Detection of oral bacteria on the tongue dorsum using PCR amplification of 16S ribosomal RNA and its association with systemic disease in middle-aged and elderly patients

Su,Cheng-Yih; Shigeishi,Hideo; Nishimura,Rumi; Ohta,Kouji; Sugiyama,Masaru

doi:10.3892/br.2018.1175

Detection of oral bacteria on the tongue dorsum using PCR amplification of 16S ribosomal RNA and its association with systemic disease in middle-aged and elderly patients

Authors:
- Cheng-Yih Su
- Hideo Shigeishi
- Rumi Nishimura
- Kouji Ohta
- Masaru Sugiyama
View Affiliations

Affiliations: Department of Public Oral Health, Program of Oral Health Sciences, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734-8553, Japan, Department of Oral Epidemiology, Program of Oral Health Sciences, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734-8553, Japan, Department of Oral and Maxillofacial Surgery, Program of Dentistry, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734-8553, Japan
Published online on: November 26, 2018 https://doi.org/10.3892/br.2018.1175
Pages: 70-76

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The association between oral health and systemic disease is recognized in the literature. The present study aimed to clarify the association between oral bacteria on the tongue dorsum and factors associated with oral health and systemic disease in middle‑aged and elderly patients. The association between bacterial numbers, oral health status and systemic disease was preliminarily investigated in 70 patients (mean age, 69.5 years; range, 45‑92 years) who visited the Department of Oral Health, Hiroshima University Hospital (Hiroshima, Japan). The bacterial 16S ribosomal RNA gene was employed to quantitate bacterial numbers using real‑time polymerase chain reaction (PCR). PCR was also performed to detect the DNA of periodontal disease‑related bacteria. Oral bacterial numbers were marginally negatively correlated with moisture levels on the tongue surface [Spearman's rank correlation coefficient (R)=-0.131, P=0.28). Subjects with bleeding on probing (BOP) or a ≥4 mm probing depth (PD) exhibited higher Porphyromonas gingivalis (P. gingivalis)‑positive rates (50.0 and 51.1%, respectively) than those without BOP or a <4 mm PD (39.5 and 30.4%, respectively). Subjects with medical histories of hypertension, diabetes, stroke and heart disease exhibited a trend toward higher P. gingivalis‑positive rates than those without such disorders. These findings indicated that the tongue moisture level may be associated with bacterial numbers on the tongue surface, while P. gingivalis on the tongue surface may be associated with systemic and periodontal diseases. Further investigation in a larger number of participants is necessary to clarify the correlation between bacterial numbers and systemic disease.

