基于漫反射光谱法的皮肤胆固醇检测技术研究

发布时间：2018-01-01 23:14

  本文关键词：基于漫反射光谱法的皮肤胆固醇检测技术研究　出处：《中国科学技术大学》2017年博士论文　论文类型：学位论文

  更多相关文章： 皮肤胆固醇 动脉粥样硬化性心血管疾病 漫反射光谱 漫反射吸光度

【摘要】：动脉粥样硬化性心血管疾病(atherosclerotic cardiovascular diseases,ASCVD)是危害人类健康的重大疾病之一,其诊断与预防已经逐渐引起社会的关注。皮肤胆固醇与ASCVD密切相关,成为ASCVD的新型生物标志物。采用漫反射光谱法测量受试者皮肤胆固醇水平,进而实现ASCVD的早期筛查和风险评估,具备无创、便捷、实时和原位等显著优点,有较好的发展前景。本文重点研究基于漫反射光谱法的皮肤胆固醇无创检测技术,包括研制皮肤胆固醇特异性识别和指示所需的试剂,设计集成化的皮肤胆固醇光谱检测系统。并开展实验研究,通过测量离体猪皮胆固醇和在体人体皮肤胆固醇来验证该方法和系统的可行性。本文的主要内容和取得的成果有:1、皮肤胆固醇特异性识别和指示所需的试剂研制以及光谱检测系统设计。分为两个部分:第一部分为基于"三滴法"皮肤胆固醇特异性识别和指示所需试剂的设计和研制,包括检测试剂"A-C-B"共聚物溶液和供氢体底物溶液。并通过检测离体猪皮内的胆固醇验证了试剂的有效性。第二部分为光谱检测系统的设计,分为硬件设计部分和软件设计部分。硬件部分包括光源单元、光学传输单元、信号探测单元、试剂反应腔单元等。软件部分编写了用于皮肤胆固醇光谱测量的控制分析软件。除此之外,还进行了壳体设计、附件设计等,最后完成整套系统的设计和研制。2、皮肤胆固醇光谱检测系统性能测试,测量影响因素研究和校正。首先,对设计的皮肤胆固醇光谱检测系统的性能进行测试,包括光源、传输光路及微型光纤光谱仪。采用标准漫反射板和与待测对象相似形态的去离子水验证系统的重复性。其次,对影响系统测量的因素进行了研究和优化,包括系统与组织接触压力的影响,辣根过氧化物酶(Horseradish Peroxidase,HRP)的活性,系统漂移和皮肤背景等。第三,采用研制的皮肤胆固醇光谱检测系统,采集漫反射光谱,求得漫反射吸光度A的曲线,确定特征波长和特征波段。最后,根据胆固醇与检测试剂"A-C-B"共聚物溶液以等分子比例结合的特性,采用梯度浓度的检测试剂模拟梯度浓度的皮肤胆固醇,计算漫反射吸光度A并与"A-C-B"共聚物溶液浓度进行相关性分析。结果表明,特征波长652 nm处的漫反射吸光度A和特征波段620-650 nm内的积分漫反射吸光度A与" A-C-B"共聚物溶液浓度成线性增长和显著正相关。3、采用皮肤胆固醇光谱检测系统实现离体猪皮胆固醇的测量。首先采用萃取法获得含有梯度浓度胆固醇的离体猪皮样本。确定了萃取的步骤和条件。然后采用光谱检测系统检测萃取后的样本内的胆固醇,并与萃取时间进行线性拟合分析。结果表明,采用特征波长652nm处的漫反射吸光度A和特征波段620-650 nm的积分漫反射吸光度A均可以区分和相对定量样本内的梯度浓度胆固醇,并与萃取时间成线性相关。4、采用皮肤胆固醇光谱检测系统实现人体在体皮肤胆固醇无创测量。征集受试者31名,采用设计的光谱检测系统采集受试者在体皮肤胆固醇,并获取受试者血液胆固醇。通过线性拟合和相关性分析受试者皮肤胆固醇与血液胆固醇中总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)和极低密度脂蛋白胆固醇(very low density lipoprotein cholesterol,VLDL-C)5项指标和由该5项指标衍生出的其它ASCVD风险指标的关系。结果显示:皮肤胆固醇与TC和LDL-C的变化趋势一致,并和二者显著正相关,控制年龄、性别和BMI等因素后,偏相关系数r分别为 0.813(P.01和 0.759(P＜0.001);皮肤胆固醇与 TG、HDL-C 和 VLDL-C 3项指标无显著性关系;皮肤胆固醇与非HDL-C、TC/HDL-C、LDL-C/HDL-C和AI四项指标之间均成线性关系,并成显著正相关,偏相关系数r分别为0.769(P0.01)、0.674(P0.01)、0.746(P0.01)和 0.674(P0.01);将皮肤胆固醇与任意两项参考指标进行组合分析,随着参考指标的增长,皮肤胆固醇也呈增长趋势。最后,基于上述参考指标在ASCVD风险评估中的参考标准将受试者划分为正常组、低风险组和高风险组,3组受试者之间的皮肤胆固醇具有显著性差异(单因素方差分析F值均大于1,P0.01),表明采用皮肤胆固醇能够实现不同ASCVD风险受试者的区分。综合上述研究表明,采用漫反射光谱法无创检测皮肤胆固醇将有助于ASCVD早期筛查和风险评估,对于该类疾病的防控具有一定的意义。
[Abstract]:Atherosclerotic cardiovascular disease (atherosclerotic cardiovascular, diseases, ASCVD) is one of the major diseases that endanger human health, its diagnosis and prevention has gradually attracted the attention of the society. The skin cholesterol is closely related with ASCVD, ASCVD has become a new biological marker. Measuring subjects skin cholesterol levels by using diffuse reflectance spectroscopy, and screening and risk assessment of early ASCVD, with noninvasive, convenient, real-time and in situ and other significant advantages, it has good prospects for development. This article focuses on the study of skin cholesterol diffuse reflectance spectroscopy noninvasive detection technology, including the development of skin cholesterol reagent specific recognition and instructions required for the skin cholesterol spectrum detection system integrated design and the experimental study was carried out by measuring the in vitro and in vivo human skin skin cholesterol cholesterol to verify the method and system The feasibility. The main contents of this paper are as follows: 1. Design and development of reagent spectrum detection system of skin cholesterol specific recognition and instructions required. Divided into two parts: the first part is based on the "three drops" skin cholesterol specific recognition and instructions required to test the design and development of agent, including kit "A-C-B" copolymer solution and hydrogen donor substrate solution. And through the detection of cholesterol in vitro pig validated reagents. The second part is the design of spectrum detection system, divided into hardware design and software design. The hardware part includes a light source unit, optical transmission unit, a signal detection unit, reagent the reaction cavity unit. The software is used to control skin cholesterol spectrum analysis software. In addition, also carried out the shell design, accessories design, finally completed the whole design of system With the development of.2, skin cholesterol spectrum detection system performance test, and the correction factor measurement. Firstly, the performance of skin cholesterol spectrum detection system design for testing, including the light source, optical transmission and micro