title: Lesion aspirate culture for the diagnosis and isolation of Leishmania spp. from patients with cutaneous leishmaniasis.
abstract: The detection of Leishmania spp. in skin lesion aspirates, using a puncture technique, was evaluated in 76 patients with cutaneous leishmaniasis (CL) who were referred to a Leishmaniasis Reference Centre in Brazil. CL was defined based on skin lesions suggestive of the disease and on a positive result of the Montenegro skin test or Giemsa-stained imprints of biopsy fragments. The aspirates were cultured using a vacuum tube device containing culture medium and evaluated for the presence of Leishmania spp. The biphasic medium culture was examined once a week for three weeks. Promastigotes were observed in 53/76 (69.7%) cultures. Stained smears from 60 of the 76 patients were evaluated using PCR-RFLP to detect the conserved minicircle region of Leishmania spp. and to classify the parasite. Of these patients, 45 (75%) showed positive results in aspirate culture and 15 presented negative results. The PCR was positive in 80% (53/60) samples. The PCR-RFLP profile was determined in 49 samples, of which 45 (92%) showed a pattern compatible with Leishmania (Viannia) braziliensis. The aspirate culture is a sensitive and feasible method for diagnosing CL and may be routinely adopted by health services for L. (V.) braziliensis isolation and identification.
authors.affiliation: Laboratório de Pesquisas Clínicas, Centro de Referência em Leishmanioses, Instituto René Rachou, Fiocruz, Belo Horizonte, MG, Brasil, 30190-002. profeta@cpqrr.fiocruz.br
authors.forename: Zélia Maria Profeta
authors.initials: ZM
authors.lastname: Luz
authors.forename: Alexandre Rotondo da
authors.initials: AR
authors.lastname: Silva
authors.forename: Fernanda de Oliveira
authors.initials: Fde O
authors.lastname: Silva
authors.forename: Rachel B
authors.initials: RB
authors.lastname: Caligiorne
authors.forename: Edward
authors.initials: E
authors.lastname: Oliveira
authors.forename: Ana
authors.initials: A
authors.lastname: Rabello
mesh_terms: D000328:Adult; D000818:Animals; D046508:Culture Techniques; D016054:DNA, Protozoan / Q000032:analysis; D005260:Female; D006801:Humans; D007892:Leishmania braziliensis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D007894:Leishmania mexicana / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000469:parasitology; D008297:Male; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012882:Skin Tests
chemical_list: D016054:DNA, Protozoan