Molecular Identification of Leishmania Species Using Samples Obtained from Negative Stained Smears
-
Ma Mohaghegh
,
A Fata
,
Gh Salehi
,
F Berenji
,
M Mousavi Bazzaz
,
H Rafatpanah
,
M Parian
,
A Movahedi
Abstract
Background: Cutaneous Leishmaniasis (CL) is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers sus-pected to CL by PCR method.
Methods:Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, negative Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed.
Results:Among the DNA extracted from smears, L. tropica was isolated from 9 (11.1%) of the smears and L.major was not isolated from any samples.
Conclusion:Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accu-rate. PCR is recommended for clinically suspected lesions with negative result of direct smear.
Adhya S, Chatterjee M, Hassan MQ, Mukher-jee S, Sen S. Detection of Leishmania in the blood of early kala-azar patients with the aid of polymerase chain reaction. Trans R Soc Trop Med Hyg. 1995; 89: 622–624.
Blum J, Desjeux P, Schwartz E, Beck B, Hatz C. Treatment of cutaneous leishmaniasis among travellers. J Antimicrob Chemother. 2004; 53:158–166.
Magill AJ. Cutaneous leishmaniasis in the re-turning traveler. Infect Dis Clin N Am. 2005; 19:241-66.
Vega-Lopez F. Diagnosis of cutaneous leishmaniasis. Curr Opin Infect Dis. 2003; 16:97.
Rioux JA, Lanotte G, Serres E, Pratlong F, Bastien P, Perieres J. Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification. Ann Parasitol Hum Comp. 1990; 65:111-125.
Aviles H, Belli A, Armijos R, Monroy F, Har-ris E. PCR detection and identification of Leishmania parasites in clinical specimens in Ecuador: a comparison with classical diagnos-tic methods. J Parasitol. 1999; 85:181–187.
Bhattacharyya R., Das K, Sen S, Roy S, HK. Majumder. Development of a genus specific primer set for detection of Leishmania para-sites by polymerase chain reaction. FEMS Microbiol Lett. 1996; 135:195–200.
Martinez L, Rebollo JA, Luis Luna A, Co-chero S, Bejarano EE. Molecular identifica-tion of the parasites causing cutaneous leishmaniasis on the Caribbean coast of Co-lombia. Parasitol Res. 2010; 106(3):647-655.
Kazemi-Rad E, Mohebali M, Hajjaran H, Re-zaei S, Mamishi M. Diagnosis and Characteri-zation of Leishmania Species in Giemsa-Stained slides by PCR-RFLP. Iranian J Publ Health. 2008; 37(1):54-60.
Motazedian MH, karamian M, Ardahali S, Hanjani F. Evaluation of polymerase chain re-action method in identifying the species of Leishmania parasites in the Archive stained slides with Giemsa. Med Res J. 2004; 2(4):1-7. 11. Elahi R, Fata A, Berenji F. Comparison of la-boratory techniques in diagnosis of cutaneous leishmaniasis. Med J Mashhad Univ Med Sc. 1995; 47(1): 62-68.
Fata A, Khamesipour A, Mohajery M, Hos-seini- nejad Z, Afzal-aghaee M, Berenji F, Akhavan A, Ganjbakhsh M. Amin-Moham-madi A.Whatman paper (FTA cards) for Stor-ing and Transferring Leishmania DNA for PCR Examination. Iranian J Parasitol. 2009; 4(4):37-42.
Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL. Comparison of PCR assays for diagnosis of cutaneous leishmaniasis. J Clin Microbiol. 2006; 44:1435-9.
Rodrigues E, De Brito M, Mendonca M. Werkhauser RP, Coutinho EM, Souza WV, et al. Evaluation of PCR for diagnosis of Ameri-can cutaneous leishmaniasis in an area of endemicity in Northeastern Brazil. J Clin Microbiol. 2002; 40:3572-6.
Rodriguez N, Guzman B, Rodas A, Takiff H, Bloom AR, Convit J. Diagnosis of cutaneous leishmaniasis and species discrimination of parasites by PCR and hybridization. J Clin Microbiol. 1994; 32:2246-52. 16. Motazedian MH, Karamian M, Ardahali S, Hanjani F. Evaluation of polymerase chain re-action method in identifying the species of Leishmania parasites in the Archive stained slides with Giemsa. Med Res J. 2004; 2(4):1-7. 17. Mahboudi F, Abolhasani M, Tehrani SR, Azimi M. Diffrentation of old and new world Leishmania species at complex and species le-vels by PCR. Scand J Infect Dis. 2002;:34(10):756-758. 18. Omidian M, Khosravi AD, Nazari M, Rashidi A. The comparison of histopathological find-ings and polymerase chain reaction in lesions with primary clinical diagnosis of cutaneous leishmaniasis with negative smear. Pak J Med Sci. 2008; 24(1):96-99.
Safaei A, Motazedian MH, Vasei M. Polyme-rase chain reaction for diagnosis of cutaneous leishmaniasis in histologically positive, suspi-cious and negative skin biopsies. Dermatol. 2002; 205(1):18-24. 20. Fata A, Dalimi-asl A, Jafari MR, Mohajery M, Khamesipour A, Valizadeh M. Clinical appear-ance, Leishmanin Skin Test and ELISA using Monoclonal Antibody in diagnosis of different forms of Cutaneous Leishmaniasis. Med J Mashhad Univ Med Sc. 2004; 47(1): 19-27. 21. Mahmoodi MR, Mohajery M, Tavakkol Af-shari J, Taghae Shakeri M, Yazdan Panah MJ, Berenji F, Fata A. Molecular identification of Leishmania species causing cutaneous leishmaniasis in Mashhad, Iran. Jundishapur Journal of Microbiology. 2011; 3(4):195-200.
Valizadeh M, Dalimi Asl AH, Fata A, et al, A Study on Leishmania species Causing Cutane-ous Leishmaniasis in Mashhad Using Specific Monoclonal Antibodies, Modarres J Med.Sc. 2005, 7(2):107-113 .
| Files | ||
| Issue | Vol 8 No 2 (2013) | |
| Section | Articles | |
| Keywords | ||
| Cutaneous Leishmaniasis Direct smear Leishmania tropica PCR | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