View Figures

View References

1	Terpenning M: Geriatric oral health and pneumonia risk. Clin Infect Dis. 40:1807–1810. 2005. View Article : Google Scholar : PubMed/NCBI
2	Shigeishi H, Ohta K, Fujimoto S, Nakagawa T, Mizuta K, Ono S, Shimasue H, Ninomiya Y, Higashikawa K, Tada M, et al: Preoperative oral health care reduces postoperative inflammation and complications in oral cancer patients. Exp Ther Med. 12:1922–1928. 2016. View Article : Google Scholar : PubMed/NCBI
3	Turner MD and Ship JA: Dry mouth and its effects on the oral health of elderly people. J Am Dent Assoc. 138:15S–20S. 2007. View Article : Google Scholar : PubMed/NCBI
4	Kawano T, Shigeishi H, Fukada E, Yanagisawa T, Kuroda N, Takemoto T and Sugiyama M: Changes in bacterial number at different sites of oral cavity during perioperative oral care management in gastrointestinal cancer patients: Preliminary study. J Appl Oral Sci. 26:e201705162018. View Article : Google Scholar : PubMed/NCBI
5	Hayashida S, Funahara M, Sekino M, Yamaguchi N, Kosai K, Yanamoto S, Yanagihara K and Umeda M: The effect of tooth brushing, irrigation, and topical tetracycline administration on the reduction of oral bacteria in mechanically ventilated patients: A preliminary study. BMC Oral Health. 16:672016. View Article : Google Scholar : PubMed/NCBI
6	Kobayashi K, Ryu M, Izumi S, Ueda T and Sakurai K: Effect of oral cleaning using mouthwash and a mouth moisturizing gel on bacterial number and moisture level of the tongue surface of older adults requiring nursing care. Geriatr Gerontol Int. 17:116–121. 2017. View Article : Google Scholar : PubMed/NCBI
7	Danser MM, Gómez SM and Van der Weijden GA: Tongue coating and tongue brushing: A literature review. Int J Dent Hyg. 1:151–158. 2003. View Article : Google Scholar : PubMed/NCBI
8	Segata N, Haake SK, Mannon P, Lemon KP, Waldron L, Gevers D, Huttenhower C and Izard J: Composition of the adult digestive tract bacterial microbiome based on seven mouth surfaces, tonsils, throat and stool samples. Genome Biol. 13:R422012. View Article : Google Scholar : PubMed/NCBI
9	Horz HP, Vianna ME, Gomes BP and Conrads G: Evaluation of universal probes and primer sets for assessing total bacterial load in clinical samples: General implications and practical use in endodontic antimicrobial therapy. J Clin Microbiol. 43:5332–5337. 2005. View Article : Google Scholar : PubMed/NCBI
10	Suzuki N, Yoshida A and Nakano Y: Quantitative analysis of multi-species oral biofilms by TaqMan Real-Time PCR. Clin Med Res. 3:176–185. 2005. View Article : Google Scholar : PubMed/NCBI
11	Cutress TW, Ainamo J and Sardo-Infirri J: The community periodontal index of treatment needs (CPITN) procedure for population groups and individuals. Int Dent J. 37:222–233. 1987.PubMed/NCBI
12	Kembel SW, Wu M, Eisen JA and Green JL: Incorporating 16S gene copy number information improves estimates of microbial diversity and abundance. PLOS Comput Biol. 8:e10027432012. View Article : Google Scholar : PubMed/NCBI
13	Perisin M, Vetter M, Gilbert JA and Bergelson J: 16Stimator: Statistical estimation of ribosomal gene copy numbers from draft genome assemblies. ISME J. 10:1020–1024. 2016. View Article : Google Scholar : PubMed/NCBI
14	Yoshida A, Suzuki N, Nakano Y, Oho T, Kawada M and Koga T: Development of a 5 fluorogenic nuclease-based real-time PCR assay for quantitative detection of Actinobacillus actinomycetemcomitans Porphyromonas gingivalis. J Clin Microbiol. 41:863–866. 2003. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Yasukawa T, Ohmori M and Sato S: The relationship between physiologic halitosis and periodontopathic bacteria of the tongue and gingival sulcus. Odontology. 98:44–51. 2010. View Article : Google Scholar : PubMed/NCBI
17	Takahashi F, Koji T and Morita O: Oral dryness examinations: Use of an oral moisture checking device and the usefulness of an oral moisture checking device and a modified cotton method. Prosthodont Res Pract. 5:26–30. 2006. View Article : Google Scholar
18	Winkel EG, Roldán S, Van Winkelhoff AJ, Herrera D and Sanz M: Clinical effects of a new mouthrinse containing chlorhexidine, cetylpyridinium chloride and zinc-lactate on oral halitosis. A dual-center, double-blind placebo-controlled study. J Clin Periodontol. 30:300–306. 2003. View Article : Google Scholar : PubMed/NCBI
19	Socransky SS, Haffajee AD, Cugini MA, Smith C and Kent RL Jr: Microbial complexes in subgingival plaque. J Clin Periodontol. 25:134–144. 1998. View Article : Google Scholar : PubMed/NCBI
20	Veal DA, Deere D, Ferrari B, Piper J and Attfield PV: Fluorescence staining and flow cytometry for monitoring microbial cells. J Immunol Methods. 243:191–210. 2000. View Article : Google Scholar : PubMed/NCBI
21	Socransky SS, Smith C, Martin L, Paster BJ, Dewhirst FE and Levin AE: ‘Checkerboard’ DNA-DNA hybridization. Biotechniques. 17:788–792. 1994.PubMed/NCBI
22	Hamada R, Suehiro J, Nakano M, Kikutani T and Konishi K: Development of rapid oral bacteria detection apparatus based on dielectrophoretic impedance measurement method. IET Nanobiotechnol. 5:25–31. 2011. View Article : Google Scholar : PubMed/NCBI
23	Genco RJ and Borgnakke WS: Risk factors for periodontal disease. Periodontol 2000. 62:59–94. 2013. View Article : Google Scholar : PubMed/NCBI
24	Nishimura F, Iwamoto Y, Mineshiba J, Shimizu A, Soga Y and Murayama Y: Periodontal disease and diabetes mellitus: The role of tumor necrosis factor-alpha in a 2-way relationship. J Periodontol. 74:97–102. 2003. View Article : Google Scholar : PubMed/NCBI
25	Chigasaki O, Takeuchi Y, Aoki A, Sasaki Y, Mizutani K, Aoyama N, Ikeda Y, Gokyu M, Umeda M, Ishikawa I, et al: A cross-sectional study on the periodontal status and prevalence of red complex periodontal pathogens in a Japanese population. J Oral Sci. 60:293–303. 2018. View Article : Google Scholar : PubMed/NCBI
26	Faveri M, Feres M, Shibli JA, Hayacibara RF, Hayacibara MM and de Figueiredo LC: Microbiota of the dorsum of the tongue after plaque accumulation: An experimental study in humans. J Periodontol. 77:1539–1546. 2006. View Article : Google Scholar : PubMed/NCBI
27	Ford PJ, Gemmell E, Hamlet SM, Hasan A, Walker PJ, West MJ, Cullinan MP and Seymour GJ: Cross-reactivity of GroEL antibodies with human heat shock protein 60 and quantification of pathogens in atherosclerosis. Oral Microbiol Immunol. 20:296–302. 2005. View Article : Google Scholar : PubMed/NCBI
28	Seymour GJ, Ford PJ, Cullinan MP, Leishman S and Yamazaki K: Relationship between periodontal infections and systemic disease. Clin Microbiol Infect. 13 Suppl 4:3–10. 2007. View Article : Google Scholar : PubMed/NCBI
29	Li H, Yang H, Ding Y, Aprecio R, Zhang W, Wang Q and Li Y: Experimental periodontitis induced by Porphyromonas gingivalis does not alter the onset or severity of diabetes in mice. J Periodontal Res. 48:582–590. 2013. View Article : Google Scholar : PubMed/NCBI
30	Meyer-Bäumer A, Eick S, Mertens C, Uhlmann L, Hagenfeld D, Eickholz P, Kim TS and Cosgarea R: Periodontal pathogens and associated factors in aggressive periodontitis: Results 5-17 years after active periodontal therapy. J Clin Periodontol. 41:662–672. 2014. View Article : Google Scholar : PubMed/NCBI