Tri-Elisa for Simultaneous Analysis of Three Analytes. Phase 1.

The goal of this research effort was to develop a three-enzyme ELISA system in which three analytes can be quantitated concurrently from a single biological or environmental sample. Three Plasmodium malaria antigens and anti-strain specific antibodies previously identified and optimized at the Walter Reed Army Institute of Research were used as a model system. During this Phase I effort, we identified three enzymes and a common substrate solution that will work simultaneously, providing three discernable colors as perceived by visual observation. We then conjugated each enzyme to a separate antibody, developed individual ELISAs for each analyte, and tested the three analyte systems concurrently. The resulting tri-ELISA system is a logical, efficient and potentially productive approach for improving an existing diagnostic assay for malarial sporozoites in mosquitos. The improved assay can be used by the World Health Organization for epidemiological studies designed to monitor, and through preventative mechanisms, potentially eradicate this disease which currently threatens approximately 400 million of the worlds population. The results of this Phase I project demonstrate feasibility and provide the basis for continuation into a Phase II project.