Articles

Identification of Leishmania Species Using PCR Assay on Giemsa-Stained Slides Prepared From Cutaneous Leishmaniasis Patients

Abstract

 

Background: Leishmaniasis is a group of diseases that are created by intracellular parasites of Leishmania. Cutaneous leishmaniasis is considered as one of the health problems in some provinces of Iran.

Methods: In this study, a total of 178 Giemsa-stained slides from confirmed cases of cutaneous leishmaniasis were examined. The slides were prepared from the patients with cutaneous leishmaniasis that referred to health centers and infected during the epidemic of cutaneous leishmaniasis in Poldokhtar city, Lorestan Province, Iran in 2006.Genomic DNA from each slide was extracted. After DNA extraction, ITS-PCR was used.

Results: Out of 178 slides, 129 (72.47%) samples had a band in the range of 485 bp and 49 (27.53%) samples 626 bp that matched L. tropica and L. major standard samples, respectively.

Conclusion: This study showed that Leishmania DNA could be efficiently ex-tracted and amplified even from old Giemsa-stained microscopic slides that were stored more than 6 yr. In this study was shown that both L. tropica and L. major species exist in Lorestan Province.

Bates PA. Transmission of Leishmania met acyclic promastigots by phlebotomine sand files. Int J Parasitol. 2007: 37(10-3)1097-1106.

WHO (2009) Leishmaniasis: the global trend. http:// www.who.into /neglected_ disease/ inte-grated-media_ Leishmaniasis/ en/ index. html.

World Health Organization (WHO) (2010). Con-trol of the leishmaniases. Report of a Meeting of the WHO Expert Committee on the Control of Leishmaniases, WHO Technical Report Series 949, Geneva. pp. 1-187.

Herwaldt BL, Magill AJ. Leishmaniasis, Cutane-ous, Infectious diseases related to travel, Centers for Disease Control and Prevention, Traveler’s health. Yellow book, 2012; Chapter.

Piscopo Tv, Mallia Azzopardi C. Leishmanias. Postqrad Med J. 2007; 83 (976):649-657.

Zoonoses Control Office, (2010). CDC, Ministry of Health and Medical Education, Iran.

Farahmand M, Nahrevanian H, Atashi Shirazi H, Naeimi S, Farzanehnejad Z. An overview of a di-agnostic and epidemiologic reappraisal of cuta-neous leishmaniasis in Iran. Braz J Infect Dis. 2011; 15 (1): 17-21.

Rahbarian N, Mesgarian A, Mahmoudi Rad M, Hajaran H, Shahbazi F, Mesgarian Z, Taghipour N. Identification of Leishmania species isolated from human cutaneous leishmaniasis using PCR method. J Res Health Sci. 2009; 9 (2): 48-51.

Singh S. New developments in diagnosis of Leishmaniasis. Indian J Med Res . 2006:123 (3): 311-30.

Tavares RG, Staggemeier R, Borges ALP, Rodri-gues, Castelan LA, Vasconcelos J, et al. Molecular techniques for the study and diagnosis of parasite infection. J Venom Anim Toxins incl Trop Dis. 2011; 17 (3): 239-248.

Banuls AL. Hide M, Prugnolle F. Leishmania and the Leishmaniases: A parasite genetic update and advances in taxonomy, epidemiology and patho-genicity in humans. Adv Parasitol. 2007; 64:1-109

Sharifi F, Sharifi I, Zarean M, Hakimi Parizi M, Aflatoonian MR, Fasihi Harandi M, Zahmatkesh R, Mashayekhi M, Kermanizadeh AR. Spatial dis-tribution and molecular identification of Leishmania species from endemic foci of south-eastern Iran. Iranian J Parasitol. 2012; 7: 45-52.

Yavar R, Abedin S, Reza AM, Ali OM, Sina R, Mehdi M, Reza YE, Fatemeh M, Babak F. Phlebo-tomus papatasi and Meriones libycus as the vector and reservoir host of cutaneous leishmaniasis in Qomrood district, Qom province, and central Iran. Asian Pac J Trop Med. 2011; 4: 97-100.

