基于图像传感器的金标免疫分析技术研究
文献类型：学位论文
| 作者 | 陈贝特 |
| 学位类别 | 硕士 |
| 答辩日期 | 2013 |
| 授予单位 | 中国科学院上海光学精密机械研究所 |
| 导师 | 黄立华 |
| 关键词 | 光电检测 光学生物传感器 金标免疫层析检测 图像传感器 POCT |
| 其他题名 | Research of Gold Labeled Immunoassay Technology based on the Image Sensor |
| 中文摘要 | 免疫测定是基础研究和临床检测中广泛使用的一种技术，遍及医学检测的各个领域。随着免疫测定技术的不断发展，许多新技术取代了传统的检测方法，金标免疫层析检测法，亦称金标免疫层析试纸条检测便是其中之一。金标免疫层析试纸条检测具有诸多优点，包括检测速度快、操作简便、无污染、试剂稳定等，已受到临床检验界的极大关注，并发展成为临床检验的前沿领域。 POCT (Point of Care Testing)以其方便、快速、高效而受到了人们的青睐，近年在国内外都得到了快速发展。随着中国医改的实施以及防病健康知识的提高，今后POCT在健康保健、疾病预防监控中的需求量更多，发挥的作用更大。 但是，目前国内外金标免疫层析试纸条(简称：金标试纸条)定量检测的POCT仪器的性能并不理想，并且国外进口仪器价格较高，使该类型仪器在国内的应用受到了限制。在已经出现的金标试纸条检测POCT仪器中，其测量方式主要分为扫描测量和成像测量。对于采用扫描测量方式对金标试纸条进行检测的系统，其机械结构复杂，测量时间相对较长，且易受外界干扰。成像测量较扫描测量，具有快速、稳定等优点。但是对于采用成像测量方式的检测仪器，我们尚未看到对于照明和图像质量进行相关研究的检测仪器，而恰恰这两个方面会直接影响到仪器的检测性能。因此，研究并发展具有中国自主知识产权的高性能金标试纸条定量检测的POCT仪器的具有重要的意义与价值。 围绕着研制出一种具有高灵敏度、高重复性、快速、稳定、便携、功耗低、成本低、结构简单等优点的成像型金标试纸条定量检测POCT仪器的目标，本论文的主要内容如下： (1) 概述了POCT的发展及应用、金标免疫层析试纸条POCT定量检测仪器的发展现状、图像传感器以及DSP技术的发展状况，为本论文工作的开展奠定了必要的基础。 (2) 深入研究了金标试纸条定量检测理论，分析了纳米金颗粒的光学特性以及胶体金免疫标记技术的基本原理。之后，着重研究了金标试纸条结构，以及与本课题有关的双抗夹心法金标免疫层析反应原理。 (3) 开展了基于图像传感器的成像型金标试纸条检测POCT仪器(简称：金标条阅读仪)的整体设计。本论文从光学模块、控制系统模块和图像数据算法模块这三个模块开展金标条阅读仪的设计。同时，根据成像型仪器的特点，提出了提升仪器性能的关键技术点，即优化设计金标试纸条上照明均匀性和图像传感器成像参数，以此来提高金标条阅读仪的检测灵敏度。最后，详细介绍了金标条阅读仪设计的主要组成：OV663图像传感器、TMS320C6747 DSP芯片和嵌入式实时操作系统DSP/BIOS。 (4) 开展了金标条阅读仪的详细设计，具体内容包括金标条阅读仪照明均匀性设计，OV6630图像传感器成像参数优化设置，仪器内部图像硬件设备操做方法和电路连接，C6747 DSP芯片内部相关模块的软件设计，定量检测的图像数据处理算法设计，以及上述软件设计的操作流程。针对仪器的成像控制参数优化设计，提出了评价图像质量的两个评价参数。在上述详细设计的基础上，研制了一台金标条阅读仪样机，并介绍了该样机的工作过程。 (5) 开展了金标条阅读仪性能评价的实验。使用所研制的金标条阅读仪对系列浓度的心肌肌钙蛋白I(cTnI)金标试纸条进行检测实验研究。仪器对高浓度cTnI金标试纸条(64 ng/ml)测量结果的变异系数仅为0.134%，对低浓度cTnI金标试纸条(0.25 ng/ml)测量结果的变异系数仅为2.790%；检测灵敏度优于0.25 ng/ml；并采用4-PL曲线对实验测量的数据进行拟合，拟合相关系数R^2优于0.99；完成一次检测时间约为1秒。实验数据表明，该金标条阅读仪具有重复性好、灵敏度高、动态范围大、响应特性好、检测速度快等优点。 |
| 英文摘要 | Immunoassy is a sensitive detection technique in the field of clinical diagnostics, especially in clinical detection and medical studies. With the development of immunoassy, traditional detection methods in clinical diagnostics have been replaced by many new techniques. One of the new techniques is the gold immunochromato- graphic assay (GICA). With the advantages of high efficiency of detection, easy of operation, pllution free and reagent stable, GICA has gained great attention in clinical diagnostics. Because of its characteristics of convenient, fast and efficient, POCT (Point of Care Testing) has attracted much research interest and been rapid developed at home and abroad in recent years. With the implementation of China''s medical reform and the improvement of knowledge of disease prevention and health, POCT will play an increasingly important role in health care, disease prevention and monitoring in the future. However, the performance of POCT instrument for colloidal gold lateral flow (LF) strip quantitative detection is low, and the price of imported POCT instrument is expensive. There are mainly two forms of measurements, i.e. scanning measurement and imaging measurement. For the scanning measurement, the measurement reproducibility would be affected by instrument vibration and other disturbing factors, and the measurement time is relatively long. Additionally, the scanning mechanism also complicates the instrument’s structure. As for the imaging measurement, the LF strip picture is captured by image sensor. And image processing methods are used for quantitative analysis of the LF strip. So the imaging measurement is quick, stable, and has simple mechanical structure, etc. However, the image quality