Detection of Actinobacillus actinomycetemcomitans DNA in Patients with Partial and Complete Dentures by Real-Time PCR

Enis Veseli, Gloria Staka

International Journal of Biomedicine. 2023;13(1):141-145.
DOI: 10.21103/Article13(1)_OA20
Originally published March 3, 2023


Background: The purpose of the present study was to detect Actinobacillus actinomycetemcomitans (Aa) using RT-PCR in patients with complete and partial edentulism before (T0) and three months after (T3) treatment with removable partial dentures (RPD) and complete dentures (CD), respectively, to compare the data between these two research groups.
Methods and Results: The sample comprised 60 patients: 33 men and 27 women, aged 48 to 76 years. The patients were divided into two groups. Group 1 included 30 patients with partial edentulism who were treated with RPD. Group 2 included 30 patients with complete edentulism who were treated with CD. The samples from Group 1 were taken from the gingival sulcus of the abutment teeth by means of sterile paper points. For Group 2, the samples were taken with a sterile swab from the dorsum of the tongue. The samples were taken in T0 and T3 intervals.  To detect Aa DNA, we used RT-PCR and ParodontoScreen REAL-TIME PCR Detection Kit (DNA-TECHNOLOGY). Bacterial load levels of species were conventionally represented in logarithm (Lg) of genome equivalents per sample. The results were also presented in three ranges depending on the level of bacterial load: normal (< 4.0Lg), light/moderate (≥4.0 Lg), and severe (>5.0 Lg).
The study found a significant difference in the amount of Aa between the T0 and T3 intervals only in patients treated with RPD (0.87±1.58 Lg vs. 1.28 ±1.96 Lg, P=0.004). Patients treated with CD, however, did not differ significantly in the amount of Aa between the T0 and T3 intervals (0.03±0.16 Lg vs. 0). The average bacterial load in patients with RPD was significantly higher than in those with CD three months after treatment (P=0.02) (Table 2). The range of bacterial load levels in the groups is presented in Table 3. Of the 30 patients with RPD, 2(6.7%) had a severe range, 2(6.7%) had a mild/moderate range, and 26(86.7%) had a normal range. The 30 CD patients all had a normal range. There was no significant difference in the prevalence range of bacterial load level with Aa between groups (Fisher's Exact Test = 3.537, P=0.113) / Monte Carlo Sig. (2-sided) / 0.105–0.121). However, in general, RPD causes a significant increase in Aa, so the level of periodontal pathogens may be higher in RPD patients than in CD patients.

