基于胶体金光热效应的快速检测方法研究及装置研制

发布时间：2018-05-20 14:43

本文选题：试纸条 + 胶体金　；参考：《山东师范大学》2017年硕士论文

【摘要】：近年来,食品安全问题越来越受到社会的重视,其中对食品中致病菌、农兽药残留等有害物质的检测方法研究是监督食品安全的重要保障。对食品中有害物质的检测方法有多种,胶体金(Gold nanoparticles,GNPs)试纸条检测方法是常用的快速检测方法之一。胶体金试纸条检测方法包括胶体金免疫层析技术和胶体金免疫渗滤技术,本论文主要研究基于胶体金免疫层析技术的检测方法。胶体金免疫层析技术是一种以GNPs作为显色物质的免疫层析检测技术,通常根据抗原上表位的数目分为双抗夹心法和竞争法,载体可以为硝酸纤维素膜(Nitrocellulose membrane,NC),玻璃等,广泛应用于食品、医疗检测等领域。传统胶体金免疫层析试纸条通常通过视觉检测法得出检测结果,但这种检测方法灵敏度一般较低。GNPs具有良好的光热效应,在532 nm激光照射下,GNPs分子被激发,产生大量热量。本论文制作了鼠伤寒沙门氏菌胶体金免疫层析式试纸条,在此基础上利用GNPs的光热性质,用热像仪进行光热检测,得出定量分析结果。由于热像仪价格相对仍较高,本论文进一步采用价格更低廉的红外温度传感器进行光热检测,建立了一种基于GNPs光热效应的层析式试纸条的传感器快速检测方法,研制了一种新型的光热检测装置。论文内容如下:第一章主要概述了试纸条检测方法的分类、GNPs的制备方法和性质,介绍了GNPs的标记技术和光热效应机理,重点介绍了胶体金试纸条的视觉检测方法和胶体金试纸条读卡器检测方法及其研究进展,以及基于GNPs光热效应的应用及其研究进展。第二章建立了一种基于GNPs光热效应的鼠伤寒沙门氏菌层析式试纸条检测方法。制备了GNPs,并与抗体组成金标抗体复合物,制作了鼠伤寒沙门氏菌免疫层析式试纸条,对其制作过程进行了优化。优化了光热检测的激光照射时间,分别用视觉检测法及热像仪光热效应检测法检测,得到了鼠伤寒沙门氏菌的最低检测限,分别为1.57×10~7 CFU·mL~(-1)和1.7×10~6 CFU·m L-1,光热效应检测法与视觉检测法相比,灵敏度提高了约9倍。第三章选用双抗夹心法商品化鼠伤寒沙门氏菌试纸条和竞争法商品化盐酸克伦特罗试纸条双抗夹心,利用视觉检测法及热像仪光热效应检测法进行了定量检测,确定了商品化胶体金试纸条采用视觉检测法及热像仪检测法检测鼠伤寒沙门氏菌及盐酸克伦特罗的最低检测限。鼠伤寒沙门氏菌试纸条视觉检测法和热像仪检测法的检测限分别为1.13×10~7 CFU·m L-1及1.2×10~6 CFU·mL~(-1),热像仪检测方法与视觉检测方法相比,灵敏度提高了约9倍。盐酸克伦特罗试纸条视觉检测法和热像仪检测法的检测限分别为5 ng·mL~(-1)及0.5ng·mL~(-1),热像仪检测方法与视觉检测方法相比,灵敏度提高了约10倍。第四章研制了一种小型红外温度传感器装置,将第二章和第三章中测试的自制和商品化鼠伤寒沙门氏菌试纸条和商品化盐酸克伦特罗试纸条用红外温度传感器进行光热检测。分别对激光照射距离、传感器测量距离及激光照射时间进行了优化,确定了胶体金免疫试纸条方法采用视觉检测法及红外传感器光热效应检测法检测鼠伤寒沙门氏菌及盐酸克伦特罗的最低检测限。自制鼠伤寒沙门氏菌试纸条视觉检测法和传感器检测法检测限分别为1.57×10~7 CFU·mL~(-1)及1.25×10~6 CFU·mL~(-1),传感器检测方法与视觉检测方法相比,灵敏度提高了约12倍。商品化鼠伤寒沙门氏菌试纸条视觉检测法和传感器检测法检测限分别为1.13×10~7 CFU·mL~(-1)及1.0×10~6 CFU·m L-1,传感器检测方法与视觉检测方法相比灵敏度提高了约11倍。盐酸克伦特罗试纸条视觉检测法和传感器检测法检测限分别为5 ng·m L-1及0.3 ng·mL~(-1),传感器检测方法与视觉检测法相比灵敏度提高了约16倍。第五章总结了本论文建立的基于GNPs光热效应的快速检测方法以及研制的检测装置,该方法能够实现待测物的快速检测,与试纸条视觉检测法相比灵敏度得到显著提高,且具有简单快捷、不需借助大型仪器及应用广泛的特点。
[Abstract]:In recent years, the problem of food safety has been paid more and more attention by the society. The research on detection methods of harmful substances such as pathogens in food, veterinary drug residues and other harmful substances is an important guarantee for supervision of food safety. There are many kinds of detection methods for harmful substances in food. The testing method of colloidal gold (Gold nanoparticles, GNPs) test strip is a common fast method. Colloidal gold immunochromatography and colloidal gold immunofiltration are one of the detection methods of colloidal gold test paper. This paper mainly studies the detection method based on colloidal gold immunochromatography. Colloidal gold immunochromatography is an immunochromatographic detection technique using GNPs as a chromogenic substance, usually based on the epitope of the antigen. The number is divided into double resistance sandwich method and competition method. The carrier can be widely used in food, medical inspection and other fields, such as Nitrocellulose membrane, NC, glass and so on. Traditional colloidal gold immunochromatography paper is usually obtained by visual detection, but the sensitivity of this detection method is generally lower than that of.GNPs. Good photothermal effect, under 532 nm laser irradiation, GNPs molecules are excited to produce a large amount of heat. In this paper, a colloidal gold immunochromatographic test paper for Salmonella typhimurium was made. Based on the photothermal properties of GNPs, the thermograph was used to detect the heat and heat, and the result of quantitative analysis was obtained. Further using the cheaper infrared temperature sensor to detect the light and heat, a fast detection method of the sensor based on the GNPs photothermal effect is established, and a new type of light and heat detection device is developed. The contents of the paper are as follows: the first chapter mainly outlines the classification of test paper detection method and the preparation method of GNPs. The marking technology and the photothermal effect mechanism of GNPs are introduced. The visual detection method of colloidal