specific plaque hypothesis: are highly leukotoxic strains of Actinobacillus actinomycetemcomitans a paradigm for periodontal pathogenesis? Crit. Rev. Oral Biol. Med., 12(2):116-124.

[16] Hamlet, S.M., Cullinan, M.P., Westerman, B., Lindeman, M., Bird, P.S., Palmer, J., Seymour, G.J., 2001. Distribution of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in an Australian population. J. Clin. Periodontol., 28(12):1163-1171.

[17] Imamura, T., Travis, J., Potempa, J., 2003. The biphasic virulence activities of gingipains: activation and inactivation of host proteins. Curr. Protein Pept. Sci., 4(6):443-450.

[18] Ishihara, K., Honma, K., Miura, T., 1997. Cloning and sequence analysis of the fimbriae associated protein (fap) gene from Actinobacillus actinomycetemcomitans. Microb. Pathog., 23(2):63-69.

[19] Johansson, A., Buhlin, K., Koski, R., Gustafsson, A., 2005. The immunoreactivity of systemic antibodies to Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in adult periodontitis. Eur. J. Oral Sci., 113(3):197-202.

[20] Kato, T., Takahashi, N., Kuramitsu, H.K., 1992. Sequence analysis and characterization of the Porphyromonas gingivalis prtC gene, which expresses a novel collagenase activity. J. Bacteriol., 174(12):3889-3895.

[21] Lakio, L., Kuula, H., Dogan, B., Asikainen, S., 2002. Actinobacillus actinomycetemcomitans proportion of subgingival bacterial flora in relation to its clonal type. Eur. J. Oral Sci., 110(3):212-217.

[22] Leung, W.K., Ngai, V.K., Yau, J.Y., Cheung, B.P., Tsang, P.W., Corbet, E.F., 2005. Characterization of Actinobacillus actinomycetemcomitans isolated from young Chinese aggressive periodontitis patients. J. Periodontal Res., 40(3):258-268.

[23] Liu, L., Wen, X., He, H., Shi, J., Ji, C., 2003. Species-specific DNA probe for the detection of Porphyromonas gingivalis from adult Chinese periodontal patients and healthy subjects. J. Periodontol., 74(7):1000-1006.

[24] Nogueira Moreira, A., Chiappe, V., Fernandez Caniggia, L., Alonso, C., Piovano, S., 2004. Clinical and microbiological associations in chronic periodontitis. Acta Odontol. Latinoam., 17(1-2):15-21.

[25] Noiri, Y., Li, L., Ebisu, S., 2001. The localization of periodontal-disease-associated bacteria in human periodontal pockets. J. Dent. Res., 80(10):1930-1934.

[26] Nonnenmacher, C., Mutters, R., de Jacoby, L.F., 2001. Microbiological characteristics of subgingival microbiota in adult periodontitis, localized juvenile periodontitis and rapidly progressive periodontitis subjects. Clin. Microbiol. Infect., 7(4):213-217.

[27] Potempa, J., Banbula, A., Travis, J., 2000. Role of bacterial proteinases in matrix destruction and modulation of host responses. Periodontology 2000, 24(1):153-192.

[28] Poulsen, K., Ennibi, O.K., Haubek, D., 2003. Improved PCR for detection of the highly leukotoxic JP2 clone of Actinobacillus actinomycetemcomitans in subgingival plaque samples. J. Clin. Microbiol., 41(10):4829-4832.

[29] Slots, J., Ashimoto, A., Flynn, M.J., Li, G.L., Chen, C., 1995. Detection of putative pathogens in subgingival specimens by 16S ribosomal DNA amplification with the polymerase chain reaction. Clin. Infect. Dis., 20(Suppl. 2):304-307.

[30] Takeuchi, Y., Umeda, M., Sakamoto, M., Benno, Y., Huang, Y., Ishikawa, I., 2001. Treponema socranskii, Treponema denticola, and Porphyromonas gingivalis are associated with severity of periodontal tissue destruction. J. Periodontol., 72(10):1354-1363.

[31] Tamura, K., Nakano, K., Nomura, R., Miyake, S., Nakagawa, I., Amano, A., Ooshima, T., 2005. Distribution of Porphyromonas gingivalis fimA genotypes in Japanese children and adolescents. J. Periodontol., 76(5):674-679.

[32] Tan, K.S., Woo, C.H., Ong, G., Song, K.P., 2001. Prevalence of Actinobacillus actinomycetemcomitans in an ethnic adult Chinese population. J. Clin. Periodontol., 28(9):886-890.

[33] van der Ploeg, J.R., Giertsen, E., Ludin, B., Morgeli, C., Zinkernagel, A.S., Gmur, R., 2004. Quantitative detection of Porphyromonas gingivalis fimA genotypes in dental plaque. FEMS Microbiol. Lett., 232(1):31-37.

[34] Watanabe, K., Frommel, T.O., 1993. Detection of Porphyromonas gingivalis in oral plaque samples by use of the polymerase chain reaction. J. Dent. Res., 72(6):1040-1044.

[35] Watanabe, K., Frommel, T.O., 1996. Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Treponema denticola detection in oral plasque samples using the polymerase chain reaction. J. Clin. Periodontol., 23(3):212-219.

[36] Wittstock, M., Schmidt, H., Flemmig, T.F., Karch, H., 2000. Heterogeneity of the prtC gene of Porphyromonas gingivalis. Oral Microbiol. Immunol., 15(1):33-39.

[37] Yang, H.W., Huang, Y.F., Chan, Y., Chou, M.Y., 2005. Relationship of Actinobacillus actinomycetemcomitans serotypes to periodontal condition: prevalence and proportions in subgingival plaque. Eur. J. Oral Sci., 113(1):28-33.

[38] Yuan, K., Chang, C.J., Hsu, P.C., Sun, H.S., Tseng, C.C., Wang, J.R., 2001. Detection of putative periodontal pathogens in non-insulin-dependent diabetes mellitus and non-diabetes mellitus by polymerase chain reaction. J. Periodontal Res., 36(1):18-24.