CHEMILUMINESCENT WESTERN BLOT SYSTEM FOR AIDS DIAGNOSIS

这项提议的第一阶段的目标是建立一个化学发光 基于化学发光的艾滋病免疫印迹检测系统 碱性磷酸酶底物AMPPD(商标)。的优势之一 该系统将提高灵敏度，超过现有的西方斑点 测试，这会导致染色的膜条。另外，一份永久的记录 结果将直接在胶片上产生。一种新型的蛋白质印迹系统 使用牛血清白蛋白(BSA)和商用抗体将 用于开发一种新的发光放大系统，该系统将被 硝化纤维素膜的必需品。阻塞条件和灵敏度 也将得到优化。发光放大系统将导致 化学发光发射，最大发射波长约为540 nm。这 波长是捕捉化学发光信号的最佳匹配 宝丽来瞬间黑白胶片。发展出的最佳条件 有了BSA模型，系统将应用于杜邦的组件 HIV Western印迹试剂盒，使用该试剂盒的非感染性对照来测试 化学发光系统的性能和灵敏度。结果是 在即刻胶片上获得的将适用于

The goal of Phase I of this proposal is to establish a chemiluminescent detection system for AIDS Western blotting based on the chemiluminescent substrate for alkaline phosphatase, AMPPD (trademark). One advantage of this system will be increased sensitivity, over existing Western blots tests, which result in stained membrane strips. Also, a permanent record of results will be produced directly on film. A model Western blot system using bovine serum albumin (BSA) and commercially available antibodies will be used to develop a new luminescence amplifying system which will be necessary for nitrocellulose membrane. Blocking conditions and sensitivity will also be optimized. The luminescent amplifying system will result in a chemiluminescent emission with a maximum at approximately 540 nm. This wavelength is a good match for capturing the chemiluminescent signal on Polaroid instant black and white film. The optimal conditions developed with the BSA model system will then be applied to components of a DuPont HIV Western blot kit, using the kit's noninfectious controls to test the performance and sensitivity of the chemiluminescent system. The results obtained on instant film will be suitable for