Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JCM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Journal of Clinical Microbiology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JCM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Bacteriology

Detection of Multiple Pathogenic Species in Saliva Is Associated with Periodontal Infection in Adults

Susanna Paju, Pirkko J. Pussinen, Liisa Suominen-Taipale, Mari Hyvönen, Matti Knuuttila, Eija Könönen
Susanna Paju
1Institute of Dentistry, University of Helsinki and Department of Oral and Maxillofacial Diseases, Helsinki University Central Hospital, Helsinki, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: Susanna.Paju@helsinki.fi
Pirkko J. Pussinen
1Institute of Dentistry, University of Helsinki and Department of Oral and Maxillofacial Diseases, Helsinki University Central Hospital, Helsinki, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Liisa Suominen-Taipale
2Department of Health and Functional Capacity
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mari Hyvönen
3Department of Bacterial and Inflammatory Diseases, National Public Health Institute (KTL), Helsinki, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Matti Knuuttila
4Institute of Dentistry, University of Oulu, Oulu, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Eija Könönen
3Department of Bacterial and Inflammatory Diseases, National Public Health Institute (KTL), Helsinki, Finland
5Institute of Dentistry, University of Turku, Turku, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/JCM.01824-08
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

ABSTRACT

We investigated whether certain bacterial species and their combinations in saliva can be used as markers for periodontitis. In 1,198 subjects, the detection of multiple species, rather than the presence of a certain pathogen, in saliva was associated with periodontitis as determined by the number of teeth with deepened periodontal pockets.

Periodontitis, infection of the tooth-supporting tissues, results from the accumulation of pathogenic bacterial plaque at and below the gingival margin (12). The composition of the dental plaque community plays a central role in the etiology of periodontitis (7, 11, 15). The major periodontal pathogens are Aggregatibacter (formerly Actinobacillus) actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia (previously forsythensis), Campylobacter rectus, and Treponema denticola (2, 7, 22). In subgingival plaque, P. gingivalis, T. forsythia, and T. denticola have the strongest relation to periodontal tissue destruction (16).

The presence of pathogens in subgingival sites of early and advanced periodontitis and in the healthy periodontium has been studied previously (1, 6, 16, 17, 22), while the natural carriage rates of periodontal pathogens in saliva are hardly known. Recently, we showed in a population-based study of Finnish adults that distinct periodontal bacteria have different carriage profiles depending on the age, educational level, and periodontal status of the subjects (9). The salivary carriage of periodontal pathogens proved to be common: out of the six examined periodontal pathogens, at least one was found in 88% of the subjects (9). Since major periodontal bacteria are commonly found in adults, a combination of pathogenic bacteria in saliva may represent a marker for disease. The objective of the present study was to investigate whether saliva, an easily and noninvasively collectable specimen material, can be used for diagnostic purposes of periodontitis.

The study subjects belong to a national population-based “Health 2000 Health Examination Survey,” coordinated by the National Public Health Institute (KTL), Finland (http://www.ktl.fi/health2000/index.uk.html/ ). All protocols were approved by the institutional ethic committees. Methods and patient recruitment have been previously published (9). The present study includes data for 1,198 dentate subjects, belonging to the southern Finland sample (n = 2,616), from whom both clinical data on oral health examination and microbiological data on salivary bacteria were available (9).

The number of teeth (all teeth and tooth remnants) and the number of periodontally diseased teeth (excluding third molars) determined by having probing pocket depths (PPDs) of ≥4 mm and ≥6 mm were recorded by a specially trained dentist. Smoking history and level of education were gathered by interviews (9). Bacterial DNA from saliva samples was extracted (9), and PCR detection for six periodontal pathogens, A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythia, C. rectus, and T. denticola, was performed using species-specific primers (3, 13, 20) as previously described.

Due to the skewed distribution of the outcome variables, a nonparametric test (Kruskal-Wallis analysis of variance) was used to analyze differences of the means in different numbers of pathogens. Relative risk (RR) and 95% confidence interval (95% CI) were estimated using Poisson regression models. The outcome variables included the number of teeth with PPDs of ≥4 mm and ≥6 mm. The number of species, presence of pathogens, and their various combinations were used as independent variables in addition to the age, sex, smoking history, and level of education of the subjects. The SUDAAN statistical package was used in the analyses to take into account two-stage cluster sampling.

