The diagnosis of cutaneous leishmaniasis (CL) may depend on the detection of the parasite in histologic sections, the growth of the promastigotes in culture, or the identification of parasite by other techniques. We performed polymerase chain reaction (PCR) on paraffin-embedded biopsies to determine the validity of this technique for diagnosis of CL. PCR was used to detect the parasite using 2 different DNA extraction methods. PCR was positive in all 20 cases when the Leishmania parasite was detected by light microscopy. Twenty-seven of 34 cases that were negative microscopically for the parasite were positive using PCR. The first extraction method of DNA identified leishmanial DNA in 41 of 54 cases (75.9%); the second extraction of DNA was positive in 47 of 54 cases (87%). PCR was negative in all of the nonleishmaniasis cases. The PCR-based method appears to be a useful diagnostic approach for identification of suspected cases of CL.