Subgingival periodontal pathogens associated with chronic periodontitis in Yemenis

BMC Oral Health

Ibrahim Ghandour²  Ali K Al-Sharabi³  Hussein M Shuga-Aldin³  Nezar N Al-hebshi¹ 

[1] Molecular Research Laboratory, Faculty of Medical Sciences, University of Science and Technology, Sana’a, Yemen;Department of Periodontology, Faculty of Dentistry, Khartoum University, Khartoum, Sudan;Department of Periodontology, Oral Pathology, Oral Medicine and Radiology; Faculty of Dentistry, University of Sana’a, Sana’a, Yemen

关键词: Periodontitis;    Parvimonas micra;    Real-time PCR;    Pathogens;    Microbiology;   

Others  :  1121459
DOI  :  10.1186/1472-6831-14-13

 received in 2013-11-12, accepted in 2014-02-14,  发布年份 2014

【 摘 要 】

Background

Subgingival microbial profile associated with periodontitis has been reported to significantly differ by geographical location. The purpose of this study was to assess the association between a panel of putative periodontal bacterial pathogens and chronic periodontitis among Yemenis.

Methods

Subgingival DNA samples were obtained from diseased and healthy sites of 20 non-smoking, moderate to severe chronic periodontitis subjects. Absolute counts (bacterial DNA copies per sample) and relative counts (% total bacteria) of seven periopathogenic species/genera representative of the red and orange complexes were determined using Taqman q-PCR assays.

Results

The q-PCR assays showed excellent linearity (R² > 0.99) and a sensitivity of 100 copies/sample. The detection rate was 100% for all tested species/genera except for P. gingivalis and A. actinomycetemcomitans that were detected at 97.5% and 67.5%, respectively. The median log absolute counts were in the range of 2.41-6.53 copies per sample while median relative counts were in the range of 0.001-0.77%, both being highest for fusobacteria and lowest for A. actinomycetemcomitans. Significant interspecies correlations were observed. Adjusting for multiple comparisons (P≤0.0063), only T. forsythia, T. denticola and P. micra maintained significant association with periodontal destruction. The latter species, however, showed the strongest association and was found in higher proportions at the periodontitis sites across all subjects (3.39 median fold increase). No significant differences were observed for P. gingivalis.

Conclusions

P. micra rather than P. gingivalis appears as a keystone pathogen in this Yemeni Sample. However, these findings need to be validated in a larger-scale study before they can be claimed to represent ethnic variations in pathogens’ association with periodontitis.

【 授权许可】

   
2014 Al-hebshi et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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【 参考文献 】

