Jsmc-10273

A COMPARATIVE STUDY OF GIARDIA LAMBLIA DETECTION BY USING OF MICROSCOPIC, SEROLOGIC, AND MOLECULAR METHODS, IN SULAIMANI PROVINCE

Safin Hamad Ismail a and Shahnaz AbdulKadir Ali b

a Ministry of Health, Sulaimani Directorate of Health, Public Health Laboratory, Kurdistan Region, Iraq. 

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

Submitted: 2/12/2019; Accepted: 1/12/2020; Published: 21/12/2020

DOI Link: https://doi.org/10.17656/jsmc.10273 

ABSTRACT

Background 

Giardia lamblia is an intestinal flagellated protozoan parasite that infects humans and animals. Giardiasis causing more than 200 million symptomatic infections globally it is one of the most common causes of diarrhea in developing countries, and frequent cause water-born/food-born parasitic diseases. 

Objectives

This study was carried out to investigate the prevalence rate of G. lamblia in Sulaimani Province hospitals and to identify the best and accurate method for identification. 

Patients and Methods

In the present study, 355 fecal specimens were collected from patients in three hospitals (Dr. Jamal A. Rashid Pediatric Teaching Hospitals, Ibrahim Pasha Health Center and Directorate of Preventive Health) in Sulaimani Province during the 1st November 2018 to the 30 April 2019. All samples examined by direct microscopic examination using Saline Wet Mount, and antigen detection by ELISA technique (RIADASCREEN Giardia ELISA kit). DNA extracted by used (QIAamp Fast DNA Stool Mini Kit 50), and for PCR amplification, used JW1, the forward primer: 5’ GCG CAC CAG GAATGT CTT GT 3’ and JW2, the reverse primer 5’ TCA CCT ACG GAT ACC TTG TT 3’ to amplify a 183-bp region of the 18S G. lamblia rRNA gene.

Results

Out of 355 stool samples, fifty samples were positive for G. lamblia at a prevalence rate of (14.1%) by Direct wet mount and ELISA, and forty of them were positive (11.4%) by PCR. Males showed a higher (18.1%) prevalence rate than females (5.4%). The highest rate (26.9%) of infection was found in (13-18) age groups, while the lowest rate (2.3%) was fond in (6-12) year of ages. The prevalence rate in rural area was higher than urban area (15.7%), (13.4%) respectively. The sequences alignments were 91.30% and 98.52% similar to M90523.1 and M90524.1 respectively. In comparison to Microscopic exam, both ELISA and PCR recorded the same specificity rate 99.67% as direct wet mount, but were different sensitivity rate 100% and 80% respectively. 

Conclusion

Direct wet mount and ELISA was more sensitive than PCR (80%), but they were specific at the same rate (99.67%), and this indicate that there is more than one gene of G. lamblia is endemic in Sulaimani Province.

KEYWORDS

Giardia; Direct wet mount; ELISA; PCR; Comparison.

References 

1. Rahimian F, Sadraei J, Pirestani M, Ghaffarifar F. A modified PCR-RFLP method to determine genetic diversity of Giardia lamblia human isolates based on triosephosphate isomerase (TPI) gene. Acta Trop. 2018;186:58-62.

2. Beyhan YE, Tas Cengiz Z. Comparison of microscopy, ELISA, and real-time PCR for detection of Giardia intestinalis in human stool specimens. Turk J Med Sci. 2017;47(4):1295-9.

3. Cernikova L, Faso C, Hehl AB. Five facts about Giardia lamblia. PLoS Pathog.2018;14(9):e1007250.

4. Lass A, Karanis P, Korzeniewski K. First detection and genotyping of Giardia intestinalis in stool samples collected from children in Ghazni Province, eastern Afghanistan and evaluation of the PCR assay in formalin-fixed specimens. Parasitology research. 2017;116(8):2255-64.

5. Gasparinho C, Ferreira FS, Mayer AC, Mirante MC, Vaz Nery S, Santos-Reis A, et al. Molecular characterization of Giardia lamblia in children less than 5 years of age with diarrhoea attending the Bengo General Hospital, Angola. Trans R Soc Trop Med Hyg.2017;111(11):497-503.

