基于纸芯片的盐酸克伦特罗和皮质醇检测

发布时间：2018-05-25 00:22

本文选题：纸芯片 + 盐酸克伦特罗　；参考：《大连理工大学》2015年硕士论文

【摘要】：生物标记物,是随着分子生物学和免疫学的研究深入,而发展起来的一类与细胞生长增殖有关的标志物。随着人们生活水平的提高和健康意识的增强,生物标记物的检测也变得愈发重要,传统的生物标记物的检测方法包括液质联用,气质联用等,这些方法大多需要专人操作大型仪器,而且花费高,耗时长。纸芯片作为一种新兴的检测平台具有质量轻、造价低、生物相容性好、无污染等很多优势,在生物标记物检测方面具有广阔的前景。本论文以盐酸克伦特罗和皮质醇为案例,结合免疫分析方法和高灵敏度的表面增强拉曼检测技术,开发了两种检测限低、实用性强的,以纸芯片为材质的生物标记物分析手段,具体内容包括：(1)基于并列开关型纸芯片和免疫增强拉曼的瘦肉精检测盐酸克伦特罗具有促进脂肪分解,提高动物体瘦肉率的作用,但是因其会对人体造成伤害而被我国明令禁止使用,为防止不法商贩为牟取暴利而滥用,盐酸克伦特罗的检测非常必要。本文提取了猪毛中的盐酸克伦特罗,采用喷蜡打印法制作了并列开关型纸芯片,合成了金纳米粒子,优化了金纳米粒子与信号分子MBA连接的反应时间和信号分子加入量,并在最佳条件下进行了盐酸克伦特罗抗体的连接,筛选了不同纸张上金标抗体的SERS增强效果,最终选择了whatman No.1滤纸作为芯片制作材料,在自主设计的纸芯片上实现了猪毛颗粒的过滤、富集,并最终通过开关控制流体流动进而实现了免疫反应,可以进行三种物质或者三个浓度物质的检测,检测限可以达到0.1 pg/mL。(2)基于折叠孔板型纸芯片和免疫增强拉曼的皮质醇检测皮质醇是评价机体应激程度的重要指标,其检测可以作为表征人们心理压力的客观依据,尤其是毛发中皮质醇检测更可以客观的反应人们长期的心理压力情况。本文考察了金标皮质醇抗体的修饰条件,筛选出的最佳滤纸whatman No.1滤纸为材质制作了折叠孔板型纸芯片,并利用纸芯片的折叠功能实现了人毛发提取液的过滤,竞争性免疫反应。该纸芯片还设计了96个并行单元,满足高通量检测要求,检测限可以达到0.1 pg/mL。最后,将折叠孔板型芯片用于便携拉曼光谱仪检测,检测限可以达到1pg/mL,为生物标记物的实时检测奠定了基础。
[Abstract]:Biomarkers are a kind of biomarkers related to cell growth and proliferation developed with the development of molecular biology and immunology. With the improvement of people's living standard and health awareness, the detection of biomarkers has become more and more important. Most of these methods require personnel to operate large-scale instruments, and expensive, time-consuming. As a new detection platform, paper chip has many advantages, such as light quality, low cost, good biocompatibility, no pollution and so on, so it has a broad prospect in biomarker detection. In this paper, clenbuterol hydrochloride and cortisol were taken as examples, combined with immunoassay and highly sensitive surface-enhanced Raman detection technique, two kinds of biomarkers with low detection limit and strong practicability were developed, which were based on paper chip. The specific contents include: 1) Detection of clenbuterol hydrochloride based on juxtaposition paper chip and immune-enhanced Raman has the effect of promoting fat decomposition and increasing lean meat rate of animals. However, it is forbidden to be used in our country because of its harm to human body. In order to prevent the illegal traders from abusing for profiteering, the detection of clenbuterol hydrochloride is very necessary. In this paper, clenbuterol hydrochloride was extracted from porcine hair, and the parallel switch paper chip was made by wax spray printing method. Gold nanoparticles were synthesized. The reaction time and the amount of signal molecule were optimized when gold nanoparticles were connected with signal molecule MBA. Under the best conditions, the clenbuterol hydrochloride antibody was connected, and the SERS enhancement effect of gold labeled antibody on different paper was screened. Finally, the whatman No.1 filter paper was selected as the chip material. On the independently designed paper chip, the pig hair particles can be filtered and enriched, and finally the flow of fluid can be controlled by switch, and then the immune reaction can be realized, which can be used for the detection of three substances or three concentration substances. The detection limit can reach 0.1 PG / m 路L ~ (2) cortisol detection based on fold-hole board chip and immuno-enhanced Raman is an important index to evaluate the stress degree of the body, and its detection can be used as an objective basis to characterize people's psychological stress. In particular, cortisol detection in hair can objectively reflect people's long-term psychological stress. In this paper, the modified conditions of gold-labeled cortisol antibody were investigated. The best filter paper whatman No.1 filter paper was used as material to fabricate the paper chip with folding hole, and the filter of human hair extract was realized by the folding function of the paper chip, and the competitive immune reaction was realized. The paper chip has also designed 96 parallel units to meet the high throughput detection requirements, and the detection limit can reach 0.1 PG / mL. Finally, the folded hole plate chip is used for the detection of portable Raman spectrometer. The detection limit can reach 1 PG / mL, which lays a foundation for the real-time detection of biomarkers.
【学位授予单位】：大连理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TS251.7;TN492

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