Rabbit monospecific antibody Leishmania major surface glycoprotein (gp63) was used in an indirect immunofluorescence assay (IFA) to identify promastigotes and amastigotes from sandflies and lesion fluid aspirates, respectively. Anti-gp63 fluoresced the entire periphery and flagella of promastigotes of different Leishmania species at dilutions to 1:50. Promastigotes were distinctively demonstrated in whole infected sandfly homogenate for 17 days post-infection. Amastigotes from lesion aspirates of infected BALB/c mice and from a human cutaneous leishmaniasis case were also fluoresced by the antibody. This technique could prove to be especially useful in arthropod vector surveillance efforts in which detection of the pathogen is as important as speciation of the sand fly vector.