Molecular Identification of Entamoeba gingivalis and Its Association with Periodontitis in Diabetic Patients: A PCR-Based Study Targeting the ITS2 Region

ORIGINAL ARTICLE
Wirangrong Sritongklang, Nav La, Alisa Boonsuya, Patpicha Arunsan, Thanawat Trasaktaweesakul, Pundit Asavaritikrai, Nathkapach Kaewpitoon Rattanapitoon, Schawanya Kaewpitoon Rattanapitoon

 
For citation: Sritongklang W, La N, Boonsuya A, Arunsan P, Trasaktaweesakul T, Asavaritikrai P, Rattanapitoon NK, Rattanapitoon SK. Molecular Identification of Entamoeba gingivalis and Its Association with Periodontitis in Diabetic Patients: A PCR-Based Study Targeting the ITS2 Region. International Journal of Biomedicine. 2025;15(3):545-551. doi:10.21103/Article15(3)_OA15
 
Originally published September 5, 2025

Abstract: 

Entamoeba gingivalis is an anaerobic protozoan increasingly associated with periodontitis, particularly in individuals with systemic conditions such as diabetes mellitus. This cross-sectional study investigated the prevalence of E. gingivalis in 160 Thai patients with periodontitis and analyzed associated risk factors. Microscopy detected the parasite in 15.0% of patients, PCR in 25.6%, and sequencing confirmed the infection in 22.5%, all of which were Subtype 1. Infection was significantly more common among diabetic patients (41.3%) than non-diabetics (3.8%) (P<0.001). Multivariate analysis identified type 2 diabetes (adjusted OR = 20.77; 95% CI: 5.39–80.09), alcohol use (adjusted OR = 3.72; 95% CI: 1.01–13.68), and other underlying diseases (adjusted OR = 1.49; 95% CI: 1.07–2.06) as independent risk factors. PCR demonstrated superior diagnostic performance compared to microscopy. These findings support a potential pathogenic role of E. gingivalis in periodontitis, especially among patients with type 2 diabetes or behavioral risk factors. 

