基于金磁微粒的人血清总睾酮化学发光检测方法的建立

发布时间：2018-04-06 06:06

本文选题：GoldMag~R金磁微粒　切入点：睾酮　出处：《西北大学》2014年硕士论文

【摘要】：睾酮(Testosterone, T)作为重要的性激素之一,其水平影响着机体的健康状况,对其含量的准确检测有助于临床相关疾病的辅助诊断。睾酮为小分子物质,具有不易从结合蛋白解离的特性,加之血液中存在大量与其具有相似抗原活性位点的结构类似物或内源性物质,为其准确测定带来难度。本团队具有自主知识产权的GoldMag金磁微粒,因其具有较大的比表面积可以偶联足够的蛋白量,良好的超顺磁性保证其在外磁场中的分离操作,可实现高检测灵敏度和宽线性范围的检测需求,因而被用于检测血清样本微量物质如睾酮激素等的检测方法的建立。本研究以GoldMag金磁微粒作为睾酮抗原的固相载体,以辣根过氧化物酶(HRP)-鲁米诺(luminol)-过氧化氢(H2O2)化学发光系统作为检测体系。采用间接竞争法,建立了具有高灵敏度、特异性和重现性好,方便快捷的磁酶免化学发光法,经过评价方法实现了对临床样本人血清中总睾酮含量的测定。对体系免疫磁珠制备条件、免疫反应条件、抗原抗体用量、发光反应条件等进行研究和优化,最终建立体系的线性范围为0.05-5ng/ml,线性相关系数R20.99。方法学性能评估显示最低检测限为0.038ng/ml。用该方法与商业化的LIASON睾酮定量测定试剂盒(化学发光法)对临床120例健康人血清总睾酮含量进行检测并对比,两种方法的相关系数为0.97587,显示两种方法具有很好的相关性,表明建立的检测方法对临床样本人血清总睾酮的测定具有较高的准确度。
[Abstract]:As one of the important sex hormones, the level of testosterone Testosterone (T) affects the health status of the body, and the accurate detection of its content is helpful to the auxiliary diagnosis of clinically-related diseases.Testosterone is a small molecular substance, which is difficult to dissociate from binding proteins. In addition, there are a large number of structural analogs or endogenous substances with similar antigenic active sites in the blood, which makes it difficult to determine accurately.The team has its own intellectual property GoldMag gold particles, because of its large specific surface area can be coupled enough protein, good superparamagnetic to ensure its separation in the external magnetic field,It can achieve high detection sensitivity and wide linear range of detection requirements, so it is used for the detection of trace substances in serum samples, such as testosterone.In this study, GoldMag gold particles were used as solid carrier of testosterone antigen, and HRP- luminoline-H _ 2O _ 2 chemiluminescence system of horseradish peroxidase (horseradish peroxidase) was used as detection system.By using indirect competition law, a high sensitivity, specificity, reproducibility, convenient and rapid magnetoenzyme immunochemiluminescence method was established. The total testosterone content in human serum was determined by the evaluation method.The preparation conditions of immunomagnetic beads, the conditions of immune reaction, the dosage of antigen and antibody and the conditions of luminescence reaction were studied and optimized. The linear range of the system was 0.05-5 ng / ml, and the linear correlation coefficient was R20.99.Methodological performance evaluation showed a minimum detection limit of 0.038 ng / ml.This method was compared with the commercial LIASON testosterone quantitative assay kit (chemiluminescence method) to detect and compare the serum total testosterone content of 120 healthy people. The correlation coefficient of the two methods was 0.97587, which indicated that the two methods had good correlation.The results showed that the established method was accurate for the determination of serum total testosterone in clinical samples.
【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R446.11

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