Evaluation of a dot-immunoblot assay for detecting leishmanial antigen in naturally infected Phlebotomus argentipes (Diptera: Psychodidae).

A simple and highly reproducible dot-immunoblot assay was developed to detect leishmanial antigen in Phlebotomus argentipes that were naturally infected with Leishmania donovani. The test was sensitive to as little as 10 ng of antigenic protein (equivalent to the gut content of one laboratory-infected sandfly) and also appeared to be specific, in that it gave a positive result with some P. argentipes (the primary vector of L. donovani in India) and L. donovani but not with P. papatasi or other pathogens. When used to investigate a large number of sandflies collected from two areas of the Indian state of Bihar where visceral leishmaniasis is endemic, the assay appeared sufficiently sensitive and specific to detect the naturally infected insects. The simplicity, reproducibility, high sensitivity and high specificity of the assay should make it useful for field studies, particularly in determining the prevalence of sandfly infection, the local level of transmission, and the impact of vector-control programmes.