Development and optimization of the FAST-ELISA for detecting antibodies to Schistosoma mansoni

Abstract

The Falcon assay screening test (F.A.S.T. ®) system was used to develop a rapid, sensitive, and quantitative kinetic-based enzyme-linked immunosorbent assay (k-ELISA) for detecting antibodies against Schistosoma mansoni adult microsomal antigens (MAMAs). The FAST-ELISA uses polystyrene beads on sticks molded to the lid of a microtitration plate. The beads are coated with antigen. Reagents and sera are placed in microtitration plates and the beads exposed to reagents by immersion. The exposure time required for a single dilution of serum or other antibody source, conjugate, and substrate is 5 min each. Excluding preparation time, two plates can easily be assayed in 30 min. The optima for assay conditions, reproducibility, quantitative linearity, and sensitivity are delineated. A battery of sera from patients with both homologous and heterologous infections was tested, and a dilution series of a standard reference serum pool was included with each test. Results were expressed in number of units as calibrated against the standard reference sera pool. Antigen-coated bead storage studies were performed with untreated and three chemically treated antigens. The storage stability of MAMA, ability to perform the assay with minimal equipment, sensitivity, short assay time, and ease of operation make the FAST-ELISA ideal for field studies.