title: Optimization of microculture and evaluation of miniculture for the isolation of Leishmania parasites from cutaneous lesions in Peru.
abstract: Traditional culture of Leishmania parasites is labor-intensive and shows poor sensitivity. We evaluated microculture and novel miniculture methods for diagnosis of cutaneous leishmaniasis (CL). Consecutive patients who came to the Leishmaniasis Clinic, Hospital Nacional Cayetano Heredia, Lima, Peru, were enrolled. Lesion aspirates were cultured in traditional tubes containing Novy-MacNeal-Nicolle medium and in miniculture tubes (Eppendorf, Hamburg, Germany) and capillary tubes (microculture) containing RPMI 1640 medium containing 20% fetal bovine serum. The reference standard was positive results in two of four tests (smear, culture, polymerase chain reaction, or leishmanin skin test). Outcome measures were sensitivity and time to positivity. Fifty-five patients with 74 lesions were enrolled. Of 59 lesions that fulfilled reference criteria for CL, 50 were positive by microculture (sensitivity=84.7%; P=0.001), 45 by miniculture (sensitivity=76.3%; P=0.042), and 35 by traditional culture (sensitivity=59.3%). Median time to positivity was three days by microculture and miniculture and five days by traditional culture (P<0.001). Microculture and miniculture are sensitive and efficient means of diagnosing CL.
authors.affiliation: Department of Laboratory Medicine and Pathobiology, University of Toronto, and Tropical Diseases Unit, Toronto General Hospital, Toronto, Ontario, Canada. andrea.boggild@utoronto.ca
authors.forename: Andrea K
authors.initials: AK
authors.lastname: Boggild
authors.forename: Cesar
authors.initials: C
authors.lastname: Miranda-Verastegui
authors.forename: Diego
authors.initials: D
authors.lastname: Espinosa
authors.forename: Jorge
authors.initials: J
authors.lastname: Arevalo
authors.forename: Dalila
authors.initials: D
authors.lastname: Martinez-Medina
authors.forename: Alejandro
authors.initials: A
authors.lastname: Llanos-Cuentas
authors.forename: Donald E
authors.initials: DE
authors.lastname: Low
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002648:Child; D002675:Child, Preschool; D046508:Culture Techniques / Q000379:methods*; D005260:Female; D006801:Humans; D007891:Leishmania / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology; D008297:Male; D008875:Middle Aged; D010568:Peru / Q000453:epidemiology; D012680:Sensitivity and Specificity; D012867:Skin / Q000469:parasitology*