1887

Abstract

The oral cavity has been implicated as a source of infection in childhood. Various PCR methods have been used to detect DNA in oral specimens with various detection rates reported. Such disparity in detection rates complicates the estimation of the true infection rate of in the oral cavity. In the present study, we constructed a novel PCR system for detection and used it to analyse oral specimens. Firstly, the nucleotide alignments of genes commonly used for detection were compared using the complete genome information for 48 strains registered in the GenBank database. Candidate primer sets with an estimated amplification size of approximately 300–400 bp were selected, and the specificity and sensitivity of the detection system using each primer set were evaluated. Five sets of primers targeting were considered appropriate, of which a single primer set was chosen for inclusion in the PCR system. The sensitivity of the system was considered appropriate and its detection limit established as one to ten cells per reaction. The novel PCR system was used to examine distribution in oral specimens (40 inflamed pulp tissues, 40 saliva samples) collected from Japanese children, adolescents and young adults. PCR analysis revealed that the detection rate of in inflamed pulp was 15 %, whereas no positive reaction was found in any of the saliva specimens. Taken together, our novel PCR system was found to be reliable for detecting . The results obtained showed that was detected in inflamed pulp but not saliva specimens, indicating that an infected root canal may be a reservoir for .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.079491-0
2015-01-01
2019-12-08
Loading full text...

Full text loading...

/deliver/fulltext/jmm/64/1/117.html?itemId=/content/journal/jmm/10.1099/jmm.0.079491-0&mimeType=html&fmt=ahah