Table 1 shows the characteristics of 1,198 study subjects. The number of the six studied periodontal species in saliva was associated with the number of teeth with PPDs of both ≥4 mm and ≥6 mm (P < 0.001) (Table 2). Among women who had never smoked, the higher number of pathogens was associated with the higher number of teeth with PPDs of ≥4 mm (data not shown). No such clear observations were made with men and women who smoked daily. Figure 1 shows percentages of subjects with certain bacterial species or different bacterial combinations. The associations between the presence of certain pathogens, alone or in any combination, and the number of teeth with deepened pockets are shown in Table 3. After adjustment, the carriage of P. gingivalis, despite the presence of other species, was significantly associated with the presence of PPDs of ≥6 mm (Table 3). The association of specific bacterial combinations (of two or three pathogens) with PPDs of ≥4 mm (Fig. 2A) was similar to the results with PPDs of ≥6 mm (Fig. 2B). Several combinations of four, five, and six pathogens were significantly associated with the occurrence of deepened pockets but were left out because the number of bacterial species, rather than the presence of certain species, proved to be important.

For the first time, in the present sample of 1,198 dentate Finnish adults, we report that the salivary carriage of multiple periodontal bacterial species is associated with periodontitis at the population level. Saliva is a representative diagnostic specimen for an overall view of the oral microbiota, since bacteria from various sites and surfaces of the oral cavity are found in saliva and mouth rinses (4, 8, 10, 19, 21). For example, A. actinomycetemcomitans has been detected in unstimulated saliva with no statistical difference to pooled subgingival samples (5). Subgingival curette sampling is a reproducible and reliable method for studying proportions of bacteria in periodontal biofilms (18). However, this technique requires a person with periodontal education/experience for selecting subgingival sites representative of periodontal status, whereas saliva can easily and less time-consumingly be collected at a dental hygienist's or nurse's appointment. As an easy and noninvasive specimen, saliva offers an excellent sample material for large population-based studies of periodontal health or carriage of periodontal pathogens.

The detection of multiple pathogenic species in saliva, rather than the presence of any single pathogen in saliva, was associated with periodontitis in our study. Although no specific combination was significantly more disease linked than others, A. actinomycetemcomitans, P. gingivalis, T. forsythia, and T. denticola, species that have been previously shown to have the strongest relationship to periodontal breakdown (2, 16), were also major players in the present bacterial combinations associated with the disease. Certain combinations in saliva were associated with the number of teeth with deepened pockets but not as strongly as has been reported for subgingival samples (3, 6, 16, 19, 22). This may be partially explained by the geographic location; subgingival microbial profiles have been found to differ in subjects from Europe and North and South America (6). In a recent study using a multiplex PCR method for detecting the subgingival presence of A. actinomycetemcomitans, P. gingivalis, and T. forsythia in periodontitis patients, subjects with a single pathogen had more severe disease than subjects with two or three pathogens, which suggests that both positive and negative bacterial interactions are important in periodontal biofilms (14).

We aimed to find out if saliva can be used for diagnostic purposes of periodontitis. Salivary sampling and PCR technique allow rapid identification of periodontal bacteria. In our study population, however, there were many distinctions between subjects of different ages, sexes, and behavioral habits such as smoking, and no specific disease marker could be established. The associations found in our cross-sectional study between the number of species and periodontal pockets were strong, however, suggesting possible predictive markers for periodontitis and encouraging further longitudinal studies. The present study in an adult population indicated that, rather than the presence of certain periodontal pathogens or specific combinations, the number of pathogenic species in saliva associates with clinical signs of periodontitis.

FIG. 1.
  • Open in new tab
  • Download powerpoint
FIG. 1.

Percentage of subjects with periodontal pathogens or different bacterial combinations in saliva in the study population (n = 1,198). Studied bacterial species are Aggregatibacter actinomycetemcomitans (Aa), Campylobacter rectus (Cr), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Treponema denticola (Td), and Tannerella forsythia (Tf).

FIG. 2.
  • Open in new tab
  • Download powerpoint
FIG. 2.

RR with 95% CI for the presence of teeth with PPDs of ≥4 mm (A) and ≥6 mm (B) in combinations of two or three pathogens in saliva (n = 1,198). Adjustments were made for the age, sex, education, number of teeth, and smoking habits of subjects. Studied bacterial species are as described in the legend to Fig. 1.

View this table:
  • View inline
  • View popup
TABLE 1.

Basic characteristics of the study subjects (n = 1,198)

View this table:
  • View inline
  • View popup
TABLE 2.

