Current Microbiology

, Volume 62, Issue 1, pp 78–83 | Cite as

Simultaneous Amplification of Two Bacterial Genes: More Reliable Method of Helicobacter pylori Detection in Microbial Rich Dental Plaque Samples

  • Saima ChaudhryEmail author
  • Muhammad Idrees
  • Mateen Izhar
  • Arshad Kamal Butt
  • Ayyaz Ali Khan


Polymerase Chain reaction (PCR) assay is considered superior to other methods for detection of Helicobacter pylori (H. pylori) in oral cavity; however, it also has limitations when sample under study is microbial rich dental plaque. The type of gene targeted and number of primers used for bacterial detection in dental plaque samples can have a significant effect on the results obtained as there are a number of closely related bacterial species residing in plaque biofilm. Also due to high recombination rate of H. pylori some of the genes might be down regulated or absent. The present study was conducted to determine the frequency of H. pylori colonization of dental plaque by simultaneously amplifying two genes of the bacterium. One hundred dental plaque specimens were collected from dyspeptic patients before their upper gastrointestinal endoscopy and presence of H. pylori was determined through PCR assay using primers targeting two different genes of the bacterium. Eighty-nine of the 100 samples were included in final analysis. With simultaneous amplification of two bacterial genes 51.6% of the dental plaque samples were positive for H. pylori while this prevalence increased to 73% when only one gene amplification was used for bacterial identification. Detection of H. pylori in dental plaque samples is more reliable when two genes of the bacterium are simultaneously amplified as compared to one gene amplification only.


