基于量子点标记的同步检测多血清型金葡菌肠毒素的免疫分析研究

In recent years, food safety has become one of the major concerns of our society. Particularly, food contaminations caused by pathogenic bacteria and the toxins pose a huge threat to public health. As a consequence, detection of bacterial toxins is a key link in the entire food safety system. Staphylococcal enterotoxins (SEs) are bacterial exotoxins secreted by staphylococcus aureus. SEs belong to a category potentialbiological weapon because of stable biologically active, so the highly sensitive and specific methods of detection of SEs are of critical importance in food safety and the war against terrorism. Different methods have been developed with the aim of detecting SEs, however, these assays are mostly used to detect the individual staphylococcal enterotoxin. Up to now, no immunoassay for screening multiple serotypes SEs using polyclonal or monoclonal antibodies has been reported. In this study, we will focus on the 'broad-spectrum' antibody production and 'multi-antibodies' for detection of multiple serotypes SEs, and establishing chemiluminescent immunoassay and immunochromatography based on quantum dots-labeling with highly sensitive, specific, and rapid, which will have potential applications for high-throughput screening and rapid detecting in public health. Meanwhile, docking techniquies will be tried to simulate the three-dimensional conformation between antigen and multi-antibodies to look for the rules of interaction between multi-antibodies and target molecules, which will provide scientific basis for applying multi-antibodies.

金黄色葡萄球菌肠毒素是引起食物中毒的常见生物毒素之一，也是一种重要的生物战剂，因而其检测一直备受重视。目前国内外尚无快速可靠同时筛查出多种血清型金黄色葡萄球菌肠毒素的免疫分析方法，其主要原因是"类特异性"广谱抗体难以获得、检测灵敏度低及假阳性问题。目前已有采用"复合抗原"和"复合抗体"技术制备广谱抗体的研究，但对于该类抗体应用于免疫分析的机理未见有报道，我们在大量前期研究的基础上，将从分子和空间结构等方面探索新型广谱抗体应用于免疫分析的机理，并通过引入纳米粒子提高检测系统的灵敏度，建立可以同时检测食品中痕量多血清型金葡菌肠毒素的化学发光免疫分析方法以及免疫层析检测方法，以期突破传统免疫分析中存在的广谱抗体制备困难、灵敏度低、基质影响严重等难题，为食品卫生领域中高通量的样品筛选和快速检测提供科学依据和技术支持。

金黄色葡萄球菌肠毒素是引起食物中毒的常见生物毒素之一，也是一种重要的生物战剂，因而其检测一直备受重视。目前对肠毒素的检测主要是ELISA和PCR法，国内外尚无快速可靠同时筛查出多种血清型金黄色葡萄球菌肠毒素的免疫分析方法，其主要原因是“类特异性”广谱抗体难以获得、检测灵敏度低及假阳性问题。目前已有采用“复合抗原”和“复合抗体”技术制备广谱抗体的研究，但对于该类抗体应用于免疫分析的机理未见有报道。.在本研究中我们通过单独免疫和复合免疫两种方式分别获得了特异性抗体和广谱抗体，并在此基础上配制了“复合抗体” 突破了传统免疫分析中存在的广谱抗体制备困难的问题。我们建立了三种形式的多种血清型金黄色葡萄球菌肠毒素的免疫分析方法：酶联免疫分析方法、胶体金免疫层析分析方法和量子点荧光免疫分析方法。并从分子和空间结构等方面初步探索了新型“复合抗体”应用于免疫分析的机理，以期为食品卫生领域中高通量的样品筛选和快速检测提供科学依据和技术支持。