fiber spectrometer. The repeatability standard diffuser and deionized water with the measured object similarity form verification system. Secondly, the research and optimization of factors affecting the measurement of the influence, including system and organization of the contact pressure, horseradish peroxidase (Horseradish Peroxidase, HRP) activity, system drift and skin background. Third, the skin cholesterol spectrum detection system development, acquisition by diffuse reflectance spectroscopy, diffuse reflectance the absorbance of A curve, determine the characteristics of wavelength and spectral characteristics. Finally, according to the characteristics of cholesterol and the kit "A-C-B" copolymer solution with molecular ratio with the harvest Simulation of skin cholesterol concentration gradient detection reagent concentration gradient calculation, diffuse reflectance absorbance A and analyzed the correlation with the concentration of A-C-B copolymer solution. The results show that the integral features of diffuse diffuse reflection at the wavelength of 652 nm A and 620-650 nm absorbance characteristic band in reflection absorbance A and "A-C-B" copolymer concentration linear growth and significant positive correlation with.3, skin cholesterol spectrum detection system achieve measurement of body cholesterol. The pigskin extraction method with gradient concentration of cholesterol in vitro. The process and conditions of pigskin sample extraction is determined. Then the spectrum detection system after extraction of cholesterol in the sample, and the linear regression analysis and the extraction time. The results show that the integral absorbance of A using diffuse reflectance characteristics of diffuse reflectance at a wavelength of 652nm absorbance of A and 620-650 nm are the characteristic bands To distinguish between the concentration gradient and relative quantitative cholesterol within the sample, and a linear relationship between the.4 and the extraction time, the skin cholesterol spectrum detection system to achieve human skin in vivo noninvasive measurement of cholesterol. For 31 participants, the system design of the spectra of subjects in skin cholesterol, and obtain the subjects blood cholesterol. Through the analysis of linear fitting and correlation between the subjects of skin cholesterol and blood cholesterol and total cholesterol (total cholesterol TC), triglyceride (triglyceride, TG), high density lipoprotein cholesterol (high density lipoprotein cholesterol, HDL-C), low density lipoprotein cholesterol (low density lipoprotein cholesterol, LDL-C) and very low density lipoprotein cholesterol (very low density lipoprotein cholesterol, VLDL-C) between the 5 indexes and derived from the 5 indicators of the ASCVD risk index. The results showed that the change trend of skin cholesterol and TC and LDL-C, significant positive correlation, and the two control factors such as age, gender and BMI, the partial correlation coefficient r was 0.813 (P.01 and 0.759 (P < 0.001); skin cholesterol and TG, there was no significant relationship between the 3 indexes and HDL-C VLDL-C; skin cholesterol and non HDL-C, TC/HDL-C, as are the linear relationship between the four indexes of LDL-C/HDL-C and AI, and a significant positive correlation, partial correlation coefficient r was 0.769 (P0.01), 0.674 (P0.01), 0.746 (P0.01) and 0.674 (P0.01); the two reference index of skin cholesterol with any combination of analysis, with the reference index of the growth of skin cholesterol showed a growth trend. Finally, the reference standard reference index in ASCVD risk evaluation based on the subjects were divided into normal group, low risk group and high risk group, the skin cholesterol between the 3 groups of subjects with significant Difference (single factor analysis of variance F-measure was greater than 1, P0.01), show that the skin cholesterol can distinguish different ASCVD risk subjects. The research shows that using diffuse reflectance spectroscopy in noninvasive detection of skin cholesterol will help ASCVD early screening and risk assessment, and has a certain significance for prevention and control this kind of disease.

【学位授予单位】：中国科学技术大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R54;O657.3

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