DIENTAMOEBA FRAGILIS AND GASTROINTESTINAL SYMPTOMS: EFFICACY OF DIAGNOSTIC METHODS AND GENOTYPE-SYMPTOM CORRELATION

Abstract

Objective: This study aimed to investigate the association between Dientamoeba fragilis and gastrointestinal symptoms, assess the effectiveness of various diagnostic methods, and determine the correlation between genotypes and clinical manifestations. Material and Methods: Stool samples were collected from 116 patients with gastrointestinal complaints. The presence of D. frag ilis was investigated using culture methods (BD and LS media) and molecular techniques (real-time PCR and conventional PCR). Positive samples were subjected to sequencing analysis for geno typing. Results: D. fragilis was detected in 18 out of 116 patients (15.52%) using PCR, whereas culture methods identified only 9 positive cases (7.75%). A statistically significant difference was observed between the molecular and culture methods (p<0.001). All D. frag ilis samples belonged to Genotype 1. The most common symptoms were diarrhoea (76.9%) and abdominal pain (61.5%); however, no significant correlation was found between genotype and clinical symptoms. Conclusion: Real-time PCR was the most sensitive and specific diagnostic method. Genotyping results indicated no association between D. fragilis genotypes and clinical manifestations. Further studies are needed to explore additional genomic targets to better understand the pathogenic potential of D. fragilis.

Keywords

• Dientamoeba fragilis
• culture
• genotyping

DIENTAMOEBA FRAGILIS VE GASTROİNTESTİNAL SEMPTOMLAR: TANI YÖNTEMLERİNİN ETKİNLİĞİ VE GENOTİP-BULGU İLİŞKİSİ

Abstract

Amaç: Bu çalışma, Dientamoeba fragilis ile gastrointestinal semp tomlar arasındaki ilişkiyi araştırmayı, farklı tanı yöntemlerinin etkinliğini değerlendirmeyi ve genotipler ile klinik bulgular arasın daki korelasyonu belirlemeyi amaçlamaktadır. Gereç ve Yöntemler: Gastrointestinal şikayetleri olan 116 hastadan dışkı örnekleri toplandı. D. fragilis varlığı kültür yöntemleri (BD ve LS besiyerleri) ve moleküler teknikler (real-time PCR ve konvan siyonel PCR) kullanılarak araştırıldı. Pozitif örnekler genotipleme amacıyla sekans analizine tabi tutuldu. Bulgular: PCR ile 116 hastanın 18’inde (%15,52) D. fragilis saptandı. Buna karşın, kültür yöntemleri yalnızca 9 pozitif vaka (%7,75) belirledi. Moleküler yöntemler ile kültür yöntemleri arasında ista tistiksel olarak anlamlı bir fark bulundu (p<0,001). Tüm D. fragilis örnekleri Genotip 1 olarak tanımlandı. En yaygın semptomlar ishal (%76,9) ve karın ağrısı (%61,5) idi; ancak, genotip ile klinik semp tomlar arasında anlamlı bir ilişki saptanmadı. Sonuç: Real-time PCR, en duyarlı ve spesifik tanı yöntemi olarak belirlendi. Genotipleme sonuçları, D. fragilis genotipleri ile klinik semptomlar arasında bir ilişki bulunmadığını göstermiştir. D. fragilis’in patojenite potansiyelini daha iyi anlamak için ek genomik hedeflerin araştırılmasına yönelik ileri çalışmalara ihtiyaç vardır.

Keywords

• Dientamoeba fragilis
• kültür
• genotiplendirme

References

1. Stark D, Barratt J, Chan D, Ellis JT. Dientamoeba fragilis, the neglected trichomonad of the human bowel. Clin Microbiol Rev 2016;29(3):553-80. google scholar
2. Jepps MW, Dobell C. Dientamoeba fragilis, a new intestinal amoeba from man. Parasitology 1918;10(3):352-67. google scholar
3. Garcia LS. Dientamoeba fragilis, one of the neglected intestinal protozoa. J Clin Microbiol 2016;54(9):2243-50. google scholar
4. Stark D, Garcia LS, Barratt JL, Phillips O, Roberts T, Marriott D, et al. Description of Dientamoeba fragilis cyst and precystic forms from human samples. J Clin Microbiol 2014;52(7):2680-3. google scholar
5. Cacciò SM. Molecular epidemiology of Dientamoeba fragilis. Acta Trop 2018;184:73-7. google scholar
6. Stark D, Barratt J, Roberts T, Marriott D, Harkness J, Ellis J. A review of the clinical presentation of dientamoebiasis. Am J Trop Med Hyg 2010;82(4):614-9. google scholar
7. Munasinghe VS, Vella NGF, Ellis JT, Windsor PA, Stark D. Cyst formation and faecal–oral transmission of Dientamoeba fragilis – the missing link in the life cycle of an emerging pathogen. International Journal for Parasitology 2013;43(11):879-83. google scholar
8. Norberg A, Nord CE, Evengård B. Dientamoeba fragilis-a protozoal infection which may cause severe bowel distress. Clin Microbiol Infect 2003;9(1):65-8. google scholar

