模拟酶化学发光信号放大与家禽细胞因子多组分免疫分析新方法研究

To meet the need of clinical detection in poultry industry and solve the problems such as low detection sensitivity, unable simultaneous multianalyte detection presented in assay method for poultry cytokines, this project involves in preparing and screening the monoclonal antibody of poultry cytokine, assembling high-density antibodies sensing array, synthesis of new-typed mimicking enzymes as well as its characteriztion of catalytic chemiluminescent property, construction of different mimicking enzymes signal amplification strategies, development of novel multianayte immunoassay method for poultry cytokines, and final realization of fast, wide linear range and automated jiont detection of poultry cytokine. Moreover, this project also study the combination of chemiluminescent imaging assay and flow-injection technique, establishment of new principle and method for ultrasensitive, high-througput chemiluminescent imaging immunoassay, and further interpreting the relationship between immune state in immune system of poultry organism and infection diseases. This project, creatively integrated merits of mimicking enzymes signal amplification and chemiluminescent imaging immunoassay, proposes a new ultrasensitive, high-throughput multianalyte immunoassay method, which will provide new standard and reasonable theoretic foundation for evaluating clinical immune response of poultry before and after vaccination and infection as well as have important academic significance and applied value

围绕当前家禽行业在临床检测方面的新需求，针对目前家禽细胞因子检测方法灵敏度低、无法多组分同时测定的问题，本项目拟制备、筛选家禽细胞因子单克隆抗体，组装高密度家禽细胞因子抗体传感阵列，合成人工模拟酶并表征其催化化学发光性能，设计模拟酶化学发光信号放大新策略，建立多种家禽细胞因子多组分免疫分析新方法，实现自动化、宽线性范围、快速的多种家禽细胞因子联合检测；并通过流动注射技术的联用构建自动化化学发光成像分析系统，发展超高灵敏度、高通量的化学发光成像免疫分析新方法与新原理，揭示家禽机体免疫系统的免疫状况及其与疾病之间的相互关系。本项目集模拟酶信号放大技术与化学发光成像免疫分析的优点，提出超高灵敏度、高通量的多组分免疫分析新方法，为临床家禽疫苗免疫和疾病感染前后的免疫应答水平提供全新的评价标准和健全的理论依据，具有重要的理论意义和应用价值。

当前家禽细胞因子检测方法存在灵敏度低、无法多组分同时测定等问题，无法满足家禽行业在临床检测方面的新需。本项目设计合成了多种类型的高性能人工模拟酶，用扫描电镜、透射电镜、X-射线衍射仪、紫外-可见光谱等手段表征了人工模拟酶的形貌、结构和模拟酶催化学发光性能；筛选了家禽细胞因子单克隆抗体，组装了高密度家禽细胞因子抗体传感阵列，设计了多种模拟酶化学发光信号放大新策略，建立了多种家禽细胞因子多组分免疫分析新方法，发展了超高灵敏度、高通量的化学发光成像免疫分析新方法与新原理，实现了自动化、宽线性范围、快速的多种家禽细胞因子联合检测，揭示了家禽机体免疫系统的免疫状况及其与疾病之间的相互关系，为临床家禽疫苗免疫和疾病感染前后的免疫应答水平提供全新的评价标准和健全的理论依据，具有重要的理论意义和应用价值。