Disposable Nitrocellulose Filtration Plates Simplify the Dot-ELISA for Serodiagnosis of Visceral Leishmaniasis

摘要

This report describes a rapid, visually interpreted micro-enzyme immunoassy termed the Dot-ELISA for serodiangosis of visceral leishmaniasis (kala-azar) which uses 1.0 microliter volumes of parasite antigen adsorbed to nitrocellulose filter paper (antigen discs) and precipitable chromogenic substrate. This modified procedure was adapted form that of HAWKES et al. (1982), which was used to identify monoclonal antibodies. The Dot-ELISA is as sensitive and specific in detecting parasite-specific antibodies as conventional ELISA tests, is more rapid and requires much less antigen per test well. However, the procedure presently requires that antigen discs be cut and loaded into flat-bottom microtitre plates before testing. In order further to simplify the Dot-ELISA for laboratory and field use, we dotted disposable 96 well nitrocellulose filtration plates with 1.0 microliter formalin-fixed Leishmania donovani promastigote antigen per well and assayed a battery of sera from parasitologically proven patients and from healthy North Americans in the Dot-ELISA.