【作者】 石晓彤；

【导师】 殷学锋； 方群；

【作者基本信息】 浙江大学 ， 分析化学， 2010， 博士

【摘要】 免疫分析是利用抗原和抗体之间的特异性相互作用对待测物进行检测的一种方法,具有很高的特异性和灵敏度,在临床检验、生化分析、环境监测、食品安全等领域的应用十分广泛。其中,非均相免疫分析,尤其是酶联免疫吸附测定法(Enzyme-linked immunosorbent assay,ELISA),是当前临床及实验室医学领域进行定量检测占主导地位的分析技术。然而,常规的免疫分析方法,特别是ELISA,孵育时间长,多采用手工操作,分析过程繁琐费时,试样和试剂的消耗量过大,限制了其进一步发展和实际应用。微流控学是上世纪九十年代发展起来的在微米级结构中操控纳升至皮升级流体的科学和技术。微流控分析系统由于其独特的微尺度效应,具有反应时间短、分析速度快、试剂和样品消耗少、易集成化和自动化等优点。将微流控分析技术与免疫分析结合,在微流控芯片的微通道内进行免疫分析操作,能有效降低系统的试样试剂消耗、提高分析速度和通量、实现分析系统的集成化和自动化。第一章对各种微流控免疫分析技术及其研究进展进行了综述,尤其对微流控非均相免疫分析方法及自动化的微流控非均相免疫分析系统的研究进展进行了重点介绍。第二章发展了一种基于毛细管和缺口管阵列技术的微流控非均相免疫分析系统。以缺口管阵列作为试样和试剂储液池,以毛细管阵列作为取样探针和免疫反应通道,以重力作为驱动力,采用直线滑台自动控制缺口管阵列的移动,实现多通道免疫反应操作的完全自动化。该系统被成功地应用于人IgG分析,在10min内同时完成了对9个IgG样品的同步分析。系统对IgG的检测限为1.0μg/mL,分析重现性为1.8%(RSD,n=9),每次测定试剂和试样消耗为1.1μL。该系统具有结构简单、液体操控自动化、试样和试剂消耗低、易于实现阵列化的特点,为临床检验分析的自动化及高通量化提供了一种新途径。第三章发展了一种基于缺口管阵列和微蠕动泵的聚二甲基硅氧烷(PDMS)免疫分析芯片。其中,建立了一个基于微型齿轮蠕动泵的液流驱动系统,具有结构简单、容易搭建、驱动可靠、流量可调(0.21-4.4μL/min)的优点。将蠕动泵与缺口管阵列配合,初步在PDMS芯片上实现多通道的人IgG的非均相免疫分析。系统对试样及试剂的消耗约0.4μL,对IgG的检测限为1.0μg/mL,可对5个样品进行同步分析,总分析时间约8 min。更多还原

【Abstract】 Immunoassay is a well-established analysis technique with high specificity and sensitivity based on specific interaction between antigen and antibody,and has been widely used in clinical diagnosis,biochemical analysis,environment monitoring and food safety control.Heterogeneous immunoassay,especially enzyme-linked immunosorbent assay(ELISA),is the most important and widely-used quantitative method in the field of clinical medicine.However,in most of the traditional immunoassays,the operations are usually carried out manually,which are tedious and time-consuming,and the consumption for sample and reagent is among several hundreds of microliters.Microfluidics is the science and technology that manipulate small(nL-pL scale) amount of fluids inμm-scale channels and microfluidic systems.Microfluidic analytical systems have the advantages of fast analysis,low sample/reagent consumption and potential for integration and automation.The combination of microfluidic technique with immunoassay provides significant improvements in analysis speed and throughput,sample/reagent consumption,and integration and automation of the system.In chapter 1,the progress in microfluidic immunoassay systems,especially in automated heterogeneous immunoassay,is reviewed.In chapter 2,an automated microfluidic immunoassay system was developed based on capillary and slotted-vial arrays,in which fully automated operation for multi-step immunoassay was achieved.The system was applied in the immunoassay of human IgG.The assay for nine human IgG samples was achieved within 10 min. The limit of detection for IgG was 1.0μg/mL.The precision of the system was 1.8% (RSD,n=9).The sample and reagent consumption was 1.1μL for each cycle.In chapter 3,a microfluidic immunoassay system was developed based on slotted-vial array,microperistaltic pump and PDMS chip.The peristaltic pump with a micro-gear was used for liquid driving in six microchannels in PDMS chip,which had advantages of simple structure,ease of building,good reliability and adjustable flow rate range from 0.21μL/min to 4.4μL/min.The feasibility of the system was demonstrated in a preliminary immunoassay for human IgG.The assay for five IgG samples was achieved within 8 min.The sample/reagent consumption was only 0.4μL,and the limit of detection was 1.0μg/mL IgG.更多还原