Acta Tropica 79 (2001) 225–229 Sensitivity of the polymerase chain reaction for the diagnosis of cutaneous leishmaniasis due to Leishmania (Viannia ) guyanensis Gustavo A.S. Romero a, *, Marcus V.F. Guerra b , Marcilene G. Paes b , Elisa Cupolillo c , Cristiane Bentin Toaldo c , Vanize O. Mace ˆdo a , Octavio Fernandes d a Nu ´cleo de Medicina Tropical, Uniersidade de Brası ´lia, Campus Uniersita ´rio, Asa Norte, P.O. Box 04517, Brası ´lia, DF. 70919 -970, Brazil b Fundac ¸a ˜o de Medicina Tropical, Instituto de Medicina Tropical do Amazonas, Aenida Pedro Teixeira S /N, Manaus, Amazonas 69040 -000, Brazil c Departamento de Imunologia, FIOCRUZ, Aenida Brasil 4365, Manguinhos, Rio de Janeiro, RJ 21045 -900, Brazil d Departamento de Medicina Tropical, Instituto Oswaldo Cruz, FIOCRUZ, Aenida Brasil 4365, Manguinhos, Rio de Janeiro, RJ 21045 -900, Brazil Received 15 July 2000; received in revised form 2 February 2001; accepted 16 February 2001 Abstract The sensitivity of the polymerase chain reaction (PCR) in 35 consecutive outpatients with cutaneous leishmaniasis caused by Leishmania (Viannia ) guyanensis was evaluated using, as gold standard, the in vitro isolation of the parasite through culture of aspirates of the cutaneous ulcers. All isolates were identified using electrophoretic enzyme analysis. Patients were mainly young males with recent onset disease without prior specific treatment. PCR was performed using DNA extracted from fresh frozen biopsies of cutaneous ulcers. The reaction used a pair of oligonucleotides that amplify the conserved region of the minicircle molecule. PCR showed 100% sensitivity (95% CI from 90.0 to 100.0). These results were similar to the visualization of amastigotes in imprint preparations of cutaneous biopsy tissue and the inoculation of biopsy material in golden hamsters. Despite the high sensitivity of the PCR, in this particular clinical setting of cutaneous leishmaniasis caused by L.(V.) guyanensis in the Brazilian Amazon, it appears that the method of choice for diagnosis should be the direct visualization of amastigotes using imprint preparations and the PCR reserved for those patients with negative imprint results. © 2001 Elsevier Science B.V. All rights reserved. Keywords: Cutaneous leishmaniasis; Leishmania (Viannia ) guyanensis ; Diagnosis; Polymerase chain reaction; Brazil www.parasitology-online.com * Corresponding author. Tel.: +55-61-2735003; fax: +55-61-2732811. E-mail addresses: gromero@unb.br (G.A.S. Romero), imtam@prodamnet.com.br (M.V.F. Guerra), imtam@prodamnet.com.br (M.G. Paes), ecupoli@ioc.fiocruz.br (E. Cupolillo), ctoaldo@dbbm.gene.fiocruz.br (C. Bentin Toaldo), tropical@unb.br (V.O. Mace ˆdo), octaviof@dbbm.gene.fiocruz.br (O. Fernandes). 0001-706X/01/$ - see front matter © 2001 Elsevier Science B.V. All rights reserved. PII:S0001-706X(01)00140-1