6. Mardu F, Berhe B, Tesfay K, Negash H. Assessment of sanitary condition of services 

 as implication for intestinal parasitic infections among prison inmates: institutional based cross-sectional study in eastern Tigrai zonal prison, northern Ethiopia, 2018. BMC Res Notes. 2019;12(1):393.

7. Pestechian N, Rasekh H, Rostami-Nejad M, Yousofi HA, Hosseini-Safa A. Molecular identification of Giardia lamblia; is there any correlation between diarrhea and genotyping in Iranian population? Gastroenterology and hepatology from bed to bench. 2014;7(3):168-72.

8. Adam RD. The Giardia lamblia genome. Int J Parasitol. 2000;30(4):475-84.

9. Johnston SP, Ballard MM, Beach MJ, Causer L, Wilkins PP. Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens. J Clin Microbiol. 2003;41(2):623-6.

10. Weiss JB, van Keulen H, Nash TE. Classification of subgroups of Giardia lamblia based upon ribosomal RNA gene sequence using the polymerase chain reaction. Mol Biochem Parasitol. 1992;54(1):73-86.

11. Skhal D, Aboualchamat G, Al Mariri A, Al Nahhas S. Prevalence of Giardia duodenalis assemblages and sub-assemblages in symptomatic patients from Damascus city and its suburbs. Infect Genet Evol. 2017;47:155-60.

12. Salman Y, Al-Taee A-R, Abid A. Prevalence of Giardia lamblia among Iraqi Displaced Peoples in Kirkuk Province ; Int.J.Curr.Microbiol.App.Sci (2016) 5(1): 753-760.

13. Arshad Ya. Prevalence of Intestinal Parasites (Entamoeba species and Giardia lamblia) in Duhok and Erbil cities, Northern Iraq. Journal of Microbiology & Experimentation. 2017;4(6).

14. Shalaby I, Gherbawy Y, Banaja A. Molecular characterization of Giardia parasite isolated from stool samples collected from different hospitals in Taif City (Saudi Arabia). 2011; Tropical biomedicine 28 (3); 487- 496.

15. Choy SH, Al-Mekhlafi HM, Mahdy MAK, Nasr NN, Sulaiman M, Lim YAL, et al. Prevalence and associated risk factors of Giardia infection among indigenous communities in rural Malaysia. Sci Rep. 2014;4:6909.

16. Faria CP, Zanini GM, Dias GS, da Silva S, Sousa MdC. Molecular Characterization of Giardia lamblia: First Report of Assemblage B in Human Isolates from Rio de Janeiro (Brazil). PLOS ONE. 2016;11(8).

17. Naguib D, El-Gohary AH, Roellig D, Mohamed AA, Arafat N, Wang Y, et al. Molecular characterization of Cryptosporidium spp. and Giardia duodenalis in children in Egypt. Parasit Vectors. 2018;11(1):403.

18. Júlio C, Vilares A, Oleastro M, Ferreira I, Gomes S, Monteiro L, et al. Prevalence and risk factors for Giardia duodenalis infection among children: A case study in Portugal. Parasites & Vectors. 2012;5(1):22.

19. Damitie M, Mekonnen Z, Getahun T, Santiago D, Leyns L. Molecular epidemiology of Giardia duodenalis infection in humans in Southern Ethiopia: a triosephosphate isomerase gene-targeted analysis. Infect Dis Poverty. 2018;7(1):17.

20. Kasaei R, Carmena D, Jelowdar A, Beiromvand M. Molecular genotyping of Giardia duodenalis in children from Behbahan, southwestern Iran. Parasitol Res. 2018;117(5):1425-31.

21. Jelinek T, Neifer S. Detection of Giardia lamblia stool samples: a comparison of two enzyme-linked immunosorbent assays.2013; F1000Research.2:39;1-6.

22. Al Saeed AT, Issa SH. Detection of Giardia lamblia antigen in stool specimens using enzyme-linked immunosorbent assay. 2010; EMHJ.Vol.16 No.4; 362-364.

23. Wilke H, Robertson LJ. Preservation of Giardia cysts in stool samples for subsequent PCR analysis. Journal of microbiological methods. 2009;78 (3):292-6.

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.