Distribution of subjects (n = 1,198) and the mean number of teeth with deepened pockets by the number of periodontal speciesc

View this table:
  • View inline
  • View popup
TABLE 3.

The relation of occurrence of pathogenic species with teeth with periodontal pockets explained by means of Poisson regression models in study subjects (n = 1,198)

ACKNOWLEDGMENTS

We thank the Health 2000 organization. Tiina Karvonen and Pirjo Nurmi are acknowledged for technical assistance.

Bacterial work has received financial support from the Academy of Finland (grant 78443 to E.K., grant 209152 to S.P., and grants 211129 and 118391 to P.J.P.). The oral health examination was partly supported by the Finnish Dental Society Apollonia and the Finnish Dental Association.

FOOTNOTES

    • Received 20 September 2008.
    • Returned for modification 1 November 2008.
    • Accepted 7 November 2008.
  • Copyright © 2009 American Society for Microbiology

REFERENCES

  1. 1.↵
    Aas, J. A., B. J. Paster, L. N. Stokes, I. Olsen, and F. E. Dewhirst. 2005. Defining the normal bacterial flora of the oral cavity. J. Clin. Microbiol.43:5721-5732.
    OpenUrlAbstract/FREE Full Text
  2. 2.↵
    American Academy of Periodontology. 1996. Consensus report. Periodontal diseases: pathogenesis and microbial factors. Ann. Periodontol.1:926-932.
    OpenUrlCrossRefPubMed
  3. 3.↵
    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.
    OpenUrlCrossRefPubMedWeb of Science
  4. 4.↵
    Boutaga, K., P. H. Savelkoul, E. G. Winkel, and A. J. van Winkelhoff. 2007. Comparison of subgingival bacterial sampling with oral lavage for detection and quantification of periodontal pathogens by real-time polymerase chain reaction. J. Periodontol.78:79-86.
    OpenUrlCrossRefPubMedWeb of Science
  5. 5.↵
    Cortelli, S. C., M. Feres, A. A. Rodrigues, D. R. Aquino, J. A. Shibli, and J. R. Cortelli. 2005. Detection of Actinobacillus actinomycetemcomitans in unstimulated saliva of patients with chronic periodontitis. J. Periodontol.76:204-209.
    OpenUrlCrossRefPubMed
  6. 6.↵
    Haffajee, A. D., A. Bogren, H. Hasturk, M. Feres, N. J. Lopez, and S. S. Socransky. 2004. Subgingival microbiota of chronic periodontitis subjects from different geographic locations. J. Clin. Periodontol.31:996-1002.
    OpenUrlCrossRefPubMedWeb of Science
  7. 7.↵
    Haffajee, A. D., and S. S. Socransky. 1994. Microbial etiological agents of destructive periodontal diseases. Periodontol. 20005:78-111.
    OpenUrlCrossRef
  8. 8.↵
    Könönen, E., H. Jousimies-Somer, and S. Asikainen. 1994. The most frequently isolated gram-negative anaerobes in saliva and subgingival samples taken from young women. Oral Microbiol. Immunol.9:126-128.
    OpenUrlPubMed
  9. 9.↵
    Könönen, E., S. Paju, P. J. Pussinen, M. Hyvönen, P. Di Tella, L. Suominen-Taipale, and M. Knuuttila. 2007. Population-based study of salivary carriage of periodontal pathogens in adults. J. Clin. Microbiol.45:2446-2451.
    OpenUrlAbstract/FREE Full Text
  10. 10.↵
    Mager, D. L., A. D. Haffajee, and S. S. Socransky. 2003. Effects of periodontitis and smoking on the microbiota of oral mucous membranes and saliva in systemically healthy subjects. J. Clin. Periodontol.30:1031-1037.
    OpenUrlCrossRefPubMed
  11. 11.↵
    Marsh, P. D. 2003. Are dental diseases examples of ecological catastrophes? Microbiology149:279-294.
    OpenUrlCrossRefPubMedWeb of Science
  12. 12.↵
    Pihlstrom, B. L., B. S. Michalowicz, and N. W. Johnson. 2005. Periodontal diseases. Lancet366:1809-1820.
    OpenUrlCrossRefPubMedWeb of Science
  13. 13.↵
    Premaraj, T., N. Kato, K. Fukui, H. Kato, and K. Watanabe. 1999. Use of PCR and sodium dodecyl sulfate-polyacrylamide gel electrophoresis techniques for differentiation of Prevotella intermedia sensu stricto and Prevotella nigrescens. J. Clin. Microbiol.37:1057-1061.
