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Journal of Clinical Microbiology, June 1999, p. 1819-1823, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Use of PCR in Diagnosis of Human American Tegumentary Leishmaniasis in Rio de Janeiro, Brazil

Claude Pirmez,1,* Valéria da Silva Trajano,1 Manoel Paes-Oliveira Neto,2 Alda Maria da-Cruz,3 Sylvio Celso Gonçalves-da-Costa,3 Marcos Catanho,1 Wim Degrave,1 and Octavio Fernandes5

Department of Biochemistry & Molecular Biology,1 Evandro Chagas Hospital,2 Department of Protozoology,3 and Department of Tropical Medicine,5 Oswaldo Cruz Institute, and Department of Pathology, University of the State of Rio de Janeiro,5 Rio de Janeiro, Brasil

Received 3 November 1998/Returned for modification 17 December 1998/Accepted 12 March 1999

In Brazil, the most common etiological agent of American tegumentary leishmaniasis is Leishmania (Viannia) braziliensis. In general, diagnostic techniques envisage the visualization of the parasite, but that technique has a low sensitivity. The main purpose of the present work was to evaluate the PCR as a routine tool for the diagnosis of leishmaniasis. Biopsy specimens from cutaneous or mucosal lesions were taken from 230 individuals from areas where Leishmania is endemic: 216 patients who had a clinical picture suggestive ofl leishmaniasis and 14 individuals with cutaneous lesions due to other causes. Each specimen was processed for histopathologic examination, culture, touch preparation, and DNA isolation. Oligonucleotides that amplify the conserved region of the minicircle molecules of Leishmania were used in a hot-start PCR. While at least one conventional technique was positive for Leishmania for 62% (134 of 216) of the patients, PCR coupled to hybridization was positive for 94% (203 of 216) of the patients. The 14 patients whose clinical picture was not suggestive of leishmaniasis had negative results by all techniques. The impact of the PCR was striking in mucosal disease. While the disease in only 17% (4 of 24) of the patients could be diagnosed by conventional techniques, PCR was positive for 71% (17 of 24) of the patients. Hybridization showed that all cases of disease were caused by parasites belonging to the Viannia subgenus. Altogether, the results indicate that PCR is a valuable tool for the diagnosis of leishmaniasis on a routine basis and is likely to provide valuable epidemiological information about the disease in countries where it is endemic.

* Corresponding author. Mailing address: Department of Biochemistry & Molecular Biology, Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil 4365, CEP 21045-900, Rio de Janeiro, RJ, Brazil. Phone: (55 21) 598-4349. Fax: (55 21) 590-3495. E-mail: pirmez{at}gene.dbbm.fiocruz.br.

Journal of Clinical Microbiology, June 1999, p. 1819-1823, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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