Jsmc-10253

MOLECULAR IDENTIFICATION OF ENTAMOEBA HISTOLYTICA IN SULAIMANI PEDIATRIC TEACHING HOSPITAL

Sham Jamil Abdullah a, Shahnaz AbdulKadir Ali a, and Rizgar Raheem Sulaiman b

a Department of Microbiology, College of Medicine, University of Sulaimani, Kurdistan Region, Iraq.  

b Department of Medicine, College of Veterinary Medicine, University of Sulaimani, Kurdistan Region, Iraq. 

Submitted: 23/3/2019; Accepted: 27/7/2020; Published: 21/9/2020

DOI Link: https://doi.org/10.17656/jsmc.10253 

ABSTRACT

Background 

Entamoeba histolytica is a unicellular protozoon that conceder as a common cause of dysentery in human especially in the children under 6 years old. This may be transmitted via the ingestion of cystic stage of the parasite. 

Objectives 

This study aimed to investigate the total rate of Entamoeba histolytica infection in children admitted to the Pediatric Teaching Hospital in Sulaimani City, and considering some other aspects such as culturing and detecting subtypes using polymerase chain reaction (PCR). 

Patients and Methods

This epidemiologic study includes 1005 stool samples collected from children aged 6 months to 6 years old from the 1st of Jun to the 30th of October 2014. Each stool sample was examined and identified by direct wet mount method using normal physiological saline and iodine solution, followed by culturing and isolation of the parasite for 14 days to elongate life span and then genomic amplification applied from the cultured Entamoeba histolytica by appropriate PCR based method.

Results

The result of rapid kit occult blood combined with microscopic wet mount (iodine) showed that the infection with E. histolytica was (12.9%), in which the rate in male (14.18%) was higher than that female (11.42%), with no significant difference between both genders. cultured E. histolytica showed a great ability of this parasite to grow, with detection of the parasite under light microscope (40X). Then two standard kits were used specific primer (1147, F&R), and thus about 1147bp PCR produced. Then, nested PCR was performed for the same PCR products two different primers (246 F&R) which resulted in fragments of approximately 246bp. This fragment coding for the 16S SSUrDNA gene of HM-1IMSS 16s E. histolytica confirms the previous PCR amplification finding. The results of sequence methods identified E. histolytica subtype 1 from PCR positive samples by sequencing alignment (99%) similarity with accession number KB823016. 

Conclusion

The prevalence rate of infection with E. histolytica in Pediatric Teaching Hospital in Sulaimani City was (12.9%). Beef extract medium for culturing of E. histolytica showed a great ability for growing of the parasite cystic form. E. histolytica subtype1 can be detected by sequencing method for the first time in molecular genotype of human E .histolytica in Sulaimani city.

KEYWORDS

Molecular; Survey scale; Entamoeba histolytica.

References 

1. Stanley SL, Jr. Amoebiasis. Lancet (London, England). 2003 Mar 22;361(9362):1025-34. PubMed PMID: 12660071. Epub 2003/03/28. eng.

2. Molan AL, M FA. Prevalency of intestinal parasite in school children of arbil, north of Iraq. med J. 1989;10:107-10.

3. A A-SA, K HM. Parastic infection among primary school children in the tikreet city. SciJTikreet unvi. 1995;1(1):71-4.

4. Abdel-husaen, A JA. ntestinal parasit. journal of Ibn-Al-Haitham. 1994 2(2):1-18.

5. Rhadi HA. A survey of Intestinal Pathoggenic Parasite In Basrah City. echResJ. 1994;7(20):56-62.

6. Al-Dawdi and Khudaer. A.A. Poipiscafiwom, .MSc thesis. Prevalency of intestinal parasite in school children and food industrial workers of muesl [MSc]: Musel north of Iraq; 1998.

7. Al-Kubaise and Maki. A. H. (2000). study on immunity of patient infected with tissue amoebaesis [PhD. Thesis]. Baghdad biolo.Unvi. Al-Mustansrya.; 2000.

8. F.A. A-GaN. tudy on intestinal parasite in baghdad city [MSc thesis]. Baghdad Biolo.Univer.Al-Mustansrya; 2002.

9. Ali SA, Mohammed LO. Prevalence of intestinal parasite among children in Sulaimani City. JDuhokUniv. 2010;13(1):94-8.

10. Tengku SA, Norhayati M. Public health and clinical importance of amoebiasis in Malaysia: a review. Tropical biomedicine. 2011 Aug;28(2):194-222. PubMed PMID: 22041740. Epub 2011/11/02. eng.

11. Jackson TF, Suparsad S. Zymodeme stability of Entamoeba histolytica and E. dispar. Archives of medical research. 1997;28 Spec No:304-5. PubMed PMID: 9033111. Epub 1997/01/01. eng.

12. Cox FEG. History of human parasitology. Clin Microbiol Rev. 2002;15(4):595-612. PubMed PMID: 12364371. eng.

13. Calderaro A, Gorrini C, Bommezzadri S, Piccolo G, Dettori G, Chezzi C. Entamoeba histolytica and Entamoeba dispar: comparison of two PCR assays for diagnosis in a non-endemic setting. Transactions of the Royal Society of Tropical Medicine and Hygiene. 2006 May;100(5):450-7. PubMed PMID: 16274714. Epub 2005/11/09. eng.

14. Zaki M, Meelu P, Sun W, Clark CG. Simultaneous differentiation and typing of Entamoeba histolytica and Entamoeba dispar. J Clin Microbiol. 2002;40(4):1271-6. PubMed PMID: 11923344. eng.

15. Troll H, Marti H, Weiss N. Simple differential detection of Entamoeba histolytica and Entamoeba dispar in fresh stool specimens by sodium acetate-acetic acid-formalin concentration and PCR. J Clin Microbiol. 1997;35(7):1701-5. PubMed PMID: 9196177. eng.

16. Evangelopoulos A, Spanakos G, Patsoula E, Vakalis N, Legakis N. A nested, multiplex, PCR assay for the simultaneous detection and differentiation of Entamoeba histolytica and Entamoeba dispar in faeces. Annals of tropical medicine and parasitology. 2000 Apr;94(3):233-40. PubMed PMID: 10884867. Epub 2000/07/08. eng.

17. Katzwinkel-Wladarsch S, Loscher T, Rinder H. Direct amplification and differentiation of pathogenic and nonpathogenic Entamoeba histolytica DNA from stool specimens. The American journal of tropical medicine and hygiene. 1994 Jul;51(1):115-8. PubMed PMID: 8059910. Epub 1994/07/01. eng.

Full Text

 © The Authors, published by University of Sulaimani, College of Medicine

This work is licensed under a Creative Commons Attribution 4.0 International License.