COLORIMETRIC MEASUREMENT OF MILK PROGESTERONE

该 II 期 SBIR 申请提出了基于新颖的、 固相比色免疫分析系统，用于定量 牛奶中黄体酮的测定，其可行性 在第一阶段研究期间显示。浓度之间良好的相关性 采用常规方法测定牛奶中的孕酮 放射性标记示踪剂和新开发的酶免疫分析法 获得。该检测系统基于单克隆抗体的使用， 通过专有方法共价连接到微孔板孔中 与非放射性、基于酶的信号发生器相结合。阶段 II 研究的目的是将该检测方法开发成一种简单、可靠的检测方法 用于监测动情周期、产后卵巢周期的系统， 卵巢功能障碍和奶牛妊娠可以进行 由未经培训的人员在现场条件下进行。设想的系统是 一种易于使用、不需要提取或 色谱法，将给出快速、定量的结果。定量 将通过可耦合的手动色度计来完成 直接连接到运行自定义软件的计算机。系统将建成 一种用于收集和处理的新型设备（待患者使用） 几分钟内自动产生几毫升的液体 清澈、不含蛋白质的滤液含有低分子化合物 重量。在第二阶段，一种旨在简化 将开发测定运行的方式，将进行测定 适合在现场使用，进一步验证如此修改的 将寻求测定，其在奶牛场中的有用性将是 经证实，原型系统能够吸引 III 期支持 商业化将会得到发展。

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.