Prevalence and molecular diversity of <i>Archaea</i> in subgingival pockets of periodontitis patients

Li, C. L.; Liu, D. L.; Jiang, Yijie; Zhou, Yan; Zhang, M. Z.; Jiang, Wenxin; Liu, B.; Ji, Liang

doi:10.1111/j.1399-302x.2009.00514.x

Cited by 74 publications

(85 citation statements)

References 24 publications

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“…Total bacterial control indicated that for a C T value of Ͼ35.5, M. oralis detection was not interpretable. Based on these controls, the detection of M. oralis DNA in healthy individuals (C T Յ 35.5) was possible, in contrast to findings of previous studies (12,13,17,18), and a significant correlation was observed between the M. oralis load and periodontitis severity.…”

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confidence: 52%

“…PCR systems previously used to detect methanogens in the subgingival plaque (6,8,12,13) were not completely specific for M. oralis. Indeed, other methanogens, including Methanobrevibacter smithii (14), Methanosphaera stadtmanae (6), Methanobacterium curvum/Methanobacterium congolense, and Methano-sarcina mazeii (15,16), have been detected in this situation.…”

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confidence: 99%

“…The M. oralis cell number correlated significantly with the periodontal clinical score, with an r value of 0.527 (n ϭ 30) and a P value of 0.003. The significance, analysis of variance (ANOVA) test, or the Kruskal-Wallis test (2-tailed) correlation was 0.003.PCR systems previously used to detect methanogens in the subgingival plaque (6,8,12,13) were not completely specific for M. oralis. Indeed, other methanogens, including Methanobrevibacter smithii (14), Methanosphaera stadtmanae (6), Methanobacterium curvum/Methanobacterium congolense, and Methano-…”

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Real-Time PCR Quantification of Methanobrevibacter oralis in Periodontitis

et al. 2013

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bA real-time PCR assay developed to quantify Methanobrevibacter oralis indicated that its inoculum significantly correlated with periodontitis severity (P ‫؍‬ 0.003), despite a nonsignificant difference in prevalence between controls (3/10) and patients (12/22) (P ‫؍‬ 0.2, Fisher test). The M. oralis load can be used as a biomarker for periodontitis. Periodontitis is an anaerobic infection possibly leading to loss of teeth (1-3). It likely results from infection with microbial complexes comprising methanogens (4, 5), mostly Methanobrevibacter oralis (6)(7)(8). In this study, we correlated the M. oralis load as measured by real-time PCR in subgingival plaque with the severity of periodontitis.All patients diagnosed with periodontitis from October 2011 to June 2012 were included. The control individuals with generally healthy gingiva were volunteers. All patients underwent interviews for medical and dental history, intraoral examination to determine bleeding on probing (BOP), probing depth (PD), and plaque index (PI), and radiographs (9); periodontal status was also scored as described previously (10) ( Table 1). All individuals signed an informed-consent form, and the ethics committee of the IFR 48, University of Aix-Marseille, approved the protocol.Subgingival plaque specimens (50 l) collected from 3-to 12-mm periodontal pockets in both patients and controls were suspended in 1 ml Tris-HCl (0.05 M, pH 7.5) buffer. After homogenization, a 250-l aliquot was shaken with 0.3 g of acid-washed beads (Յ106 mm; Sigma, Saint-Quentin-Fallavier, France) in a FastPrep-24 instrument (MP Biomedical Europe, Illkirch, France) at a speed of 6.5 m/s (full speed) for 90 s. The supernatant was incubated overnight at 56°C with 180 ml of lysis buffer and 25 l proteinase K (20 mg/ml) in the Qiagen EZ1 DNA tissue kit (Qiagen, Courtaboeuf, France). After a second cycle of mechanical lysis, the supernatant was incubated for 10 min at 100°C, and total DNA was then extracted using the same kit. Negative controls consisting of sterile DNA-free water were introduced in all the manipulations. The specificity of the M. oralis-cnp602P probe (6-carboxyfluorescein [FAM]-5=AGCAGTGCACCTGCTGATA TGGAAGG-3=) (Applied Biosystems, Courtaboeuf, France) and the primer pair M. oralis-cnp602F (5=-GCTGGTGTAATCGAAC CTAAACG-3=) and M. oralis-cnp602R (5=-CACCCATACCCGG ATCCATA-3=) (Eurogentec, Seraing, Belgium) was verified in silico using the BLAST program at NCBI (http://www.ncbi.nlm .nih.gov/BLAST) and further experimentally ensured by incorporating DNA extracted from Methanobrevibacter smithii ATCC 35061 T , Methanosphaera stadtmanae ATCC 43021 T , Methanomassiliicoccus luminyensis CSURP135 T , Escherichia coli, Salmonella enterica, Staphylococcus aureus, and Treponema denticola ATCC 35405T . Real-time PCR assays were performed with a CFX96 Touch real-time PCR detection system (Bio-Rad, Marnes-laCoquette, France) using the Mastermix PCR kit (Eurogentec) with 5 pmol of each primer and probe and 5 l of about 2 g of DNA into a 20-l final volume. M. oralis DSMZ ...

