SOLID PHASE ASSAY FOR SALIVA

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This research project is designed to lead to the development of a system for the direct, rapid, reliable and accurate immunoassay of low molecular weight compounds in human saliva. The envisioned system is one that will be easy to use, will not require extraction or chromatography, and will give colorimetric endpoints that can be either qualitative and field- adaptable or, with suitable instrumentation, quantitative. The system will be built upon a novel device that serves to collect and process saliva automatically and unobtrusively yielding in a few minutes several mL's of clear, sterile, protein free filtrate containing compounds of low molecular weight. These compounds are often in equilibrium with the free fraction of the same substances in serum. The feasibility of this saliva collection device and a solid-phase assay system for progesterone were demonstrated during Phase I research. During Phase II it is proposed to extend these investigations to additional compounds, to combine the technologies, and to determine optimal conditions needed to demonstrate commercial feasibility of the approach. In particular during Phase II it is proposed to develop conditions for construction of the saliva collection system that will be acceptable to users and optimal for saliva collection, analyte processing, and colorimetric immunoassay performance; prepare derivatives and analyte- enzyme conjugates suitable for immunization and colorimetric immunoassay; develop monoclonal antibodies suitable for colorimetric immunoassay of the selected compounds; and develop and validate several colorimetric solid phase immunoassays for these compounds in human saliva. Immunoassay development during Phase II will focus on the steroids testosterone, progesterone, cortisol and estriol; the prostanoid thromboxane; and a major metabolite of cocaine, benzoylecgonine.