Actinobacillus actinomycetemcomitans • real-time PCR • dentures
  1. Zhang JS, Chu CH, Yu OY. Oral Microbiome and Dental Caries Development. Dent J (Basel). 2022 Sep 30;10(10):184. doi: 10.3390/dj10100184. PMID: 36285994; PMCID: PMC9601200.
  2. Di Stefano M, Polizzi A, Santonocito S, Romano A, Lombardi T, Isola G. Impact of Oral Microbiome in Periodontal Health and Periodontitis: A Critical Review on Prevention and Treatment. Int J Mol Sci. 2022 May 5;23(9):5142. doi: 10.3390/ijms23095142. PMID: 35563531; PMCID: PMC9103139.
  3. Peng X, Cheng L, You Y, Tang C, Ren B, Li Y, Xu X, Zhou X. Oral microbiota in human systematic diseases. Int J Oral Sci. 2022 Mar 2;14(1):14. doi: 10.1038/s41368-022-00163-7. PMID: 35236828; PMCID: PMC8891310.
  4. Monteiro DR, de Souza Batista VE, Caldeirão ACM, Jacinto RC, Pessan JP. Oral prosthetic microbiology: aspects related to the oral microbiome, surface properties, and strategies for controlling biofilms. Biofouling. 2021 Apr;37(4):353-371. doi: 10.1080/08927014.2021.1912741. Epub 2021 Jun 17. PMID: 34139899.
  5. Sartawi SY, Abu-Hammad S, A Salim N, Al-Omoush S. Denture Stomatitis Revisited: A Summary of Systematic Reviews in the Past Decade and Two Case Reports of Papillary Hyperplasia of Unusual Locations. Int J Dent. 2021 Oct 13;2021:7338143. doi: 10.1155/2021/7338143. PMID: 34691183; PMCID: PMC8528609.
  6. Yasui M, Ryu M, Sakurai K, Ishihara K. Colonisation of the oral cavity by periodontopathic bacteria in complete denture wearers. Gerodontology. 2012 Jun;29(2):e494-502. doi: 10.1111/j.1741-2358.2011.00506.x. Epub 2011 Sep 19. PMID: 21929616.
  7. Cortelli JR, Aquino DR, Cortelli SC, Nobre Franco GC, Fernandes CB, Roman-Torres CV, Costa FO. Detection of periodontal pathogens in oral mucous membranes of edentulous individuals. J Periodontol. 2008 Oct;79(10):1962-5. doi: 10.1902/jop.2008.080092. PMID: 18834252.
  8. Fernandes CB, Aquino DR, Franco GC, Cortelli SC, Costa FO, Cortelli JR. Do elderly edentulous patients with a history of periodontitis harbor periodontal pathogens? Clin Oral Implants Res. 2010 Jun;21(6):618-23. doi: 10.1111/j.1600-0501.2009.01892.x. Epub 2010 Mar 11. PMID: 20337663.
  9. Mine K, Fueki K, Igarashi Y. Microbiological risk for periodontitis of abutment teeth in patients with removable partial dentures. J Oral Rehabil. 2009 Sep;36(9):696-702. doi: 10.1111/j.1365-2842.2009.01982.x. Epub 2009 Jul 23. PMID: 19650858.
  10. Ozuna H, Snider I, Belibasakis GN, Oscarsson J, Johansson A, Uriarte SM. Aggregatibacter actinomycetemcomitans and Filifactor alocis: Two exotoxin-producing oral pathogens. Front Oral Health. 2022 Aug 15;3:981343. doi: 10.3389/froh.2022.981343. PMID: 36046121; PMCID: PMC9420871.
  11. Oscarsson J, Claesson R, Lindholm M, Höglund Åberg C, Johansson A. Tools of Aggregatibacter actinomycetemcomitans to Evade the Host Response. J Clin Med. 2019 Jul 22;8(7):1079. doi: 10.3390/jcm8071079. PMID: 31336649; PMCID: PMC6678183.
  12. DNA-Technology. ParodontoScreen [Internet]. Online information available at: (Accessed 21 Dec 2022).
  13. Van der Weijden F, Rijnen M, Valkenburg C. Comparison of three qPCR-based commercial tests for detection of periodontal pathogens. Sci Rep. 2021 Mar 17;11(1):6141. doi: 10.1038/s41598-021-85305-3. PMID: 33731742; PMCID: PMC7969924.
  14. O'Donnell LE, Robertson D, Nile CJ, Cross LJ, Riggio M, Sherriff A, Bradshaw D, Lambert M, Malcolm J, Buijs MJ, Zaura E, Crielaard W, Brandt BW, Ramage G. The Oral Microbiome of Denture Wearers Is Influenced by Levels of Natural Dentition. PLoS One. 2015 Sep 14;10(9):e0137717. doi: 10.1371/journal.pone.0137717. PMID: 26368937; PMCID: PMC4569385.
