year 18, Issue 3 (May - June 2024)                   Iran J Med Microbiol 2024, 18(3): 172-180 | Back to browse issues page


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Karem Kadhim M, Majid Khazaal R, Hassan Flaih M, Reissan Hussein K. Molecular Characterization and Phylogenetic Analysis of the ITS2 Gene of Enterobius vermicularis Isolates. Iran J Med Microbiol 2024; 18 (3) :172-180
URL: http://ijmm.ir/article-1-2069-en.html
1- Thi-Qar Education Directorate, Nasiriyah, Iraq
2- Department of Medical Laboratory Techniques, Nasiriyah Technical Institute, Southern Technical University, Nasiriyah, Iraq , ruaa.m1982@stu.edu.iq
3- Department of Nursing Techniques, Nasiriyah Technical Institute, Southern Technical University, Nasiriyah, Iraq
Abstract:   (291 Views)

Background and Aim: Enterobius vermicularis (E. vermicularis) is a parasitic nematode infecting humans, mainly children, with a prevalence rate above 35% in some countries. Its molecular studies are still scant. Precise diagnosis and understanding of genetic variation are essential in managing enterobiasis. This study used DNA sequencing and phylogenetic analysis to detect E. vermicularis by genetic variation in the internal transcribed spacer 2 (ITS2) gene.
Materials and Methods: Stool samples were collected randomly from 56 infected patients and 60 healthy individuals as the control group. Genomic DNA was isolated, and PCR was done using primers against ITS2 region of rDNA. Our research was conducted with the utmost thoroughness, as evidenced by DNA sequencing determining the genetic associations and variations of rDNA (ITS2) among E. vermicularis isolates. The sequences were then compared with NCBI-BLAST sequences for the genetic homology and variation analysis, ensuring the accuracy and reliability of the results.
Results: In the molecular testing, 32 of the 56 samples were positive for the ITS2 gene of E. vermicularis. The percentage identity of local E. vermicularis IQN.1-10 isolates with those available in the NCBI-BLAST database homology ranged from 99.54% to 99.56%. The genetic variation analysis revealed three deletions and substitution mutations, with a total variation percentage of 0.44% to 0.46%. These findings not only provide a deeper understanding of E. vermicularis but also hold promise for the development of more effective strategies for managing enterobiasis, giving hope for the future.
Conclusion: The results indicated high accuracy of the molecular diagnosis and the importance of ITS2 as the most important genetic marker due to its highly conserved nature. These new data on E. vermicularis genotypes from Iraq reflect the diversity and thus warrant further molecular studies across Iraq to effectively consider the genetic landscape and help in diagnostic and control measures.

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Type of Study: Original Research Article | Subject: Microbial Genetics
Received: 2024/04/27 | Accepted: 2024/08/1 | ePublished: 2024/08/18