    OpenUrlAbstract/FREE Full Text
  14. 14.↵
    Ready, D., F. D'Aiuto, D. A. Spratt, J. Suvan, M. S. Tonetti, and M. Wilson. 2008. Disease severity associated with the presence in subgingival plaque of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Tannerella forsythia, singularly or in combination, as detected by nested multiplex PCR. J. Clin. Microbiol.46:3380-3383.
    OpenUrlAbstract/FREE Full Text
  15. 15.↵
    Socransky, S. S., and A. D. Haffajee. 2005. Periodontal microbial ecology. Periodontol. 200038:135-187.
    OpenUrlCrossRef
  16. 16.↵
    Socransky, S. S., A. D. Haffajee, M. A. Cugini, C. Smith, and R. L. Kent, Jr. 1998. Microbial complexes in subgingival plaque. J. Clin. Periodontol.25:134-144.
    OpenUrlCrossRefPubMedWeb of Science
  17. 17.↵
    Tanner, A. C., R. Kent, Jr., E. Kanasi, S. C. Lu, B. J. Paster, S. T. Sonis, L. A. Murray, and T. E. Van Dyke. 2007. Clinical characteristics and microbiota of progressing slight chronic periodontitis in adults. J. Clin. Periodontol.34:917-930.
    OpenUrlCrossRefPubMed
  18. 18.↵
    Teles, F. R., A. D. Haffajee, and S. S. Socransky. 2008. The reproducibility of curet sampling of subgingival biofilms. J. Periodontol.79:705-713.
    OpenUrlCrossRefPubMed
  19. 19.↵
    Timmerman, M. F., G. A. Van der Weijden, S. Armand, F. Abbas, E. G. Winkel, A. J. van Winkelhoff, and U. Van der Velden. 1998. Untreated periodontal disease in Indonesian adolescents. Clinical and microbiological baseline data. J. Clin. Periodontol.25:215-224.
    OpenUrlCrossRefPubMedWeb of Science
  20. 20.↵
    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.
    OpenUrlAbstract/FREE Full Text
  21. 21.↵
    Umeda, M., A. Contreras, C. Chen, I. Bakker, and J. Slots. 1998. The utility of whole saliva to detect the oral presence of periodontopathic bacteria. J. Periodontol.69:828-833.
    OpenUrlCrossRefPubMedWeb of Science
  22. 22.↵
    van Winkelhoff, A. J., B. G. Loos, W. A. van der Reijden, and U. van der Velden. 2002. Porphyromonas gingivalis, Bacteroides forsythus and other putative periodontal pathogens in subjects with and without periodontal destruction. J. Clin. Periodontol.29:1023-1028.
    OpenUrlCrossRefPubMedWeb of Science
PreviousNext
Back to top
Download PDF
Citation Tools
Detection of Multiple Pathogenic Species in Saliva Is Associated with Periodontal Infection in Adults
Susanna Paju, Pirkko J. Pussinen, Liisa Suominen-Taipale, Mari Hyvönen, Matti Knuuttila, Eija Könönen
Journal of Clinical Microbiology Jan 2009, 47 (1) 235-238; DOI: 10.1128/JCM.01824-08

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Journal of Clinical Microbiology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Detection of Multiple Pathogenic Species in Saliva Is Associated with Periodontal Infection in Adults
(Your Name) has forwarded a page to you from Journal of Clinical Microbiology
(Your Name) thought you would be interested in this article in Journal of Clinical Microbiology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Detection of Multiple Pathogenic Species in Saliva Is Associated with Periodontal Infection in Adults
Susanna Paju, Pirkko J. Pussinen, Liisa Suominen-Taipale, Mari Hyvönen, Matti Knuuttila, Eija Könönen
Journal of Clinical Microbiology Jan 2009, 47 (1) 235-238; DOI: 10.1128/JCM.01824-08
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

bacteria
biodiversity
Chronic Periodontitis
saliva

Related Articles

Cited By...

About

  • About JCM
  • Editor in Chief
  • Board of Editors
  • Editor Conflicts of Interest
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Resources for Clinical Microbiologists
  • Ethics
  • Contact Us

Follow #JClinMicro

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

 

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0095-1137; Online ISSN: 1098-660X