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Journal of Clinical Microbiology, October 2002, p. 3572-3576, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3572-3576.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Evaluation of PCR for Diagnosis of American Cutaneous Leishmaniasis in an Area of Endemicity in Northeastern Brazil

Eduardo Henrique Gomes Rodrigues,1 Maria Edileuza Felinto de Brito,1 Mitzi Guedes Mendonça,2 Roberto P. Werkhäuser,1 Eridan M. Coutinho,1 Wayner V. Souza,1 Maria de Fátima P. Militão de Albuquerque,1 Márcio L. Jardim,2 and Frederico G. C. Abath1*

Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz,1 Departmento de Dermatologia, Universidade Federal de Pernambuco, Recife, Brazil2

Received 11 February 2002/ Returned for modification 15 May 2002/ Accepted 30 June 2002

PCR-based approaches targeting kinetoplast DNA were evaluated for the diagnosis of American cutaneous leishmaniasis (ACL) in regions of endemicity in northeastern Brazil. A total of 119 cutaneous biopsy specimens from patients with ACL and nonleishmaniasis cutaneous lesions were studied. Two PCR-based systems were used; one was specific for the subgenus Viannia, and the other was specific for the genus Leishmania. The PCR specific for the subgenus Viannia had a sensitivity of 95.4%, whereas the genus-specific PCR detected the target DNA in 88.2% of the samples tested. The specificities of the assays, determined with samples from a group with nonleishmaniasis cutaneous lesions, was 100%. The results of the conventional tests indicate that the sensitivities of the PCR-based methods were significantly higher than those of smear examination, histological staining, and isolation by culture (P < 0.05). Antibodies specific for Leishmania braziliensis were detected by indirect immunofluorescence in 82.9% of the patients tested. Parasites were isolated from 40 of 86 patients (46.5%). Sixty-seven percent of dermal scrapings and 66.2% of stained tissue sections were positive by microscopy. Amplified products from the subgenus-specific PCR hybridized with the Leishmania panamensis minicircle, confirming infection consistent with L. braziliensis. The evidence available at present incriminates L. braziliensis as the only causative agent of ACL in the state of Pernambuco in Brazil.

* Corresponding author. Mailing address: Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, Av. Prof. Moraes Rego s/n, Cidade Universitária, 50670-420, Recife, Brazil. Phone: 55-81-33012558. Fax: 55-81-34532449. E-mail: fabath{at}cpqam.fiocruz.br.

Journal of Clinical Microbiology, October 2002, p. 3572-3576, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3572-3576.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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