Polymerase Chain Reaction Polymerase Chain Reaction Assay Polymerase Chain Reaction Analysis Dental Plaque Gastric Biopsy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Lehours P, Yilmaz O (2007) Epidemiology of Helicobacter pylori infection. Helicobacter 12:1–3CrossRefPubMedGoogle Scholar
  2. 2.
    Azevedo NF, Huntington J, Goodman KJ (2009) The epidemiology of Helicobacter pylori and public health implications. Helicobacter 14:1–7CrossRefPubMedGoogle Scholar
  3. 3.
    Chaudhry S, Iqbal HA, Khan AA, Izhar M, Butt AK, Akhter MW, Izhar F, Mirza KM (2008) Helicobacter pylori in dental plaque and gastric mucosa: correlation revisited. J Pak Med Assoc 58:331–334PubMedGoogle Scholar
  4. 4.
    Butt AK, Khan AA, Khan AA, Izhar M, Alam A, Shah SW, Shafqat F (2002) Correlation of Helicobacter pylori in dental plaque and gastric mucosa of dyspeptic patients. J Pak Med Assoc 52:196–200PubMedGoogle Scholar
  5. 5.
    Jia CL, Jiang GS, Li CH, Li CR (2009) Effect of dental plaque control on infection of helicobacter pylori in gastric mucosa. J Periodontol 80:1606–1609CrossRefPubMedGoogle Scholar
  6. 6.
    Eskandari A, Mahmoodpoor A, Abolfazli N, Lafzi A (2009) Detection of Helicobacter pylori using PCR in dental plaque of patients with and without gastritis. Med Oral Patol Oral Cir Bucal Sept 21. (Epub ahead of print)Google Scholar
  7. 7.
    Medina ML, Medina MG, Martín GT, Picón SO, Bancalari A, Merino LA (2009) Molecular detection of Helicobacter pylori in oral samples from patients suffering digestive pathologies. Med Oral Patol Oral Cir Bucal Aug 13. (Epub ahead of print)Google Scholar
  8. 8.
    Liu Y, Yue H, Li A, Wang J, Jiang B, Zhang Y, Bai Y (2009) An epidemiologic study on the correlation between oral Helicobacter pylori and gastric H. pylori. Curr Microbiol 58:449–453CrossRefPubMedGoogle Scholar
  9. 9.
    Bürgers R, Schneider-Brachert W, Reischl U, Behr A, Hiller KA, Lehn N, Schmalz G, Ruhl S (2008) Helicobacter pylori in human oral cavity and stomach. Eur J Oral Sci 116:297–304CrossRefPubMedGoogle Scholar
  10. 10.
    Berroteran A, Perrone M, Correnti M, Cavazza ME, Tombazzi C, Goncalvez R, Lecuna V (2002) Detection of Helicobacter pylori DNA in the oral cavity and gastroduodenal system of a Venezuelan population. J Med Microbiol 51:764–770PubMedGoogle Scholar
  11. 11.
    Ogunbodede EO, Lawal OO, Lamikanra A, Okeke IN, Rotimi O, Rasheed AA (2002) Helicobacter pylori in the dental plaque and gastric mucosa of dyspeptic Nigerian patients. Trop Gastroenterol 23:127–133PubMedGoogle Scholar
  12. 12.
    Chitsazi MT, Fattahi E, Farahani RM, Fattahi S (2006) Helicobacter pylori in the dental plaque: is it of diagnostic value for gastric infection? Med Oral Patol Oral Cir Bucal 1:E325–E328Google Scholar
  13. 13.
    Kignel S, de Almeida Pina F, André EA, Alves Mayer MP, Birman EG (2005) Occurrence of Helicobacter pylori in dental plaque and saliva of dyspeptic patients. Oral Dis 11:17–21CrossRefPubMedGoogle Scholar
  14. 14.
    De Sousa L, Vásquez L, Velasco J, Parlapiano D (2006) Isolation of Helicobacter pylori in gastric mucosa, dental plaque and saliva in a population from the Venezuelan Andes. Invest Clin 47:109–116PubMedGoogle Scholar
  15. 15.
    Siddiq M, Rehman H, Mahmood A (2004) Evidence of Helicobacter pylori infection in dental plaque and gastric mucosa. J Coll Physicians Surg Pak 14:205–207PubMedGoogle Scholar
  16. 16.
    Song Q, Haller B, Schmid RM, Adler G, Bode G (1999) Helicobacter pylori in dental plaque: a comparison of different PCR primer sets. Dig Dis Sci 44:479–484CrossRefPubMedGoogle Scholar
  17. 17.
    Silva Rossi-Aguiar VP, Navarro-Rodriguez T, Mattar R, Siqueira de Melo Peres MP, Correa Barbuti R, Silva FM, Carrilho FJ, Eisig JN (2009) Oral cavity is not a reservoir for Helicobacter pylori in infected patients with functional dyspepsia. Oral Microbiol Immunol 24:255–259CrossRefPubMedGoogle Scholar
  18. 18.
    Olivier BJ, Bond RP, van Zyl WB, Delport M, Slavik T, Ziady C, Terhaar Sive Droste JS, Lastovica A, van der Merwe SW (2006) Absence of Helicobacter pylori within the oral cavities of members of a healthy South African community. J Clin Microbiol 44:635–636CrossRefPubMedGoogle Scholar
  19. 19.
    Bernander S, Dalén J, Gästrin B, Hedenborg L, Lamke LO, Ohrn R (1993) Absence of Helicobacter pylori in dental plaques in Helicobacter pylori positive dyspeptic patients. Eur J Clin Microbiol Infect Dis 12:282–285CrossRefPubMedGoogle Scholar
  20. 20.
    Sahin FI, Tinaz AC, Simşek IS, Menevşe S, Görgül A (2001) Detection of Helicobacter pylori in dental plaque and gastric biopsy samples of Turkish patients by PCR-RFLP. Acta Gastroenterol Belg 64:150–152PubMedGoogle Scholar
  21. 21.
    Dowsett SA, Kowolik MJ (2003) Oral Helicobacter pylori: can we stomach it? Crit Rev Oral Biol Med 14:226–233CrossRefPubMedGoogle Scholar
  22. 22.
    Andersen LP, Rasmussen L (2009) Helicobacter pylori coccoid forms and biofilm formation. FEMS Immunol Med Microbiol 56:112–115CrossRefPubMedGoogle Scholar
  23. 23.
    Marsh PD (2004) Dental plaque as a microbial biofilm. Caries Res 38:204–211CrossRefPubMedGoogle Scholar
  24. 24.
    Dong QJ, Wang Q, Xin YN, Li N, Xuan SY (2009) Comparative genomics of Helicobacter pylori. World J Gastroenterol 15:3984–3991CrossRefPubMedGoogle Scholar
  25. 25.
    Butt AK, Khan AA, Bedi R (1999) Helicobacter pylori in dental plaque of Pakistanis. J Int Acad Periodontol 1:78–82PubMedGoogle Scholar
  26. 26.
    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 74:129–134CrossRefPubMedGoogle Scholar
  27. 27.
    Song Q, Lange T, Spahr A, Adler G, Bode G (2000) Characteristic distribution pattern of Helicobacter pylori in dental plaque and saliva detected with nested PCR. J Med Microbiol 49:349–353PubMedGoogle Scholar
  28. 28.
    Nasrolahei M, Sharif M, Ahanjan M, Daryani A (2008) Prevalence of Helicobacter pylori in gastric mucosa and dental plaque in Sari, Northern Iran. J Chin Clin Med 7:383–388Google Scholar
  29. 29.
    Teoman I, Ozmeriç N, Ozcan G, Alaaddinoğlu E, Dumlu S, Akyön Y, Baloş K (2007) Comparison of different methods to detect Helicobacter pylori in the dental plaque of dyspeptic patients. Clin Oral Investig 11:201–205CrossRefPubMedGoogle Scholar
  30. 30.
    Sugimoto M, Wu JY, Abudayyeh S, Hoffman J, Brahem H, Al-Khatib K, Yamaoka Y, Graham DY (2009) Unreliability of results of PCR detection of Helicobacter pylori in clinical or environmental samples. J Clin Microbiol 47:738–742CrossRefPubMedGoogle Scholar
  31. 31.
    Goosen C, Theron J, Ntsala M, Maree FF, Olckers A, Botha SJ, Lastovica AJ, van der Merwe SW (2002) Evaluation of a novel heminested PCR assay based on the phosphoglucosamine mutase gene for detection of Helicobacter pylori gene in saliva and dental plaque. J Clin Microbiol 40:205–209CrossRefPubMedGoogle Scholar
  32. 32.
    Lundström AM, Bölin I (2000) A 26 kDa protein of Helicobacter pylori shows alkyl hydroperoxide reductase (AhpC) activity and the mono-cistronic transcription of the gene is affected by pH. Microb Pathog 29:257–266CrossRefPubMedGoogle Scholar
  33. 33.
    Liu H, Rehman A, Semino-Mora C, Doi SQ, Dubois A (2008) Specific and sensitive detection of H. pylori in biological specimens by real time RT-PCR and in situ hybridization. PLoS ONE 3:e2689. doi: 10.1371/journal/pone.0002689

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Saima Chaudhry
    • 1
    Email author
  • Muhammad Idrees
    • 2
  • Mateen Izhar
    • 3
  • Arshad Kamal Butt
    • 4
  • Ayyaz Ali Khan
    • 1
  1. 1.Department of Oral Health SciencesShaikh Zayed Medical ComplexLahorePakistan
  2. 2.Molecular Virology Lab, National Centre of Excellence in Molecular BiologyUniversity of the PunjabLahorePakistan
  3. 3.Department of Microbiology and VirologyShaikh Zayed Medical ComplexLahorePakistan
  4. 4.Department of Gastroenterology and HepatologyShaikh Zayed Medical ComplexLahorePakistan

Personalised recommendations