量子点荧光探针的制备及其在金黄色葡萄球菌检测中的应用-微生物学专业论文.docxVIP

葡萄球菌的方法。在本检测体系中，羧基修饰的量子点(530nm)与 葡萄球菌的方法。在本检测体系中，羧基修饰的量子点(530nm)与 氨基修饰的DNA在EDC作用下脱水结合形成量子点核酸探针，再 互补结合BHQ(530nm)修饰的DNA，QD即发生荧光猝灭。当加入靶 序列后，靶序列通过更强的亲和力与QD-DNA探针杂交而BHQ与 QD分离，QD复合物的荧光复原。而加入误配碱基序列和其它菌的 DNA时，量子点的荧光强度不会复原。实验结果表明，通过检测QD 荧光的变化可对携带缸f基因的金黄色葡萄球菌进行定量检测，且此 检测方法具有高的灵敏性和特异性。该方法中目标DNA的浓度与量 子点复原的荧光强度与呈良好的线性关系，线性范围为0．2～1．2 IlM， 目标DNA的检测下限可达0．21．tM。 关键词：量子点，制备，荧光能量共振转移，金黄色葡萄球菌，tst AB AB STRACT Water-soluble quantum dot，a kind of semiconductor nanoparticles with excellent optical properties，Can be used as fluorescent probe or mark in the life science research．It has the prominent advantages such as broad excitation spectrum，symmetric emission spectrum and narrow full width of half maximum，good photochemical stability,continuous spectral band and the apparente anti-photobleaching compared to the traditionary fluorescent dye．In this paper,we synthesize CdSe／ZnS quantum dots in aqueous phase and apply it for detection of pathogen． Two major works were studied as follows： 1·Preparation of CdSe and CdSe／ZnS quantum dots in water phase． In order to solve the problems of toxic products，harsh reaction conditions and non water-solubility of quantum dots synthesis in organic solvent phase，we synthesized water-soluble CdSe quantum dots by using the NaHSe as precursor and mercaptoacetic acid as stabilizer．The CdSe／ZnS nuclear-shell quantum dots with higher quantum yield were synthesized based on CdSe quantum dots．In this preparation，the effects of stabilizer, pH and dosage to the fluorescent properties of quantum dots were discussed．Then the prepared coarse product was puri fied and characterized by fluorescence spectrum and transmission electron microscope．The results cl early showed that this method is simpl e and IIl intensity intensity is linear to target DNA concentration ranged from 0．2 to 1．2 pM． Keywords：Quantum dot，preparation，FRET，Staphylococcus a玎reus， oV 目 目 录 摘 要 ．． ．． ． ．．． ．．． ．．I ABSTRACT．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．III 第一章引言和文献综述．． ．．．