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Journal of Clinical Microbiology, November 2007, p. 3680-3684, Vol. 45, No. 11
0095-1137/07/$08.00+0     doi:10.1128/JCM.01286-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru{triangledown}

Andrea K. Boggild,1* Cesar Miranda-Verastegui,2 Diego Espinosa,2 Jorge Arevalo,2,3 Vanessa Adaui,2 Gianfranco Tulliano,2 Alejandro Llanos-Cuentas,2,4 and Donald E. Low1,5

Dept. of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada,1 Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru,2 Departamento de Bioquimica, Biologia Molecular y Farmacologia, Facultad de Ciencias, Universidad Peruana Cayetano Heredia, Lima, Peru,3 Organismo Andino de Salud, Convenio Hipolito Unanue, Lima, Peru,4 Laboratories Branch, Ontario Ministry of Health and Long-term Care, Etobicoke, Canada5

Received 26 June 2007/ Returned for modification 22 August 2007/ Accepted 10 September 2007

Traditional culture of Leishmania spp. is labor intensive and has poor sensitivity. We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for evaluation of skin lesions. Lesion aspirates were cultured in duplicate and parallel in traditional culture tubes containing modified Novy-MacNeal-Nicolle (NNN) medium or Roswell Park Memorial Institute medium 1640 with 10% fetal bovine serum (10% RPMI) and in 70-µl capillary tubes containing a mixture of lesion aspirate and 10% RPMI. For sensitivity analysis, the consensus standard was considered to be a positive result in any two of the following four tests: Giemsa-stained lesion smear, culture, kinetoplast DNA PCR, or leishmanin skin test. The outcome measures were sensitivity and time to culture positivity. Forty-five patients with 62 skin lesions were enrolled in the study, of which 53 lesions fulfilled the consensus criteria for a final diagnosis of cutaneous leishmaniasis. Of these 53 lesions, 39 were culture positive: 38 in capillary tubes, 29 in traditional culture tubes with modified NNN medium, and 19 in traditional culture tubes with 10% RPMI medium. The sensitivity of microculture was 71.7%, versus 54.7% for traditional culture with NNN (P, 0.038) and 35.8% with 10% RPMI (P, <0.001). The mean times to culture positivity were 4.2 days by microculture, 5.2 days in NNN, and 6 days in 10% RPMI (P, 0.009). We have demonstrated that microculture is a more sensitive and time-efficient means of isolating Leishmania parasites from cutaneous lesions than traditional culture.

* Corresponding author. Mailing address: Department of Microbiology, Mount Sinai Hospital, 600 University Avenue, Room 1485, Toronto, ON M5G 1X5. Phone: (416) 586-4695. Fax: (647) 439-0827. E-mail: andrea.boggild{at}utoronto.ca

{triangledown} Published ahead of print on 19 September 2007.

Journal of Clinical Microbiology, November 2007, p. 3680-3684, Vol. 45, No. 11
0095-1137/07/$08.00+0     doi:10.1128/JCM.01286-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Espinosa, D., Boggild, A. K., Deborggraeve, S., Laurent, T., Valencia, C., Pacheco, R., Miranda-Verastegui, C., Llanos-Cuentas, A., Leclipteux, T., Dujardin, J.-C., Buscher, P., Arevalo, J. (2009). Leishmania OligoC-TesT as a Simple, Rapid, and Standardized Tool for Molecular Diagnosis of Cutaneous Leishmaniasis in Peru. J. Clin. Microbiol. 47: 2560-2563 [Abstract] [Full Text]  
  • Boggild, A. K., Miranda-Verastegui, C., Espinosa, D., Arevalo, J., Martinez-Medina, D., Llanos-Cuentas, A., Low, D. E. (2008). Optimization of Microculture and Evaluation of Miniculture for the Isolation of Leishmania Parasites from Cutaneous Lesions in Peru. Am J Trop Med Hyg 79: 847-852 [Abstract] [Full Text]  


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