Volume 16, Issue 3 (May-Jun 2022)                   mljgoums 2022, 16(3): 24-29 | Back to browse issues page

XML Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Samimi A, Jorjani O N, Sharifi Z, Koohsar F, kalavi K, Mesgarian F et al . Detection of Leishmania major using PCR-ELISA. mljgoums. 2022; 16 (3) :24-29
URL: http://mlj.goums.ac.ir/article-1-1320-en.html
1- Department of Biotechnology, Faculty of New Technologies, Golestan University of Medical Sciences, Iran
2- Laboratory Science Research Center, Faculty of Paramedicine, Golestan University of Medical Sciences, Gorgan, Iran , niaz_jorjani@yahoo.com
3- Professor, Department of Virology, Iranian Blood Transfusion Organization, Tehran, Iran
4- Laboratory Sciences Research Center, Faculty of Paramedicine, Golestan University of Medical Sciences, Gorgan, Iran
5- Department of Medical Parasitology, Gonbad-e Kavus Health Center, Golestan University of Medical Sciences, Gorgan, Iran
6- Private veterinary physician, Gorgan,Iran
Abstract:   (348 Views)
Background and objectives: Cutaneous leishmaniasis is endemic in most areas of Iran, and the diagnosis of its species is essential for controlling the disease. Leishmania major is the causative agent for cutaneous leishmaniasis in humans. Molecular methods are generally more sensitive than microscopic methods. The present study aimed to use a polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) technique for detecting live L. major from wounds of patients with cutaneous leishmaniasis.
Methods: In the present study, a standard strain of L. major promastigotes was used as the positive control for purification of DNA. The Novy–MacNeal–Nicolle and RPMI-1640 media were used for reproduction of parasites. DNA was isolated from specimens taken from 35 patients with suspected cutaneous leishmaniasis whose disease was confirmed by direct smear method. The PCR-ELISA technique was later applied by using the standard strain, patient specimens, and primers specific for the 18s rRNA.
Results: Out of 35 patients, 17 (48.6%) were male and 18 (51.4%) were female. In addition, 8.6% of the patients lived in the Gonbad-e Kavus County, while all patients had been infected in villages around Gonbad-e Kavus. Of 35 patients with confirmed cutaneous leishmaniasis according to the direct smear method, 31 patients (86.31%) had leishmaniasis based on the PCR method and the PCR-ELISA methods.
Conclusion: Based on the results, the PCR-ELISA method is more sensitive and accurate for detecting L. major.
Full-Text [PDF 788 kb]   (118 Downloads) |   |   Full-Text (HTML)  (110 Views)  
Research Article: Original Paper | Subject: Parasitology
Received: 2020/08/31 | Accepted: 2020/09/5 | Published: 2022/05/14 | ePublished: 2022/05/14

