Lesion aspirate culture for the diagnosis and isolation of Leishmania spp. from patients with cutaneous leishmaniasis.
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.
Luz
Zélia Maria Profeta
ZM
Laboratório de Pesquisas Clínicas, Centro de Referência em Leishmanioses, Instituto René Rachou, Fiocruz, Belo Horizonte, MG, Brasil, 30190-002. profeta@cpqrr.fiocruz.br
Silva
Alexandre Rotondo da
AR
Silva
Fernanda de Oliveira
Fde O
Caligiorne
Rachel B
RB
Oliveira
Edward
E
Rabello
Ana
A
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
D016054:DNA, Protozoan