References

  1. 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:, 1426–1430.[PubMed]
    [Google Scholar]
  2. Chung C., Olivares A., Torres E., Yilmaz O., Cohen H., Perez-Perez G.. ( 2010;). Diversity of VacA intermediate region among Helicobacter pylori strains from several regions of the world. . J Clin Microbiol 48:, 690–696. [CrossRef][PubMed]
    [Google Scholar]
  3. Córdova Espinoza M. G., González Vazquez R., Morales Mendez I., Ruelas Vargas C., Giono Cerezo S.. ( 2011;). Detection of the glmM gene in Helicobacter pylori isolates with a novel primer by PCR. . J Clin Microbiol 49:, 1650–1652. [CrossRef][PubMed]
    [Google Scholar]
  4. Dunn B. E., Cohen H., Blaser M. J.. ( 1997;). Helicobacter pylori. . Clin Microbiol Rev 10:, 720–741.[PubMed]
    [Google Scholar]
  5. Fennerty M. B.. ( 1994;). Helicobacter pylori. . Arch Intern Med 154:, 721–727. [CrossRef][PubMed]
    [Google Scholar]
  6. Furuta Y., Kawai M., Yahara K., Takahashi N., Handa N., Tsuru T., Oshima K., Yoshida M., Azuma T.. & other authors ( 2011;). Birth and death of genes linked to chromosomal inversion. . Proc Natl Acad Sci U S A 108:, 1501–1506. [CrossRef][PubMed]
    [Google Scholar]
  7. Goodwin C. S., Mendall M. M., Northfield T. C.. ( 1997;). Helicobacter pylori infection. . Lancet 349:, 265–269. [CrossRef][PubMed]
    [Google Scholar]
  8. Hirai Y., Hayashi S., Shimomura H., Oguma K., Yokota K.. ( 1999;). Association of Helicobacter pylori with gastroduodenal diseases. . Jpn J Infect Dis 52:, 183–197.[PubMed]
    [Google Scholar]
  9. Hirsch C., Tegtmeyer N., Rohde M., Rowland M., Oyarzabal O. A., Backert S.. ( 2012;). Live Helicobacter pylori in the root canal of endodontic-infected deciduous teeth. . J Gastroenterol 47:, 936–940. [CrossRef][PubMed]
    [Google Scholar]
  10. Jenkins W. M., Papapanou P. N.. ( 2001;). Epidemiology of periodontal disease in children and adolescents. . Periodontol 2000 26:, 16–32. [CrossRef][PubMed]
    [Google Scholar]
  11. Kawai M., Furuta Y., Yahara K., Tsuru T., Oshima K., Handa N., Takahashi N., Yoshida M., Azuma T.. & other authors ( 2011;). Evolution in an oncogenic bacterial species with extreme genome plasticity: Helicobacter pylori East Asian genomes. . BMC Microbiol 11:, 104. [CrossRef][PubMed]
    [Google Scholar]
  12. Kignel S., de Almeida Pina F., André E. A., Alves Mayer M. P., Birman E. G.. ( 2005;). Occurrence of Helicobacter pylori in dental plaque and saliva of dyspeptic patients. . Oral Dis 11:, 17–21. [CrossRef][PubMed]
    [Google Scholar]
  13. Lynch D. J., Michalek S. M., Zhu M., Drake D., Qian F., Banas J. A.. ( 2013;). Cariogenicity of Streptococcus mutans glucan-binding protein deletion mutants. . Oral Health Dent Manag 12:, 191–199.[PubMed]
    [Google Scholar]
  14. Mapstone N. P., Lynch D. A., Lewis F. A., Axon A. T., Tompkins D. S., Dixon M. F., Quirke P.. ( 1993;). Identification of Helicobacter pylori DNA in the mouths and stomachs of patients with gastritis using PCR. . J Clin Pathol 46:, 540–543. [CrossRef][PubMed]
    [Google Scholar]
  15. Matsuo T., Shirakami T., Ozaki K., Nakanishi T., Yumoto H., Ebisu S.. ( 2003;). An immunohistological study of the localization of bacteria invading root pulpal walls of teeth with periapical lesions. . J Endod 29:, 194–200. [CrossRef][PubMed]
    [Google Scholar]
  16. Miyabayashi H., Furihata K., Shimizu T., Ueno I., Akamatsu T.. ( 2000;). Influence of oral Helicobacter pylori on the success of eradication therapy against gastric Helicobacter pylori. . Helicobacter 5:, 30–37. [CrossRef][PubMed]
    [Google Scholar]
  17. Momtaz H., Souod N., Dabiri H., Sarshar M.. ( 2012;). Study of Helicobacter pylori genotype status in saliva, dental plaques, stool and gastric biopsy samples. . World J Gastroenterol 18:, 2105–2111. [CrossRef][PubMed]
    [Google Scholar]
  18. Morales-Espinosa R., Fernandez-Presas A., Gonzalez-Valencia G., Flores-Hernandez S., Delgado-Sapien G., Mendez-Sanchez J. L., Sanchez-Quezada E., Muñoz-Pérez L., Leon-Aguilar R.. & other authors ( 2009;). Helicobacter pylori in the oral cavity is associated with gastroesophageal disease. . Oral Microbiol Immunol 24:, 464–468. [CrossRef][PubMed]
    [Google Scholar]
  19. Nakano K., Inaba H., Nomura R., Nemoto H., Takeda M., Yoshioka H., Matsue H., Takahashi T., Taniguchi K.. & other authors ( 2006;). Detection of cariogenic Streptococcus mutans in extirpated heart valve and atheromatous plaque specimens. . J Clin Microbiol 44:, 3313–3317. [CrossRef][PubMed]