gold strip and the detection method of colloidal gold test paper reader and the research progress, and the application and research progress based on the GNPs photothermal effect are introduced. A kind of Salmonella typhimus based on the GNPs photothermal effect is established in the second chapter. The detection method of bacterial chromatography test paper strip was prepared. The GNPs was prepared and the antibody composition of the antibody was made up of the gold standard antibody complex. The preparation process of Salmonella typhimurium was made. The process of its production was optimized. The laser irradiation time of the photothermal detection was optimized. The mouse injury was detected by the visual detection method and the thermo thermal effect detection method respectively. The minimum detection limits for Salmonella Cambrian are 1.57 * 10~7 CFU. ML~ (-1) and 1.7 x 10~6 CFU. M L-1. The sensitivity of the photothermal effect detection is about 9 times higher than that of the visual inspection. The third chapter uses the double anti sandwich method to commercialize the Salmonella typhimurium test paper and the competitive method commercialized clenbuterol test paper double resistance sandwich. The quantitative detection method of visual detection and thermograph photothermal effect detection was carried out, and the minimum detection limit for the detection of Salmonella typhimurium and clenbuterol hydrochloride by visual detection and thermography was determined. The detection limits for the test paper and the thermograph test method of Salmonella typhimurium were respectively. For 1.13 * 10~7 CFU. M L-1 and 1.2 x 10~6 CFU. ML~ (-1), the sensitivity of the thermal imager is about 9 times higher than that of the visual detection method. The detection limit of the clenbuterol test paper and the thermal imager detection method is 5 ng mL~ (-1) and 0.5ng. A small infrared temperature sensor device was developed in Chapter fourth. The light and heat detection of the self-made and commercialized Salmonella typhimurium test paper and the commercialized clenbuterol test paper in the second and third chapters and the infrared temperature sensor for the commercialized clenbuterol test paper were tested in the second and third chapters. The distance of the laser irradiation, the measurement distance and the excitation of the sensor were measured respectively. The light irradiation time was optimized. The minimum detection limit for the detection of Salmonella typhimurium and clenbuterol was determined by the colloidal gold immunoassay strip method and the infrared sensor photothermal effect detection method. The test limits of the self-made test paper for Salmonella typhimurium and the detection method of sensor detection were 1.57 x 10~7 C, respectively. FU. ML~ (-1) and 1.25 x 10~6 CFU. ML~ (-1), the sensitivity of the sensor detection method is about 12 times higher than that of the visual detection method. The detection limits of the visual detection method and the sensor detection method for Salmonella typhimurium commercialization are 1.13 * 10~7 CFU mL~ (-1) and 1 x 10~6. The sensitivity increases about 11 times. The detection limits of the clenbuterol test method and the sensor detection method are 5 ng. M L-1 and 0.3 ng. ML~ (-1) respectively. The sensitivity of the sensor detection method is improved by about 16 times compared with the visual detection method. The fifth chapter summarizes the rapid detection method based on the photothermal effect based on GNPs in this paper and the fifth chapter. The developed detection device can realize the rapid detection of the object to be measured. Compared with the test paper vision detection method, the sensitivity is greatly improved, and it is simple and quick, without the need of large instruments and wide applications.
【学位授予单位】：山东师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TS207.3;O652

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