9. Johnson EH, Windsor JJ, Clark CG. Emerging from obscurity: biological, clinical, and diagnostic aspects of Dientamoeba fragilis. Clin Microbiol Rev 2004;17(3):553-70. google scholar
10. Clark CG, Diamond LS. Methods for cultivation of luminal parasitic protists of clinical importance. Clin Microbiol Rev 2002;15(3):329-41. google scholar
11. Barratt JL, Banik GR, Harkness J, Marriott D, Ellis JT, Stark D. Newly defined conditions for the in vitro cultivation and cryopreservation of Dientamoeba fragilis: new techniques set to fast track molecular studies on this organism. Parasitology 2010;137(13):1867-78. google scholar
12. Munasinghe VS, Stark D, Ellis JT. New advances in the in-vitro culture of Dientamoeba fragilis. Parasitology 2012;139(7):864-9. google scholar
13. Peek R, Reedeker FR, van Gool T. Direct amplification and genotyping of Dientamoeba fragilis from human stool specimens. J Clin Microbiol 2004;42(2):631-5. google scholar
14. Miguel L, Salvador F, Sulleiro E, Sánchez-Montalvá A, Molina-Morant D, López I, et al. Clinical and epidemiological characteristics of patients with Dientamoeba fragilis infection. Am J Trop Med Hyg 2018;99(5):1170-3. google scholar
15. Barratt JL, Harkness J, Marriott D, Ellis JT, Stark D. A review of Dientamoeba fragilis carriage in humans: several reasons why this organism should be considered in the diagnosis of gastrointestinal illness. Gut Microbes 2011;2(1):3-12. google scholar
16. Yakoob J, Jafri W, Beg MA, Abbas Z, Naz S, Islam M, et al. Blastocystis hominis and Dientamoeba fragilis in patients fulfilling irritable bowel syndrome criteria. Parasitol Res 2010;107(3):679-84. google scholar
17. Jirků M, Kašparová A, Lhotská Z, Oborník M, Brožová K, Petrželková KJ, et al. A cross-sectional study on the occurrence of the intestinal protist, Dientamoeba fragilis, in the gut-healthy volunteers and their animals. Int J Mol Sci 2022;23(23):15407. google scholar
18. Stark D, Beebe N, Marriott D, Ellis J, Harkness J. Evaluation of three diagnostic methods, including real-time PCR, for detection of Dientamoeba fragilis in stool specimens. J Clin Microbiol 2006;44(1):232-5. google scholar
19. Stensvold CR, Clark CG, Röser D. Limited intra-genetic diversity in Dientamoeba fragilis housekeeping genes. Infect Genet Evol 2013;18:284-6. google scholar
20. Cacciò SM, Sannella AR, Bruno A, Stensvold CR, David EB, Guimarães S, et al. Multilocus sequence typing of Dientamoeba fragilis identified a major clone with widespread geographical distribution. Int J Parasitol 2016;46(12):793-8. google scholar
21. Stark DJ, Beebe N, Marriott D, Ellis JT, Harkness J. Dientamoebiasis: clinical importance and recent advances. Trends Parasitol 2006;22(2):92-6. google scholar
22. Stark D, Barratt J, Roberts T, Marriott D, Harkness J, Ellis J. Comparison of microscopy, two xenic culture techniques, conventional and real-time PCR for the detection of Dientamoeba fragilis in clinical stool samples. Eur J Clin Microbiol Infect Dis 2010;29(4):411-6. google scholar
23. Yang J, Scholten T. Dientamoeba fragilis: a review with notes on its epidemiology, pathogenicity, mode of transmission, and diagnosis. Am J Trop Med Hyg 1977;26(1):16-22. google scholar
24. Lagacé-Wiens PR, VanCaeseele PG, Koschik C. Dientamoeba fragilis: an emerging role in intestinal disease. CMAJ 2006;175(5):468-. google scholar
25. Banik GR, Barratt JL, Marriott D, Harkness J, Ellis JT, Stark D. A case-controlled study of Dientamoeba fragilis infections in children. Parasitology 2011;138(7):819-23. google scholar
26. Ögren J, Dienus O, Löfgren S, Einemo IM, Iveroth P, Matussek A. Dientamoeba fragilis prevalence coincides with gastrointestinal symptoms in children less than 11 years old in Sweden. Eur J Clin Microbiol Infect Dis 2015;34(10):1995-8. google scholar
27. Spencer MJ, Garcia LS, Chapin MR. Dientamoeba fragilis. An intestinal pathogen in children? Am J Dis Child 1979;133(4):390-3. google scholar
28. Stark D, Beebe N, Marriott D, Ellis J, Harkness J. Prospective study of the prevalence, genotyping, and clinical relevance of Dientamoeba fragilis infections in an Australian population. J Clin Microbiol 2005;43(6):2718-23. google scholar