Shahbazi F, Shahabi S, Kazemi B, Mohebali M, Abadi AR, Zare Z. Evaluation of PCR assay in diagnosis and identification of cutaneous leishma-niasis: a comparison with the parasitological methods. Parasitol Res. 2008; 103: 1159-1162.

Vaeznia H, Dalimi A, Sadraei J, Pirstani M. Deter-mination of Leishmania species causing cutaneous leishmaniasis in Mashhad by PCR-RFLP method. Archives of Razi Institute. 2009; 64: 39-44.

Razmjou S, Hejazya H, Motazedianb MH, Baghaeia M, Emamyc M, Kalantaryb M. A new focus of zoonotic cutaneous leishmaniasis in Shi-raz, Iran. Trop Med Hyg. 2009; 103: 727-730.

Maraghi S, Samarbaf Zadeh A, Sarlak AA, Gha-semian, Vazirianzadeh B. Identification of cuta-neous leishmaniasis agents by Nested Polymerase Chain Reaction (Nested-PCR) in Shush city, Khuzestan province, Iran. Iranian J Parasitol. 2007; 2: 13-15.

Kheirandish F, Chegeni Sharafi A, Kazemi B, Bandehpour M, Tarahi MJ, Khamesipour A. First molecular identification of Leishmania species in a new endemic area of cutaneous leishmaniasis in Lorestan, Iran. Asian Pac J Trop Biomed. 2013; 6 (9): 713–717.

Chegeni Sharafi A, Amani H, Kayedi MH,Yarahahmadi A, Saki M, Mehrdad M, Nasiri E. Epidemiological Survey of Cutaneous Leishmaniasis in Lorestan Province(Iran) and In-troduction of Disease Transmission in New Lo-cal Areas. J IUMS. 2011; 19 (1): 54-60 (In Persian).

Gomes AH, Ferreira IM, Lima ML, Cunha EA, Garcia AS, Araujo MF, Pereira-Chioccola VL. PCR identification of Leishmania in diagnosis and control of canine leishmaniasis. Vet Parasitol. 2007; 144 (3-4): 234-241. 21. Alimoradi S, Hajjaran H, Mohebali M, Mansouri F. Molecular identification of Leishmania species isolated human cutaneous leishmaniasis by RAPD-PCR. Iranian J Publ Health. 2009; 38 (2): 44-50.

Schonian G Nasereddin A Dinse N. PCR diag-nosis and characterization of Leishmania in local and imported clinical samples. Diag Microbiol In-fect Dis 2003; 47(1): 349-58.

Eroglu F, Koltas IS, Genc A. Identification of Causative Species in Cutaneous Leishmaniasis Pa-tients Using PCR-RFLP. J Bacteriol Parasitol. 2011; 2:113. doi:10.4172/2155-9597.1000113.

Marfurt J, Nasereddin A, Niederwieser I, Jaffe CL, Beck HP, Felger I . Identification and diffe-rentiation of Leishmania species in clinical samples by PCR amplification of the Miniexon sequence and subsequent Restriction Fragment Length Polymorphism analysis. J Clin Microbiol. 2003; 41(7): 3147-53.

Dweik A, Schonian G, Mosleh IM, Karanis P. Evaluation of PCR_RFLP (based on ITS-1 and HeaIII) for the detection of Leishmania species, using Greek canine isolates and Jordanian clinical material. Ann Trop Med Parasitol 2007:-101(5):399-407.

Martínez LP, Rebollo JA, Luna AL, Cochero S, Bejarano EE. Molecular identification of the par-asites causing cutaneous leishmaniasis on the Car-ibbean coast of Colombia. Parasitol Res. 2010; 106 (3): 647-652.