References
1. Fantinatti M, Da-Cruz AM. Enterobius vermicularis in Brazil: An integrative review. Revista da Soc Brasil de Med Trop. 2023;56:e0073-2023. [DOI:10.1590/0037-8682-0073-2023] [PMID] [PMCID]
2. Moussavi E, Houssaini M, Salari N, Hemmati M, Abdullahi A, Khaleghi AA, et al. Prevalence of Enterobius vermicularis among children in Iran: A comprehensive systematic review and meta-analysis. Parasite Epidemiol Control. 2023;22:e00315. [DOI:10.1016/j.parepi.2023.e00315] [PMID] [PMCID]
3. Yildiz I, Malatyali E, Tilekliogu E, Ertabaklar H, Ertug S. A retrospective analysis of Enterobius vermicularis frequency for the last five years in Aydin, Turkey. Ann Med Res. 2021;28(9):1716-9. [DOI:10.5455/annalsmedres.2020.10.1053]
4. Actor JK. Adaptive Immune Response and Hypersensitivity. In Elsevier's Integrated Review Immunology and Microbiology. 2nd ed. pp.53-9. 2012. Philadelphia, U.S.A.: Elsevier. [DOI:10.1016/B978-0-323-07447-6.00007-7]
5. Tavan A, Mikaeili F, Sadjjadi SM, Bajelan S, Mahmoudvand H, Sharifdini M. Prevalence and genotype distribution of Enterobius vermicularis among kindergarteners in Shiraz and Khorramabad cities, Iran. Asian Pac J Trop Med. 2020;13(7):308-13. [DOI:10.4103/1995-7645.280229]
6. Fan CK, Sonko P, Lee YL, Yin AW, Chuang TW, Kios R, et al. Epidemiologic Study of Enterobius vermicularis Infection among Schoolchildren in the Republic of Marshall Islands. J Trop Med. 2021;2021:6273954. [DOI:10.1155/2021/6273954] [PMID] [PMCID]
7. Wendt S, Trawinski H, Schubert S, Rodloff AC, Mössner J, Lübbert C. The Diagnosis and Treatment of Pinworm Infection. Dtsch Arztebl Int. 2019;116(13):213-9. [DOI:10.3238/arztebl.2019.0213] [PMID] [PMCID]
8. Yahya NA, Hamad SS, Muhammad DA. Isolation and Molecular Identification of Enterobius Vermicularis from Patient in Emigrant Campus. HIV Nursing. 2023;23(1):779-82.
9. Ummarino A, Caputo M, Tucci FA, Pezzicoli G, Piepoli A, Gentile A, et al. A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application. Front Microbiol. 2022;13:1028988. [DOI:10.3389/fmicb.2022.1028988] [PMID] [PMCID]
10. Abu Al-Soud W, Rådström P. Effects of amplification facilitators on diagnostic PCR in the presence of blood, feces, and meat. J Clin Microbiol. 2000;38(12):4463-70. [DOI:10.1128/JCM.38.12.4463-4470.2000] [PMID] [PMCID]
11. Morin PA, Chambers KE, Boesch C, Vigilant L. Quantitative polymerase chain reaction analysis of DNA from noninvasive samples for accurate microsatellite genotyping of wild chimpanzees (Pan troglodytes verus). Mol Ecol. 2001;10(7):1835-44. [DOI:10.1046/j.0962-1083.2001.01308.x] [PMID]
12. Eggert LS, Maldonado JE, Fleischer RC. Nucleic acid isolation from ecological samples--animal scat and other associated materials. Methods Enzymol. 2005;395:73-87. [DOI:10.1016/S0076-6879(05)95006-4] [PMID]
13. Rahnamaye V, Hagh H, Oskouei MM. Genetic Classification and Differentiation of Enterobius vermicularis Based on Mitochondrial Cytochrome C Oxidase (cox1) in Northwest of Iran. J Pure Appl Microbiol. 2014;8(5):3995-9.
14. Iñiguez AM, Vicente AC, Araújo A, Ferreira LF, Reinhard KJ. Enterobius vermicularis: specific detection by amplification of an internal region of 5S ribosomal RNA intergenic spacer and trans-splicing leader RNA analysis. E. vermicularis: specific detection by PCR and SL1 RNA analysis. Exp Parasitol. 2002;102(3-4):218-22. [DOI:10.1016/S0014-4894(03)00059-6] [PMID]