Keywords: 
Entamoeba gingivalis • periodontitis • diabetes mellitus • internal transcribed spacer 2 • polymerase chain reaction
References: 
  1. Bao X, Wiehe R, Dommisch H, Schaefer AS. Entamoeba gingivalis Causes Oral Inflammation and Tissue Destruction. J Dent Res. 2020 May;99(5):561-567. doi: 10.1177/0022034520901738. Epub 2020 Feb 5. PMID: 32023135.
  2. Bartold PM, Van Dyke TE. An appraisal of the role of specific bacteria in the initial pathogenesis of periodontitis. J Clin Periodontol. 2019 Jan;46(1):6-11. doi: 10.1111/jcpe.13046. PMID: 30556922; PMCID: PMC6357965.
  3. Bhaijee F. Entamoeba gingivalis: The oral amoeba—A review. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2011;112(4):406–411. doi: 10.1016/j.tripleo.2011.05.016
  4. Bonner M, Fresno M, Gironès N, Guillén N, Santi-Rocca J. Reassessing the Role of Entamoeba gingivalis in Periodontitis. Front Cell Infect Microbiol. 2018 Oct 29;8:379. doi: 10.3389/fcimb.2018.00379. PMID: 30420943; PMCID: PMC6215854.
  5. Jiao J, Bie M, Xu X, Duan D, Li Y, Wu Y, Zhao L. Entamoeba gingivalis is associated with periodontal conditions in Chinese young patients: A cross-sectional study. Front Cell Infect Microbiol. 2022 Oct 7;12:1020730. doi: 10.3389/fcimb.2022.1020730. PMID: 36275028; PMCID: PMC9585380.
  6. Boonsuya A, Chitpitaklert P, Pechdee P, Srithongklang W, Thanchonnang C, La N, Gordon CN, Rattanapitoon NK, Arunsan P, Rattanapitoon SK. Oral parasitic protozoan Entamoeba gingivalis in periodontal disease patients, northeastern Thailand. Trop Biomed. 2023 Dec 1;40(4):471-477. doi: 10.47665/tb.40.4.013. PMID: 38308835.
  7. Papapanou PN, Sanz M, Buduneli N, Dietrich T, Feres M, Fine DH, Flemmig TF, Garcia R, Giannobile WV, Graziani F, Greenwell H, Herrera D, Kao RT, Kebschull M, Kinane DF, Kirkwood KL, Kocher T, Kornman KS, Kumar PS, Loos BG, Machtei E, Meng H, Mombelli A, Needleman I, Offenbacher S, Seymour GJ, Teles R, Tonetti MS. Periodontitis: Consensus report of workgroup 2 of the 2017 World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions. J Periodontol. 2018 Jun;89 Suppl 1:S173-S182. doi: 10.1002/JPER.17-0721. PMID: 29926951.
  8. GOMORI G. A rapid one-step trichrome stain. Am J Clin Pathol. 1950 Jul;20(7):661-4. doi: 10.1093/ajcp/20.7_ts.661. PMID: 15432364.
  9. Khan M, Alshahrani S, Alqhtani A, Alzahrani A, Farouk AE. Molecular detection of Entamoeba gingivalis in diabetic patients with periodontitis: A case-control study. Saudi J Biol Sci. 2020;27(3):920–925. doi: 10.1016/j.sjbs.2020.01.001
  10. Garcia G, Ramos F, Maldonado J, Fernandez A, Yáñez J, Hernandez L, Gaytán P. Prevalence of two Entamoeba gingivalis ST1 and ST2-kamaktli subtypes in the human oral cavity under various conditions. Parasitol Res. 2018 Sep;117(9):2941-2948. doi: 10.1007/s00436-018-5990-8. Epub 2018 Jul 9. PMID: 29987412.
  11. Graziani F, Cei S, Orlandi M, Gabriele M, Gennai S, Filice N, et al. Hypertension and periodontitis: The results of the periodontal disease and Vascular Events (PAVE) study. J Clin Periodontol. 2018;45(6):789–798. doi: 10.1111/jcpe.12827
  12. Mauri-Obradors E, Estrugo-Devesa A, Jane-Salas E, Vinas M, Lopez-Lopez J. Impact of dyslipidemia on the periodontium: An umbrella review. J Evid Based Dent Pract. 2024;24(1):101842. doi: 10.1016/j.jebdp.2023.101842
  13. Costa FO, da Rocha ACB, da Silva JFF, Mendonça JA, Costa JEM, Batista MJ, et al. High prevalence of Entamoeba gingivalis in HIV-infected individuals with periodontal disease. Acta Trop. 2022;232:106515. doi: 10.1016/j.actatropica.2022.106515
  14. Nazir MA. Prevalence of periodontal disease, its association with systemic diseases and prevention. Int J Health Sci (Qassim). 2017 Apr-Jun;11(2):72-80. PMID: 28539867; PMCID: PMC5426403.
  15. Vlasa A, Bud A, Lazar L, Lazar AP, Herbert A, Bud E. Association of Entamoeba gingivalis with Periodontal Disease-Systematic Review and Meta-Analysis. Medicina (Kaunas). 2024 Apr 29;60(5):736. doi: 10.3390/medicina60050736. PMID: 38792919; PMCID: PMC11123156.
  16. Bonner M, Amard V, Bar-Pinatel C, Charpentier F, Chatard JM, Desmuyck Y, Ihler S, Rochet JP, Roux de La Tribouille V, Saladin L, Verdy M, Gironès N, Fresno M, Santi-Rocca J. Detection of the amoeba Entamoeba gingivalis in periodontal pockets. Parasite. 2014;21:30. doi: 10.1051/parasite/2014029. Epub 2014 Jul 2. PMID: 24983705; PMCID: PMC4077299.
  17. Jalallou N, Rostami-Nejad M, Maleki D, Mirjalali H, Zali MR. Molecular detection and genotyping of Entamoeba gingivalis in patients with periodontal disease in Iran. Parasitol Int. 2019;72:101922. doi: 10.1016/j.parint.2019.101922
  18. El Deeb HK, Allam AA, Ahmed A, Attia RA, Gad El-Rab MO, Kandil OM, et al. Genotyping of Entamoeba gingivalis in periodontitis patients: Evidence of subtype 1 predominance. Acta Trop. 2020;205:105404. doi: 10.1016/j.actatropica.2020.105404

Download Article
Received June 16, 2025.
Accepted July 31, 2025.
©2025 International Medical Research and Development Corporation.