Direct enzyme-linked immunosorbent assay: a simple immunoassay using Leishmania donovani promastigote for diagnosis of kala-azar.

For immunodiagnosis of kala-azar enzyme-linked immunosorbent assay (ELISA) and immunofluorescence testing (IFAT) are commonly used. In IFAT, whole parasite antigen and in ELISA the soluble antigen have been used. Preparation of ELISA antigen has certain inherent difficulties. We have developed a simple, specific, and quantitative immunoassay, "direct ELISA" for diagnosis of kala-azar. Intact formalinized promastigote suspension has been used to combine with the antibodies of the patient sera. The colour developed in the supernatant by the enzyme conjugate combined on the parasite surface was measured with a spectrophotometer. The test was able to detect kala-azar-specific antibodies at very high serum dilution and could discriminate between kala-azar and the common diseases prevalent in Asia. The optical densities of the sera of different control groups were significantly low. The method has potential for use as a diagnostic tool in less well equipped laboratories.