15. Qiu JH, Lou Y, Zhang Y, Chang QC, Liu ZX, Duan H, et al. Sequence variability in internal transcribed spacers of nuclear ribosomal DNA among isolates of the oxyurid nematodes Syphacia obvelata and Aspiculuris tetraptera from mice reared in laboratories in China. J Helminthol. 2016;90(1):81-5. [DOI:10.1017/S0022149X1400087X] [PMID]
16. Leblanc M, Berry K, Graciano S, Becker B, Reuter JD. False-positive results after environmental pinworm PCR testing due to Rhabditid nematodes in Corncob bedding. J Am Assoc Lab Anim Sci. 2014;53(6):717-24.
17. Schrader C, Schielke A, Ellerbroek L, Johne R. PCR inhibitors - occurrence, properties and removal. J Appl Microbiol. 2012;113(5):1014-26. [DOI:10.1111/j.1365-2672.2012.05384.x] [PMID]
18. Shilanabadi K, Derakhshan F, Raeghi S. Phylogenetic study of Enterobius vermicularis isolates from Northwest of Iran. Marag Univ Med Sci. 2022:1-9. [DOI:10.21203/rs.3.rs-1405714/v1]
19. Kaneva E, Tsvetkova N, Velcheva D, Harizanov R, Kaftandjiev I, Borisova R, et al. Development and Application of a Method for Genetic Detection of Enterobius Vermicularis in Samples of Patients With Enterobiasis. J IMAB. 2022;30(1):5295-300. [DOI:10.5272/jimab.2024301.5295]
20. Tomanakan K, Sanpool O, Chamavit P, Lulitanond V, Intapan PM, Maleewong W. Genetic variation of Enterobius vermicularis among schoolchildren in Thailand. J Helminthol. 2018;94:e7. [DOI:10.1017/S0022149X18000962] [PMID]
21. Zelck Ulrike E, Bialek R, Weiß M. Molecular Phylogenetic Analysis of Enterobius vermicularis and Development of an 18S Ribosomal DNA-Targeted Diagnostic PCR. J Clin Microbiol. 2020;49(4):1602-4. [DOI:10.1128/JCM.02454-10] [PMID] [PMCID]
22. Le TH, Blair D, McManus DP. Mitochondrial genomes of human helminths and their use as markers in population genetics and phylogeny. Acta Trop. 2000;77(3):243-56. [DOI:10.1016/S0001-706X(00)00157-1] [PMID]
23. Piperaki ET, Spanakos G, Patsantara G, Vassalou E, Vakalis N, Tsakris A. Characterization of Enterobius vermicularis in a human population, employing a molecular-based method from adhesive tape samples. Mol Cell Probes. 2011;25(2-3):121-5. [DOI:10.1016/j.mcp.2011.03.005] [PMID]
24. Shafiei R, Jafarzadeh F, Bozorgomid A, Ichikawa-Seki M, Mirahmadi H, Raeghi S. Molecular and phylogenetic analysis of E. vermicularis in appendectomy specimens from Iran. Infect Genet Evol. 2023;107:105391. [DOI:10.1016/j.meegid.2022.105391] [PMID]
25. Kaneva E, Harizanov R, Tsvetkova N, Kaftanjiev I, Borisova R, Ivanova A, et al. E. vermicularis – prospects for Future Research: A Brief Literature Review. Prob Infect Parasit Dis. 2022;50(2):13-8. [DOI:10.58395/pipd.v50i2.92]
26. Kubiak K, Dzika E. Enterobiasis epidemiology and molecular characterization of in healthy children in north-eastern Poland. Helminthologia. 2017;54(4):284-91. [DOI:10.1515/helm-2017-0042]
27. Al-Taei AHO, Al-Quraishi MA. Molecular identification of Enterobius vermicularis worms isolated from children in Babylon province of Iraq. Int J Health Sci. 2022;6(S6):9226-40. [DOI:10.53730/ijhs.v6nS6.12426]
28. Kadhum R, Jaber N, Alkhanaq M. Molecular identification and phylogenetic tree analysis of Enterobius vermicularis from Children in Wasit City of Iraq Address for correspondence. Indian J Sci. 2019;2018(8):14172-9.
29. Janthu P, Dumidae A, Subkrasae C, Ardpairin J, Nateeworanart S, Thanwisai A, Vitta A. Prevalence and genetic analysis of Enterobius vermicularis in schoolchildren in lower northern Thailand. Parasitol Res. 2022;121(10):2955-65. [DOI:10.1007/s00436-022-07626-0] [PMID]

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