基于共振瑞利散射血清蛋白测试技术及POCT仪器的研究

发布时间：2018-07-17 02:57

【摘要】：血清中蛋白质浓度在临床中有重要诊断意义，在蛋白质分析领域，广泛应用紫外可见分光光度计、荧光分光光度计等精密仪器，这些仪器适合医院化验室使用，，但由于它们的复杂、昂贵、体积庞大、耗电量高等因素，而无法满足现场检测POCT (Point of CareTesting)的要求。POCT是临床诊断技术中发展的趋势之一，它使用的仪器操作简便、工作稳定、适于携带。针对这种迫切需要，本文提出了一种基于共振瑞利散射（RRS）适合现场检测的血清蛋白质测试新方法。论文介绍了RRS国内外的研究现状、基本概念和基本理论；采用荧光分光光度计和紫外－可见分光光度计研究了四羧基酞菁锌－蛋白质和四氨基酞菁铜体系的共振光散射光谱和吸收光谱进行，在pH 3.6的Britton－Robinson（B－R）缓冲溶液中，蛋白质与四羧基酞菁锌发生相互作用，使体系在波长475nm处产生共振散射增强；进一步分析和实验表明：该溶液强吸收波长为420nm附近的蓝紫光波段，在该激励光作用下，其共振波长处会产生RRS，在一定蛋白质浓度范围内溶液的RRS强度与蛋白质的含量成比例，可以利用四羧基酞菁锌为光谱探针的共振散射法来测定血清蛋白，其线性检出范围为10～50mg/mL，检出限为0.001mg/mL；在此基础上，确定了主体以405nm宽禁带半导体激光器为激励光源，以475nm窄带带通滤光片为单色器，以基于蓝光增强光敏二极管的低噪声高增益光电放大器为探测器的血清蛋白质POCT测试装置的总体方案。论文所做工作还包括：对金属酞菁配合物的合成工艺和它们红外光谱表征的研究，采用4－羧基邻苯二甲酸酐和尿素为原料，钼酸铵为催化剂，固相法合成四羧基酞菁锌和四氨基酞菁铜，并通过红外光谱的表征得到证实；体系的稳定性、酸度对体系的影响、缓冲溶液用量、离子强度、干扰物质等影响检测灵敏度因素的讨论；血清蛋白质POCT测试装置样机的研制，并用它来对血清蛋白质进行测试。实验结果表明：新开发的血清蛋白质测试装置具有体积小、成本低、功耗小、使用方便等一系列优点，改进后可望在现场检测得到广阔的应用。
[Abstract]:The concentration of protein in serum is of great significance in clinical diagnosis. In the field of protein analysis, precision instruments such as UV-Vis spectrophotometer and fluorescence spectrophotometer are widely used, which are suitable for use in hospital laboratories. However, because of their complexity, high cost, huge volume, high power consumption and other factors, they can not meet the requirements of field detection of POCT (Point of career testing). POCT is one of the developing trends in clinical diagnosis technology. Suitable for carrying. To meet this urgent need, a new method for testing serum proteins based on resonance Rayleigh scattering (RRS) is proposed. This paper introduces the research status, basic concepts and basic theories of RRS at home and abroad. The resonance light scattering (RLS) spectra and absorption spectra of the system of zinc tetracarboxylphthalocyanine zinc protein and copper tetra aminophthalocyanine were studied by fluorescence spectrophotometer and UV-Vis spectrophotometer in the Britton-Robinson (B-R) buffer solution at pH 3.6. The protein interacts with zinc tetracarboxylphthalocyanine, which results in enhanced resonance scattering at the wavelength of 475nm. Further analysis and experiments show that the strong absorption wavelength of the solution is the blue violet band near 420nm. The RRS is produced at the resonance wavelength. The RRS intensity of the solution is proportional to the content of the protein in a certain range of protein concentration. The RRS intensity can be determined by the resonance scattering method using zinc tetracarboxylphthalocyanine as the spectral probe. The linear detection range is 10 ~ 50 mg 路mL ~ (-1) and the detection limit is 0.001 mg / mL. On the basis of this, it is determined that the main body is 405nm wide band gap semiconductor laser as the excitation light source, and the 475nm narrow band pass filter is used as the Monochromator, the linear detection range is 10 ~ 50 mg 路mL ~ (-1) and the detection limit is 0.001 mg 路mL ~ (-1). The overall scheme of a low noise and high gain photoelectric amplifier based on blue light-enhanced Guang Min diode as detector for measuring serum protein. The work of this paper also includes: the synthesis of metal phthalocyanine complexes and their infrared spectrum characterization, using 4-carboxyphthalic anhydride and urea as raw materials, ammonium molybdate as catalyst, Zinc tetracarboxylphthalocyanine and copper tetracarboxylphthalocyanine were synthesized by solid phase method, and characterized by IR spectra, the stability of the system, the effect of acidity on the system, the amount of buffer solution, the ionic strength, The factors affecting detection sensitivity, such as interfering substances, were discussed, and the prototype of serum protein POCT testing device was developed, and it was used to test serum protein. The experimental results show that the newly developed serum protein testing device has a series of advantages, such as small volume, low cost, low power consumption, easy to use and so on. The improved device is expected to be widely used in the field detection.
【学位授予单位】：中北大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：TH776;R446.11

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