Motazedian H, Karamian M, Noyes HA, Arde-hali S. DNA extraction and amplification of Leish-mania from archived, Giemsa-stained slides, for the diagnosis of cutaneous leishmaniasis by PCR. Ann Trop Med Parasitol. 2002; 96(1):31-34.

Kazemi-Rad E, Mohebali M, Hajaran H, Rezeai S, Memishi S. Diagnosis and characterization of Leishmania species in Giemsa-stained slides by PCR-RFLP. Iranian J Publ Health. 2008:37(1): 454-60.

Mahdy MAK, Al-Mekhlafi HM, Al-Mekhlafi AM, Lim YAL, Bin Shuaib NOM, Azazy AA, Mahmud R. Molecular Characterization of Leish-mania Species Isolated from Cutaneous Leishmaniasis in Yemen. PLoS ONE. 2010; 5: e12879. doi:10.1371/journal.pone.0012879.

Volpini AC, Marques MJ, Lopes dos Santos S, Machado-Coelho GL, Mayrink W, Romanha AJ. Leishmania identification by PCR of Giemsa-stained lesion imprint slides stored for up to 36 years. Clin Microbiol Infec. 2006; 12: 815-818.

Khademvatan S, Neisi N, Maraghi S, Saki J. Di-agnosis and identification of Leishmania spp. from Giemsa-stained slides, by real-time PCR and melting curve analysis in south-west of Iran. Ann Trop Med Parasitol. 2011;105:559-65.

Yvone Maia Brustoloni, Rosimar Batista Lima, Rivaldo Venâncio da Cunha, Maria Elizabeth Dorval, Elisa Teruya Oshiro, Ana Lúcia Lyrio de Oliveira, Claude Pirmez. Sensitivity and specificity of polymerase chain reaction in Giemsa-stained slides for diagnosis of visceral leishmaniasis in children. Mem Inst Oswaldo Cruz, Rio de Janeiro. 2007; 102: 497-500.

A. Al-Jawabreh, G. Schoenian, O. Hamarsheh, W. Presber. Clinical diagnosis of cutaneous leish-maniasis: A comparison study between stan-dardized graded direct microscopy and ITS1-PCR of Giemsa-stained smears. Acta Trop. 2006; 99: 55–61.

Alavinia SM, Arzamani K, Reihani MH, Jafari J. Some Epidemiological Aspects of Cutaneous Leishmaniasis in Northern Khorasan Province, Iran. Iranian J Arthropod-Borne Dis. 2009; 3(2).

Rassi Y, Javadian E, Amin M, Rafizedeh S, vatandoost H, Motazedian H. Meriones libycus is the main reservoir of zoonotic cutaneous leishmaniasis in south of Islamic Republic of Iran. East Mediterr Health J. 2006; 12 (3-4): 474-7.

Rassi Y, Sofizadeh A, Abai MR, Oshaghi MA, Rafizadeh S, Mohebail M, et al. Molecular detec-tion of L. major in the vectores and reservoir hosts of cutaneous leishmaniasis in Kalaleh district, Golestan Province, Iran. Iranian J Arthropod-Borne Dis. 2008; 2 (2): 21-27.

Hajjaran H, Mohebali M, Alimoradi S, Abai MR, Edrissian GhH. Isolation and characterization of pathogenic Leishmania turanica from Nesokia indica (Rodentia, Muridea) by PCR-RFLP and ITSI se-quencing in Iran. Trans R Soc Trop Med Hyg. 2009; 103 (11): 1177-9.

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IssueVol 8 No 3 (2013) QRcode
SectionArticles
Keywords
Cutaneous leishmaniasis Giemsa Iran Leishmania PCR

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How to Cite
1.
Kheirandish F, Chegeni Sharafi A, Kazemi B, Mohebali M, Sarlak A, Tarahi MJ, Holakouee K, Hajaran H. Identification of Leishmania Species Using PCR Assay on Giemsa-Stained Slides Prepared From Cutaneous Leishmaniasis Patients. Iran J Parasitol. 1;8(3):382-388.