COLORIMETRIC MEASUREMENT OF MILK PROGESTERONE

这个第二阶段的SBIR应用提出了建立在一种新的、 固相、比色免疫分析系统用于定量 奶牛乳中孕酮的测定，方法的可行性为 在第一阶段研究中显示。浓度之间有很好的相关性 用常规方法测定牛奶中孕酮的含量 放射性标记示踪剂和新开发的酶免疫分析已经被 获得。该检测系统是基于使用能够 通过一种专有方法共价连接到微板孔 与非放射性、基于酶的信号发生器相结合。阶段 Ii研究的目的是将这种分析方法开发成一种简单、可靠的方法 发情周期、产后卵巢周期监测系统， 卵巢功能障碍，以及可以进行的奶牛怀孕 在野外条件下由未受过训练的人员执行。设想的系统是 一种易于使用、不需要提取或 层析，并将给出快速的定量结果。量化 将通过手动色度计完成，该色度计可以耦合 直接连接到运行定制软件的计算机。该系统将建成 基于一种用于收集和处理的新型设备(患者待定) 在几分钟内自动产生几毫升的液体 透明、无蛋白质的滤液，含有低分子化合物 重量。在第二阶段，旨在简化 将开发测试运行的方式，将进行测试 用于现场的健壮，对如此修改的进一步验证 将寻求化验，它在牛棚中的用途将是 演示的、能够吸引第三阶段支持的原型系统 将发展商业化。

This Phase II SBIR application proposes research that builds on a novel, solid phase, colorimetric immunoassay system for the quantitative measurement of progesterone in milk of cows, the feasibility of which was shown during Phase I research. A good correlation between concentrations of progesterone in milk measure by a conventional method using radiolabeled tracer and the newly developed enzyme immunoassay has been obtained. The assay system is based on use of monoclonal antibodies that are covalently linked by a proprietary method to microplate wells in combination with a non-radioactive, enzyme-based signal generator. Phase II research will be aimed at developing the assay into a simple, reliable system for monitoring the estrous cycle, ovarian cyclicity post partum, ovarian dysfunction, and pregnancy in dairy cows which can be performed under field conditions by a non-trained person. The envisioned system is one that will be easy to use, will not require extraction or chromatography, and will give rapid, quantitative results. Quantitation will be accomplished with a manual colorimeter that can be coupled directly to a computer running custom software. The system will be built upon a novel device (patient pending) that serves to collect and process fluids automatically yielding in a few minutes several milliliters of clear, protein-free filtrate containing compounds of low molecular weight. During Phase II, an integrated approach designed to simplify the manner that the assay is run will be developed, the assay will be made robust for use in the field, further validation for the thus modified assay will be sought, its usefulness in a dairy barn will be demonstrated, and prototype systems able to attract phase III support for commercialization will be developed.