    [Google Scholar]
  20. Ogiwara H., Sugimoto M., Ohno T., Vilaichone R. K., Mahachai V., Graham D. Y., Yamaoka Y.. ( 2009;). Role of deletion located between the intermediate and middle regions of the Helicobacter pylori vacA gene in cases of gastroduodenal diseases. . J Clin Microbiol 47:, 3493–3500. [CrossRef][PubMed]
    [Google Scholar]
  21. Palmer R. J. Jr. ( 2014;). Composition and development of oral bacterial communities. . Periodontol 2000 64:, 20–39. [CrossRef][PubMed]
    [Google Scholar]
  22. Park C. Y., Kwak M., Gutierrez O., Graham D. Y., Yamaoka Y.. ( 2003;). Comparison of genotyping Helicobacter pylori directly from biopsy specimens and genotyping from bacterial cultures. . J Clin Microbiol 41:, 3336–3338. [CrossRef][PubMed]
    [Google Scholar]
  23. Patra R., Chattopadhyay S., De R., Datta S., Chowdhury A., Ramamurthy T., Nair G. B., Berg D. E., Mukhopadhyay A. K.. ( 2011;). Intact cag pathogenicity island of Helicobacter pylori without disease association in Kolkata, India. . Int J Med Microbiol 301:, 293–302. [CrossRef][PubMed]
    [Google Scholar]
  24. Peek R. M. Jr, Miller G. G., Tham K. T., Pérez-Pérez G. I., Cover T. L., Atherton J. C., Dunn G. D., Blaser M. J.. ( 1995;). Detection of Helicobacter pylori gene expression in human gastric mucosa. . J Clin Microbiol 33:, 28–32.[PubMed]
    [Google Scholar]
  25. Prasanthi C. H., Prasanthi N. L., Manikiran S. S., Rama Rao N.. ( 2011;). Focus on current trends in the treatment of Helicobacter pylori infection: An update. . Int J Pharm Sci Rev Res 9:, 42–51.
    [Google Scholar]
  26. Silva D. G., Stevens R. H., Macedo J. M., Albano R. M., Falabella M. E., Veerman E. C., Tinoco E. M.. ( 2009;). Detection of cytotoxin genotypes of Helicobacter pylori in stomach, saliva and dental plaque. . Arch Oral Biol 54:, 684–688. [CrossRef][PubMed]
    [Google Scholar]
  27. Silva D. G., Tinoco E. M., Rocha G. A., Rocha A. M., Guerra J. B., Saraiva I. E., Queiroz D. M.. ( 2010;). Helicobacter pylori transiently in the mouth may participate in the transmission of infection. . Mem Inst Oswaldo Cruz 105:, 657–660. [CrossRef][PubMed]
    [Google Scholar]
  28. Smith S. I., Oyedeji K. S., Arigbabu A. O., Cantet F., Megraud F., Ojo O. O., Uwaifo A. O., Otegbayo J. A., Ola S. O., Coker A. O.. ( 2004;). Comparison of three PCR methods for detection of Helicobacter pylori DNA and detection of cagA gene in gastric biopsy specimens. . World J Gastroenterol 10:, 1958–1960.[PubMed]
    [Google Scholar]
  29. Song Q., Haller B., Schmid R. M., Adler G., Bode G.. ( 1999;). Helicobacter pylori in dental plaque: a comparison of different PCR primer sets. . Dig Dis Sci 44:, 479–484. [CrossRef][PubMed]
    [Google Scholar]
  30. Tomb J. F., White O., Kerlavage A. R., Clayton R. A., Sutton G. G., Fleischmann R. D., Ketchum K. A., Klenk H. P., Gill S.. & other authors ( 1997;). The complete genome sequence of the gastric pathogen Helicobacter pylori. . Nature 388:, 539–547. [CrossRef][PubMed]
    [Google Scholar]
  31. Umeda M., Kobayashi H., Takeuchi Y., Hayashi J., Morotome-Hayashi Y., Yano K., Aoki A., Ohkusa T., Ishikawa I.. ( 2003;). High prevalence of Helicobacter pylori detected by PCR in the oral cavities of periodontitis patients. . J Periodontol 74:, 129–134. [CrossRef][PubMed]
    [Google Scholar]
  32. Wang J., Chi D. S., Laffan J. J., Li C., Ferguson D. A. Jr, Litchfield P., Thomas E.. ( 2002;). Comparison of cytotoxin genotypes of Helicobacter pylori in stomach and saliva. . Dig Dis Sci 47:, 1850–1856. [CrossRef][PubMed]
    [Google Scholar]
  33. Westblom T. U., Bhatt B. D.. ( 1999;). Diagnosis of Helicobacter pylori infection. . Curr Top Microbiol Immunol 241:, 215–235.[PubMed]
    [Google Scholar]
  34. Yip K., Smales R.. ( 2012;). Oral diagnosis and treatment planning: part 2. Dental caries and assessment of risk. . Br Dent J 213:, 59–66. [CrossRef][PubMed]
    [Google Scholar]
  35. Yu C., Abbott P. V.. ( 2007;). An overview of the dental pulp: its functions and responses to injury. . Aust Dent J 52: (Suppl S1), S4–S6. [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.079491-0
Loading
/content/journal/jmm/10.1099/jmm.0.079491-0
Loading

Data & Media loading...

Supplements

Supplementary Data



PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error