SOLID PHASE ASSAY FOR SALIVA

这个研究项目旨在开发一个系统， 直接、快速、可靠、准确的低分子量 人类唾液中的化合物。 设想的系统将 易于使用，不需要提取或色谱法， 给出比色终点，可以是定性的，也可以是现场的， 适应性强，或者使用合适的仪器进行定量分析。 系统将 是建立在一个新的设备，用于收集和处理唾液， 在几分钟内自动且不引人注目地产生几毫升的 澄清、无菌、无蛋白质的滤液，含有低分子化合物 重量. 这些化合物通常与化合物的游离部分处于平衡。 血清中的相同物质。 这种唾液收集的可行性 装置和孕酮的固相分析系统被证明 在第一阶段的研究中。 在第二阶段，建议延长这些 研究其他化合物，联合收割机技术， 确定证明商业可行性所需的最佳条件 的方法。 特别是在第二阶段，建议开发 唾液收集系统的建设条件， 对于使用者来说是可接受的，并且对于唾液收集，分析物处理， 制备衍生物和分析物- 适用于免疫和比色免疫测定的酶缀合物; 开发适用于比色免疫测定的单克隆抗体 选择化合物;并开发和验证几种比色固体 这些化合物在人唾液中的阶段免疫测定。 免疫测定 在第二阶段的发展将集中在类固醇睾酮， 黄体酮、皮质醇和雌三醇;前列腺素类血栓素;以及主要的 可卡因的代谢物苯甲酰芽子碱。

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.