COLORIMETRIC MEASUREMENT OF MILK PROGESTERONE

这个第二阶段SBIR应用程序提出了一个新的， 固相，比色免疫测定系统的定量 测定奶牛乳汁中孕酮的可行性， 在第一阶段的研究中。浓度之间的良好相关性 用常规方法测量牛奶中孕酮的含量 放射性标记示踪剂和新开发的酶免疫测定法已被 得到了该测定系统基于使用单克隆抗体， 通过专有方法共价连接到微孔板威尔斯孔中， 与非放射性的、基于酶的信号发生器组合。相 II研究的目的将是发展成为一个简单，可靠的检测 用于监测发情周期、产后卵巢周期 卵巢功能障碍和奶牛妊娠， 在现场条件下由未经培训的人员进行。设想的系统是 一种易于使用，不需要提取或 色谱法，并将给出快速，定量的结果。定量 将用手动色度计完成， 直接连接到运行定制软件的电脑上该系统将建立在 在一个新的设备（患者待定），用于收集和处理 液体在几分钟内自动产生几毫升的 含有低分子化合物的澄清无蛋白质的滤液 重量.在第二阶段，将采用综合办法， 将开发试验运行的方式，将进行试验 在现场使用时是稳健的，因此修改后的 将寻求分析，其在奶牛场的用处将是 示范和原型系统能够吸引第三阶段的支持， 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.