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Diagnosis of human visceral leishmaniasis
by PCR using blood samples spotted on
filter paper
Eduardo Sergio da Silva1, Célia Maria Ferreira Gontijo2, Raquel da Silva Pacheco3 and Reginaldo Peçanha Brazil3
1Universidade do Estado de Minas Gerais, Fundação Educacional de Divinópolis, Av. Paraná s/n,
Campus Universitário, 35500-970 Divinópolis, MG, Brasil
2Laboratório de Leishmanioses, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz,
Belo Horizonte, MG, Brasil
3Departamento de Bioquímica e Biologia Molecular, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
Corresponding author: E.S. Silva
E-mail: biologia@funedi.edu.br
Genet. Mol. Res. 3 (2): 251-257 (2004)
Received October 18, 2003
Accepted March 26, 2004
Published May 25, 2004

ABSTRACT. The polymerase chain reaction (PCR) is a simple, rapid procedure that has been adapted for the diagnosis of leishmaniasis. In the present study, 85 blood samples and seven bone marrow aspirates from 85 patients with clinical symptoms suggestive of visceral leishmaniasis from the metropolitan region of Belo Horizonte in the Brazilian State of Minas Gerais were screened using molecular and serological techniques. Samples that were negative (N = 12) and positive (N = 19) in parasitological and serological tests were used as controls. Of the 85 samples analyzed by PCR, 61 (71.7%) showed the expected amplification products in agarose gels. However, when the technique was combined with molecular hybridization, 72 samples (83.5%) gave a positive signal on film. Nineteen patients with Leishmania parasites in bone marrow cultures (positive controls) showed PCR hybridization in whole-blood samples, as did the seven bone marrow aspirates positive for Leishmania. None of the negative controls reacted in PCR or in an indirect immunofluorescent assay. These results indicate that PCR could replace the conventional parasitological examination in the diagnosis of leishmaniasis since it provides very satisfactory results with blood samples spotted on filter paper.

Key words: Diagnosis, Filter paper, Leishmania chagasi, Serological analysis, Visceral leishmaniasis

 

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