  15. Gazdeck RK, Fruscione SR, Adami GR, Zhou Y, Cooper LF, Schwartz JL. Diversity of the oral microbiome between dentate and edentulous individuals. Oral Dis. 2019 Apr;25(3):911-918. doi: 10.1111/odi.13039. Epub 2019 Feb 10. PMID: 30657624.
  16. Sachdeo A, Haffajee AD, Socransky SS. Biofilms in the edentulous oral cavity. J Prosthodont. 2008 Jul;17(5):348-56. doi: 10.1111/j.1532-849X.2008.00301.x. PMID: 18355168.
  17. de Waal YC, Winkel EG, Raangs GC, van der Vusse ML, Rossen JW, van Winkelhoff AJ. Changes in oral microflora after full-mouth tooth extraction: a prospective cohort study. J Clin Periodontol. 2014 Oct;41(10):981-9. doi: 10.1111/jcpe.12297. Epub 2014 Sep 4. PMID: 25065274.
  18. Danser MM, van Winkelhoff AJ, de Graaff J, Loos BG, van der Velden U. Short-term effect of full-mouth extraction on periodontal pathogens colonizing the oral mucous membranes. J Clin Periodontol. 1994 Aug;21(7):484-9. doi: 10.1111/j.1600-051x.1994.tb00412.x. PMID: 7929861.
  19. Srinivas S, Ravi MB, Ram K, Jaishankar Homberhalli HP, Nagaraja MS, Gowrav MP, Ramesh K. Antibacterial Efficacy of Hubballi Propolis against Aggregatibacter Actinomycetemcomitans One of the Major Causative Organisms of Perimplantitis: An In vitro Study. J Pharm Bioallied Sci. 2022 Jul;14(Suppl 1):S595-S599. doi: 10.4103/jpbs.jpbs_653_21. Epub 2022 Jul 13. PMID: 36110765; PMCID: PMC9469338.
  20. Andjelkovic M, Sojic LT, Lemic AM, Nikolic N, Kannosh IY, Milasin J. Does the Prevalence of Periodontal Pathogens Change in Elderly Edentulous Patients after Complete Denture Treatment? J Prosthodont. 2017 Jul;26(5):364-369. doi: 10.1111/jopr.12402. Epub 2015 Nov 30. PMID: 26619204.
  21. Costa L, do Nascimento C, de Souza VO, Pedrazzi V. Microbiological and clinical assessment of the abutment and non-abutment teeth of partial removable denture wearers. Arch Oral Biol. 2017 Mar; 75:74-80. doi: 10.1016/j.archoralbio.2016.11.002. Epub 2016 Nov 3. PMID: 27825678.
  22. Zaura E, Keijser BJ, Huse SM, Crielaard W. Defining the healthy "core microbiome" of oral microbial communities. BMC Microbiol. 2009 Dec 15;9:259. doi: 10.1186/1471-2180-9-259. PMID: 20003481; PMCID: PMC2805672.
  23. Al-Ahmad A, Roth D, Wolkewitz M, Wiedmann-Al-Ahmad M, Follo M, Ratka-Krüger P, Deimling D, Hellwig E, Hannig C. Change in diet and oral hygiene over an 8-week period: effects on oral health and oral biofilm. Clin Oral Investig. 2010 Aug;14(4):391-6. doi: 10.1007/s00784-009-0318-9. Epub 2009 Jul 22. PMID: 19626350.
  24. Moodley KL, Owen CP, Patel M. Quantitative Analysis of Selected Microorganisms Present at Various Sites in a Prosthetics Clinic and Dental Laboratory during Complete Denture Fabrication. Int J Environ Res Public Health. 2020 May 12;17(10):3345. doi: 10.3390/ijerph17103345. PMID: 32408512; PMCID: PMC7277632.
  25. Delgado AHS, Carvalho J, Borrecho G, Nascimento T, Silva ME, Félix SA, Mendes JJ. In situ Multispecies Colonization of An Acrylic Resin: Comparison to Oral Microbiome and Potential for Inflammatory Response. Contemp Clin Dent. 2018 Jul-Sep;9(3):400-405. doi: 10.4103/ccd.ccd_141_18. PMID: 30166834; PMCID: PMC6104353.
  26. Ao A, Wakabayashi N, Nitta H, Igarashi Y. Clinical and microbiologic effects of lingual cervical coverage by removable partial dentures. Int J Prosthodont. 2013 Jan-Feb;26(1):45-50. doi: 10.11607/ijp.3061. PMID: 23342333.

Download Article
Received December 28, 2022.
Accepted February 18, 2023.
©2023 International Medical Research and Development Corporation.