29. Preiss U, Ockert G, Brömme S, Otto A. Dientamoeba fragilis infection, a cause of gastrointestinal symptoms in childhood. Klinische Pädiatrie 1990;202(02):120-3. google scholar
30. Clemente L, Pasut M, Carlet R, Ruscio M, Fontana F. Dientamoeba fragilis in the North-East of Italy: Prevalence study and treatment. Parasitol Int 2021;80:102227. google scholar
31. Vandenberg O, Peek R, Souayah H, Dediste A, Buset M, Scheen R, et al. Clinical and microbiological features of dientamoebiasis in patients suspected of suffering from a parasitic gastrointestinal illness: A comparison of Dientamoeba fragilis and Giardia lamblia infections. Int J Infect Dis 2006;10(3):255-61. google scholar
32. Vassalou E, Vassalos CM, Spanakos G, Fotopoulos A, Dounias G, Kalofolias P, et al. First report of Dientamoeba fragilis infection explaining acute non-specific abdominal pain. Indian J Med Microbiol 2016;34(1):106-8. google scholar
33. Pietilä JP, Meri T, Siikamäki H, Tyyni E, Kerttula AM, Pakarinen L, et al. Dientamoeba fragilis - the most common intestinal protozoan in the Helsinki Metropolitan Area, Finland, 2007 to 2017. Euro Surveill 2019;24(29):1800546. google scholar
34. Brands MR, Van de Vijver E, Haisma SM, Heida A, van Rheenen PF. No association between abdominal pain and Dientamoeba in Dutch and Belgian children. Arch Dis Child 2019;104(7):686-9. google scholar
35. Cuffari C, Oligny L, Seidman EG. Dientamoeba fragilis masquerading as allergic colitis. J Pediatr Gastroenterol Nutr 1998;26(1):16-20. google scholar
36. Shein R, Gelb A. Colitis due to Dientamoeba fragilis. Am J Gastroenterol 1983;78(10):634-6. google scholar
37. Unalan-Altintop T, Vahabov C, Ergunay K, Kurt O, Kav T, Akyon Y, et al. Investigation of Dientamoeba fragilis and Blastocystis in patients from Turkey with ulcerative colitis and irritable bowel syndrome: Any relation with genotypes? Acta Trop 2022;231:106451. google scholar
38. Petersen AM, Stensvold CR, Mirsepasi H, Engberg J, Friis-Møller A, Porsbo LJ, et al. Active ulcerative colitis associated with low prevalence of Blastocystis and Dientamoeba fragilis infection. Scand J Gastroenterol 2013;48(5):638-9. google scholar
39. Rostami A, Riahi SM, Haghighi A, Saber V, Armon B, Seyyedtabaei SJ. The role of Blastocystis sp. and Dientamoeba fragilis in irritable bowel syndrome: a systematic review and meta-analysis. Parasitol Res 2017;116(9):2361-71. google scholar
40. Vezir S, Kaya F, Vezir E, Karaosmanoğlu N, Adiloğlu AK. Evaluation of intestinal parasites in patients with chronic spontaneous urticaria in a territory hospital in Turkey. J Infect Dev Ctries 2019;13(10):927-32. google scholar
41. Sarzhanov F, Dogruman-Al F, Santin M, Maloney JG, Gureser AS, Karasartova D, et al. Investigation of neglected protists Blastocystis sp. and Dientamoeba fragilis in immunocompetent and immunodeficient diarrheal patients using both conventional and molecular methods. PLoS Negl Trop Dis 2021;15(10):e0009779. google scholar
42. Hawash YA, Ismail KA, Saber T, Eed EM, Khalifa AS, Alsharif KF, et al. Dientamoeba fragilis infection in patients with digestive and non-digestive symptoms: a case-control study. Korean J Parasitol 2020;58(2):129-34. google scholar
43. Sarafraz S, Farajnia S, Jamali J, Khodabakhsh F, Khanipour F. Detection of Dientamoeba fragilis among diarrheal patients referred to Tabriz health care centers by nested PCR. Trop Biomed 2013;30(1):113-8. google scholar
44. Yıldız İ, Ertuğ S, Tileklioğlu E, Malatyalı E, Güçlü Ö, Ertabaklar H. Investigation of Dientamoeba fragilis frequency in faecal samples of patients with Enterobius vermicularis infection by polymerase chain reaction. Turkiye Parazitol Derg 2021;45(3):195-200. google scholar
45. Girginkardeşler N, Kurt O, Kilimcioğlu AA, Ulgen Ok Z. Transmission of Dientamoeba fragilis: Evaluation of the role of Enterobius vermicularis. Parasitol Int 2008;57(1):72-5. google scholar
46. Menéndez C, Fernández-Suarez J, Boga Ribeiro JA, Rodríguez-Pérez M, Vázquez F, Gonzalez-Sotorrios N, et al. Epidemiological and clinical characteristics of Dientamoeba fragilis infection. Enferm Infecc Microbiol Clin (Engl Ed) 2019;37(5):290-5. google scholar