微体系HRP浓度与其催化化学发光衰减率线性关系研究  

Linear relationship between concentration of HRP and its chemoluminescence decays in the micro-channel system

作　　者：杨子岩[1] 梁平[2] 汪钦[2] 雷小英[2] 卢兹凡[2] 郭晏海[2] 向安[2] 

机构地区：[1]第四军医大学药学院学员二队,西安710032 [2]第四军医大学药学院药物基因组学教研室/全军基因诊断技术研究所,西安710032

出　　处：《现代仪器与医疗》2014年第1期1-4,12,共5页Modern Instruments & Medical Treatment

基　　金：国家"十一五""艾滋病和病毒性肝炎等重大传染病防治"科技重大专项支持(项目编号:2009ZX10004-311);国家自然科学基金项目(项目编号:81301513)

摘　　要：目的：了解微体系辣根过氧化物酶（HRP）催化化学发光与其浓度的相关性。方法：将HRP经核酸序列偶联于毛细管微腔内，通入微量反应底物进行化学发光检测；分析HRP催化化学发光反应与其浓度的相关性。结果：毛细管内HRP催化1．5μL底物的化学发光反应可分成急速衰减区（3．5×10^-4-1．3×10^4μg／μL）、衰减延缓区（9．7×10^-5-3．3×10^-5μg／μL）和发光平稳区（1．0×10^-5-6．7×10^-7μg／μL）；浓度为3．5×10^-4-6．7×10^6μg／μL的HRP催化化学发光衰减率RLus／T〉0且两者线性相关（R^2=0．967）。结论：分析微体系HRP浓度与化学发光衰减率（RLUs／T）的线性相关性，为微体系HRP化学发光定量检测生物分子提供新策略。Objective： Research the relationship between the concentration of horseradish peroxidase （HRP） and its chemiluminescence （CL） decays in micro-channel system. Method： Immobilize the HRP-streptavidin into a hydroxylation capillary, the micro-channel reaction system model in this study, with a single-stranded deoxyribonucleic acid （ssDNA） sequences that modify with biotin and amidogen. The relative luminescence units （RLUs） were being detected by a portable detector after injecting micro liter scale reaction substrates. Then the relationship between concentration of HRP and RLUs was been analysis through linear regression. Result： There were three RLUs characteristic regions, content rapidly decay in 3.5×10^-4-1.3×10^4μg/μL, slowly decay in 9.7×10^-5-3.3×10^-5μg/μL and steady region of 1.0×10^-5-6.7×10^-7μg/μL, basic on the rate of decay of RLUs for different HRP concentration. Most important, the rate of decay of RLUs （RLUs/T） was linearly related to the concentration of HRP （3.5×10^-4-6.7×10^6μg/μL, R2=0. 967）. Conclusion： The research of the linear relationship between concentration of HRP and RLUs/T afforded novel strategy for quantitative determination ofbiomolecules through chemiluminescent analysis in micro-channel system.

关 键 词：微反应体系 辣根过氧化物酶 化学发光衰减 线性相关性 

分 类 号：R331[医药卫生—人体生理学]