• [1]Darveau RP: Periodontitis: a polymicrobial disruption of host homeostasis. Nat Rev Microbiol 2010, 8(7):481-490.
• [2]Moore WE, Moore LV: The bacteria of periodontal diseases. Periodontol 2000 1994, 5:66-77.
• [3]Kumar PS, Griffen AL, Moeschberger ML, Leys EJ: Identification of candidate periodontal pathogens and beneficial species by quantitative 16S clonal analysis. J Clin Microbiol 2005, 43(8):3944-3955.
• [4]Paster BJ, Boches SK, Galvin JL, Ericson RE, Lau CN, Levanos VA, Sahasrabudhe A, Dewhirst FE: Bacterial diversity in human subgingival plaque. J Bacteriol 2001, 183(12):3770-3783.
• [5]Dewhirst FE, Chen T, Izard J, Paster BJ, Tanner AC, Yu WH, Lakshmanan A, Wade WG: The human oral microbiome. J Bacteriol 2010, 192(19):5002-5017.
• [6]Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr: Microbial complexes in subgingival plaque. J Clin Periodontol 1998, 25(2):134-144.
• [7]Dewhirst FE, Tamer MA, Ericson RE, Lau CN, Levanos VA, Boches SK, Galvin JL, Paster BJ: The diversity of periodontal spirochetes by 16S rRNA analysis. Oral Microbiol Immunol 2000, 15(3):196-202.
• [8]Rylev M, Kilian M: Prevalence and distribution of principal periodontal pathogens worldwide. J Clin Periodontol 2008, 35(8 Suppl):346-361.
• [9]Mineoka T, Awano S, Rikimaru T, Kurata H, Yoshida A, Ansai T, Takehara T: Site-specific development of periodontal disease is associated with increased levels of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in subgingival plaque. J Periodontol 2008, 79(4):670-676.
• [10]Yoshida A, Kawada M, Suzuki N, Nakano Y, Oho T, Saito T, Yamashita Y: TaqMan real-time polymerase chain reaction assay for the correlation of Treponema denticola numbers with the severity of periodontal disease. Oral Microbiol Immunol 2004, 19(3):196-200.
• [11]Lyons SR, Griffen AL, Leys EJ: Quantitative real-time PCR for Porphyromonas gingivalis and total bacteria. J Clin Microbiol 2000, 38(6):2362-2365.
• [12]Kuboniwa M, Amano A, Kimura KR, Sekine S, Kato S, Yamamoto Y, Okahashi N, Iida T, Shizukuishi S: Quantitative detection of periodontal pathogens using real-time polymerase chain reaction with TaqMan probes. Oral Microbiol Immunol 2004, 19(3):168-176.
• [13]Haffajee AD, Bogren A, Hasturk H, Feres M, Lopez NJ, Socransky SS: Subgingival microbiota of chronic periodontitis subjects from different geographic locations. J Clin Periodontol 2004, 31(11):996-1002.
• [14]World Health Organization: Oral health surveys: basic methods. 4th edition. Geneva: World Health Organization; 1997.
• [15]Silness J, Loe H: Periodontal disease in pregnancy. Ii. Correlation between oral hygiene and periodontal condtion. Acta Odontol Scand 1964, 22:121-135.
• [16]Holland PM, Abramson RD, Watson R, Gelfand DH: Detection of specific polymerase chain reaction product by utilizing the 5′––3′ exonuclease activity of Thermus aquaticus DNA polymerase. Proc Natl Acad Sci U S A 1991, 88(16):7276-7280.
• [17]Al-Hebshi NN, Al-Sharabi AK, Shuga-Aldin HM, Al-Haroni M, Ghandour I: Effect of khat chewing on periodontal pathogens in subgingival biofilm from chronic periodontitis patients. J Ethnopharmacol 2010, 132(3):564-569.
• [18]Suzuki N, Yoshida A, Saito T, Kawada M, Nakano Y: Quantitative microbiological study of subgingival plaque by real-time PCR shows correlation between levels of Tannerella forsythensis and Fusobacterium spp. J Clin Microbiol 2004, 42(5):2255-2257.
• [19]Martin FE, Nadkarni MA, Jacques NA, Hunter N: Quantitative microbiological study of human carious dentine by culture and real-time PCR: association of anaerobes with histopathological changes in chronic pulpitis. J Clin Microbiol 2002, 40(5):1698-1704.
• [20]Al-Otaibi M, Al-Harthy M, Gustafsson A, Johansson A, Claesson R, Angmar-Mansson B: Subgingival plaque microbiota in Saudi Arabians after use of miswak chewing stick and toothbrush. J Clin Periodontol 2004, 31(12):1048-1053.
• [21]Darout IA, Skaug N, Albandar JM: 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 2003, 61(2):115-122.
• [22]Nonnenmacher C, Dalpke A, Mutters R, Heeg K: Quantitative detection of periodontopathogens by real-time PCR. J Microbiol Methods 2004, 59(1):117-125.
• [23]Lin CH, Chen YC, Pan TM: Quantification bias caused by plasmid DNA conformation in quantitative real-time PCR assay. PLoS One 2011, 6(12):e29101.
• [24]Ximenez-Fyvie LA, Haffajee AD, Socransky SS: Microbial composition of supra- and subgingival plaque in subjects with adult periodontitis. J Clin Periodontol 2000, 27(10):722-732.
• [25]Moore WE, Holdeman LV, Smibert RM, Hash DE, Burmeister JA, Ranney RR: Bacteriology of severe periodontitis in young adult humans. Infect Immun 1982, 38(3):1137-1148.
• [26]Hajishengallis G, Lamont RJ: Beyond the red complex and into more complexity: the polymicrobial synergy and dysbiosis (PSD) model of periodontal disease etiology. Mol Oral Microbiol 2012, 27(6):409-419.
• [27]Hajishengallis G, Liang S, Payne MA, Hashim A, Jotwani R, Eskan MA, McIntosh ML, Alsam A, Kirkwood KL, Lambris JD, et al.: Low-abundance biofilm species orchestrates inflammatory periodontal disease through the commensal microbiota and complement. Cell Host Microbe 2011, 10(5):497-506.
• [28]Hajishengallis G, Darveau RP, Curtis MA: The keystone-pathogen hypothesis. Nat Rev Microbiol 2012, 10(10):717-725.
• [29]Colombo AP, Boches SK, Cotton SL, Goodson JM, Kent R, Haffajee AD, Socransky SS, Hasturk H, Van Dyke TE, Dewhirst F, et al.: Comparisons of subgingival microbial profiles of refractory periodontitis, severe periodontitis, and periodontal health using the human oral microbe identification microarray. J Periodontol 2009, 80(9):1421-1432.
• [30]Kumar PS, Leys EJ, Bryk JM, Martinez FJ, Moeschberger ML, Griffen AL: Changes in periodontal health status are associated with bacterial community shifts as assessed by quantitative 16S cloning and sequencing. J Clin Microbiol 2006, 44(10):3665-3673.
• [31]Rams TE, Feik D, Listgarten MA, Slots J: Peptostreptococcus micros in human periodontitis. Oral Microbiol Immunol 1992, 7(1):1-6.
• [32]Belstrom D, Fiehn NE, Nielsen CH, Kirkby N, Twetman S, Klepac-Ceraj V, Paster BJ, Holmstrup P: Differences in bacterial saliva profile between periodontitis patients and a control cohort. J Clin Periodontol 2014, 41(2):104-112.