References
1. Berman JD. Human leishmaniasis: clinical, diagnostic and chemotherapeutic developments in the last 10 years. Clin Infect Dis. 1997; 24(4): 684-703. [View at Publisher] [DOI:10.1093/clind/24.4.684] [PubMed] [Google Scholar]
2. Davies CR, Reithinger R, Campbell-Lendrum D, Feliciangeli D, Borges R, Rodriguez N. The epidemiology and control of leishmaniasis in Andean countries. Cad SaudePublica. 2000; 16(4): 925-50. [DOI:10.1590/S0102-311X2000000400013] [PubMed] [Google Scholar]
3. Rassi Y, Javadian E, Jalali M, Motazedian MH, Vatndoost H. Investigation on Zoonotic Cutaneous Leishm- aniasis,Southern Iran. Iranian Journal of Public Health. 2004; 33(1): 31-5. [Persian] [View at Publisher] [Google Scholar]
4. Saebi E. parasitic diseases in Iran. 4ed. Tehran: Aiish Publications. page 185. [Persian]
5. Mohebali M. Common Diseases of Humans and Animals. 1ed. Tehran: 6-62. [Persian]
6. Mesgarian F, Rahbarian N, Mahmoud Radi M, Hajaran H, Shahbaz F, Mesgarian Z, Taghipour N. Identification of Leishmania species isolated from human cutaneous Leishmaniasis in Gonbad-e-Qabus city using a PCR method during 2006-2007. Tehran Univ Med J. 2010; 68(4): 250-256.[Persian] [View at Publisher] [Google Scholar]
7. Amini Rad S, Amani J, Imani Fooladi AA, Saeedi P, MoosazadehMoghadam M. Detection of Acinetobacterbaumannii by PCR-ELISA method. J Shahrekord Univ Med Sci. 2017; 19(2): 6-16. [Persian]. [View at Publisher] [Google Scholar]
8. Sails AD, Bolton FJ, Fox AJ, WareingDRA,Greenway DLA. Detection of Campylobacter jejuni and Campylobacter coli in environmental waters byPCR enzyme-linked immunosorbent assay. Appl Environ Microbiol 2002; 68(3): 1319-24. [View at Publisher] [DOI:10.1128/AEM.68.3.1319-1324.2002] [Google Scholar]
9. Dehghan A, Ghahramani F, Hashemi B. Study on cutaneous leishmaniasis in Lorestan From 2005-2008. Journal ofJahromUiversity of Medical Sciences. 2009; 8(12): 7-11. [Persian]. [View at Publisher] [DOI:10.29252/jmj.8.3.8]
10. KarimianShirazi. Molecular identification of Leishmania species causingcutaneousleishmaniasis in Mashhad, Iran.Journal of Birjand University of Medical Sciences. 2014; 21 (2): 237-245.[Persian] [View at Publisher] [Google Scholar]
11. Silveira TG, Arraes SM, Bertolini DA, Teodoro U, Lonardoni MV, Roberto AC, et al. The laboratory diagnosis and epidemiology of cutaneous leishmaniasis in parana state, southern Brazil. Rev SocBras Med Trop. 1999; 32(4): 413-23. [DOI:10.1590/S0037-86821999000400013] [PubMed] [Google Scholar]
12. Jorjani O, lMirkarimi K, Charkazi A, Dadban Shahamatd Y, Mehrbakhshe Z, Bagheri A. The epidemiology of cutaneous leishmaniasis in Golestan Province, Iran: A cross-sectional study of 8-years. Parasite Epidemiol Control. 2019; 5: e00099. [View at Publisher] [DOI]
13. Tohidi F Barghae A. Cutaneous leishmaniasis Parasite Identification via PCR in the Infected Areas in Golestan Province. Knowledge & Health 2011; 6(2): 26-31.[Persian]. [View at Publisher] [Google Scholar]
14. Jorjani O, Kamalinia H, Mehrbakhsh Z, Ziaei-Hezarjaribi H, Pourrostami K, Mansourian M, Safari O, et al. Pediatric Cutaneous Leishmaniasis in Golestan Province, Iran: A Cross-Sectional Study of 8-years. International Journal of Pediatrics. 2019; 7(8): 9831-9839. [View at Publisher] [Google Scholar]
15. Gurel MS, Ulukanligil M, Ozbilge H. Cutaneous Leishmaniasis in sanliufa: epidemiologic and clincalfeatures of the last four years 1997-2000. Int J Dermatol 2002;41(1):32-7. [View at Publisher] [DOI:10.1046/j.0011-9059.2001.01396.x] [PubMed] [Google Scholar]
16. Saghafipour A, Akbari A, Rasi Y, Mostafavi R. Epidemiology of Cutaneous Leishmaniasis in Qom Province, Iran, during 2003-2009. Qom Univ Med Sci J. 2012; 6 (1) :83-88 [Persian]. [View at Publisher]
17. MR Jahani, MJ Gharavi, H Hadi Shirzad. Passive Detection of Cutaneous Leishmaniasis in Police Personnel Deployed in the Provinces of Isfahan, Ilam, Bushehr, Khorasan and Khuzestan, Iran. Iran J Public Health. 2003;32(3):23-27. [View at Publisher] [Google Scholar]
18. Mahmoodi MR, Tavakoli Afshar J , Mohajeri M , Fati AM ,Yazdanpanah MJ,Shakeri MT,Mirzaei A. Molecular Identification of Leishmania Species Causing Cutaneous Leishmaniasis in Mashhad, Iran. sjimu. 2010; 18 (2) :17-23. [Persian]. [View at Publisher]
19. Mohammad Hossein Feiz Haddad MH, Ghasemi E, Marghi Sh, Tavala M. Identification of Leishmania Species Isolated from Human Cutaneous Leishmaniasis in Mehran, Western Iran Using Nested PCR. Iran J Parasitol. 2016; 11(1): 65–72. [View at Publisher] [PubMed] [Google Scholar]
20. Al-Jawabreh A, Schoenian G, HamarshehO,Presber W. Clinical diagnosis of cutaneous leishmaniasis: A comparison study between standardized graded direct microscopy and ITS1-PCR of Giemsa-stained smears. Acta Trop. 2006; 99(1): 55-61. [View at Publisher] [DOI:10.1016/j.actatropica.2006.07.001] [PubMed] [Google Scholar]
21. Pagheh A. Detection and Identification of Causative Agent of CutaneousLeishmaniasis Using Specific PCR. J MazandUniv Med Sci. 2012; 22(Supple 1): 85-92. [Persian] [View at Publisher] [Google Scholar]
22. Fakhar M, Mikaeili F, Hatam GR, HabibiP,Karamian M, Motazedian MH, et al.Molecular epidemiology survey of cutaneous leishmaniasis in referral patients to Parasitology lab at Shiraz School of Medicine and importance application of PCR for diagnosis of disease. J JahromUniv Med Sci 2010;8(1): 1-5 [Persian]. [View at Publisher] [DOI:10.29252/jmj.8.1.2] [Google Scholar]
23. Perelle S, Dilasser F, Malorny B, Grout J, Hoorfar J, Fach P. Comparision of PCR-ELISA and LightCycler real-time PCR assays for detecting Salmonella spp. In milk and meat samples. Mol Cell Probes 2004; 18: 409-20 [View at Publisher] [DOI:10.1016/j.mcp.2004.07.001] [PubMed] [Google Scholar]
24. Ge B, Zhao S, Hall R, Meng J. A PCR-ELISA for detecting Shiga toxin-producing Escherichia coli. Microbes Infect. 2002; 4(3): 285-90. [View at Publisher] [DOI:10.1016/S1286-4579(02)01540-X] [PubMed] [Google Scholar]
25. Gomes LI, Dos Santos Marques LH, Enk MJ, de Oliveira MC, Coelho PM, Rabello A. Development and evaluation of a sensitive PCR-ELISA system for detection of schistosoma infection in feces. PLoS Negl Trop Dis. 2010; 4(4): e664. [View at Publisher] [DOI:10.1371/journal.pntd.0000664] [PubMed] [Google Scholar]
26. TetyanaKobets et al. Leishmania parasite detection and quantification using PCR-ELISA. Nature Protocols 5, 1074-1080 (1 June 2010) [View at Publisher] [DOI:10.1038/nprot.2010.68] [PubMed] [Google Scholar]

Add your comments about this article : Your username or Email:
CAPTCHA

Send email to the article author


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2007 All Rights Reserved | Medical Laboratory Journal

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