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Real-Time PCR Quantification of Methanobrevibacter oralis in Periodontitis

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“…periodontal disease. 20 Sixteen studies were selected [8][9][10][11][12][13][14][15]23,[26][27][28][29][30][31] and Methanobrevibacter, Methanosphaera, Methanosarcina, Thermoplasmata and Methanobacterium were considered the five most common genera of methanogenic Archaea present in the subgingival biofilm. In addition, the pooled prevalence of these studies showed that M. oralis was detected in 41% of the periodontitis subjects in comparison with only 6% of the periodontally healthy subjects (p < 0.05).…”

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confidence: 99%

Reduction in prevalence ofArchaeaafter periodontal therapy in subjects with generalized aggressive periodontitis

et al. 2013

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Background: There is evidence of a possible relationship between Archaea and periodontal disease; however, to date few studies have assessed the changes in prevalence of this domain after periodontal therapy. The aim of this randomized double-blind and placebo-controlled study was to assess if periodontal treatment with or without systemic antibiotic would change the prevalence of Archaea after periodontal therapy. Methods: Thirty subjects were randomly assigned to receive scaling and root planing (SRP) alone or combined with metronidazole (MTZ) + amoxicillin (AMX) for 14 days. Clinical and microbiological examinations were performed at baseline and at six months post-SRP. Nine subgingival plaque samples per subject were analysed for the presence of Archaea. Results: SRP alone or combined with MTZ + AMX significantly reduced the prevalence of subjects colonized by Archaea at six months post-therapy. However, no significant differences between treatment groups were observed (p > 0.05). Both therapies led to a statistically significant decrease in the mean percentage of sites colonized by Archaea (p < 0.05). A negative Spearman correlation was observed between the presence of Archaea and the mean clinical attachment gain at six months post-therapy (r 2 = -0.61; 95% CI -0.80--0.31; p = 0.003). Conclusions: SRP alone or combined with MTZ + AMX provides a similar reduction in the prevalence of Archaea in the subgingival biofilm of subjects with generalized aggressive periodontitis.

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“…Indeed, as a function of progressing disease, increasing numbers of Archaea (including Methanobrevibacter oralis) can be isolated from subgingival pockets. Despite the lack of any direct cause-effect in vivo studies assessing the role of Archaea in periodontitis, strikingly, these organisms have never been found in the subgingival microflora of healthy individuals or at healthy sites in patients with periodontitis (115,116). The difference between the onsets of chronic and localized aggressive forms of periodontitis is unclear at this time but is thought to result from a host genetic predisposition to colonization, a hyperaggressive immune response to oral bacteria involving increased levels of polymorphonuclear infiltrate, differences mediated by the microbial composition itself, or a combination of these effectors (44,58,84).…”

Section: Diseases Of the Oral Cavitymentioning

confidence: 99%

Polymicrobial Interactions: Impact on Pathogenesis and Human Disease

et al. 2012

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SUMMARY Microorganisms coexist in a complex milieu of bacteria, fungi, archaea, and viruses on or within the human body, often as multifaceted polymicrobial biofilm communities at mucosal sites and on abiotic surfaces. Only recently have we begun to appreciate the complicated biofilm phenotype during infection; moreover, even less is known about the interactions that occur between microorganisms during polymicrobial growth and their implications in human disease. Therefore, this review focuses on polymicrobial biofilm-mediated infections and examines the contribution of bacterial-bacterial, bacterial-fungal, and bacterial-viral interactions during human infection and potential strategies for protection against such diseases.

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Prevalence and molecular diversity of Archaea in subgingival pockets of periodontitis patients

Cited by 74 publications

References 24 publications

Real-Time PCR Quantification of Methanobrevibacter oralis in Periodontitis

Real-Time PCR Quantification of Methanobrevibacter oralis in Periodontitis

Reduction in prevalence ofArchaeaafter periodontal therapy in subjects with generalized aggressive periodontitis

Polymicrobial Interactions: Impact on Pathogenesis and Human Disease

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