of the LF strip would directly affect the detection performance of the instrument. For example, illumination uniformity on the LF strip and the control parameters of image sensor has great impact on the image quality, and thus affect the measurement results. But the reported imaging measurement-based instruments are rarely concerned about the design of illumination uniformity and the control parameters.This situiation indicates that the research and development of high performance POCT instrument for colloidal gold LF strip quantitative detection with Chinese independent intellectual property rights is of great significance and value. Aiming at researching and designing POCT instrument based on image sensor for colloidal gold LF strip quantitative detection with properties of sensitive, reproducible, fast, stable, portable, low power consumption, low cost, simple structure, etc., the main research work in this paper is as follows: (1) An overview of the development and application of POCT and of the development of DSP technology, as well as of the development of POCT instrument for GICA quantitative detection is introduced. (2) The GICA test strip quantitative detection theory, the optical properties of gold nanoparticles and the basic principles of immunogold labeling technique are analysed. Then deep researches on the colloidal gold LF strip structure and on the principle of the double-antibody sandwich GICA reaction have been done. (3) The overall composition of the colloidal gold LF strip reader based on image sensor is researched and designed. The basic design modules of colloidal gold LF strip reader are introduced in three modules, i.e. optical module, control module and image algorithm module. Meanwhile, a new way to improve the performance of colloidal gold LF strip reader by the design of illumination uniformity and the optimization of image sensor’s image control parameters is proposed. Then, the basic design resources are introduced, including OV6630 image sensor, TMS320C6747 DSP chip and the embed real time operation system DSP/BIOS. (4) The colloidal gold LF strip reader is researched and designed, including illumination uniformity design, image control parameters settings design of OV6630, internal hardwares operating methods and circuit connections, the software operation of C6747 DSP chip internal modules and the image data processing algorithms of quantitative detection. Two evaluation parameters are defined to evaluate the image for the optimization of image control parameters. At last, a prototype of colloidal gold LF strip reader has been developed on the basis of the detailed design, and the working process of this reader is introduced. (5) Colloidal gold LF strips of Cardiac troponin I (cTnI) are tested by the colloidal gold LF strip reader. The coefficients of variation of 20 times repetitive experiments with 0.25 ng/ml and 64 ng/ml are respectively 2.790% and 0.134%. The functional sensitivity is better than 0.25 ng/ml. The test results are fitted by 4-PL curve model, whose R^2 is more than 0.99, and the single measurement time is less than 1 sencond. It proofs that the design of colloidal gold LF strip reader is of properties of high sensitivity, wide dynamic range, good response property and reproducibility. |
| 语种 | 中文 |
| 源URL | [http://ir.siom.ac.cn/handle/181231/16779]  |
| 专题 | 上海光学精密机械研究所_学位论文 |
| 推荐引用方式 GB/T 7714 | 陈贝特. 基于图像传感器的金标免疫分析技术研究[D]. 中国科学院上海光学精密机械研究所. 2013. |
入库方式： OAI收割
来源：上海光学精密机械研究所
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