基于核酸适配体的化学发光检测腺苷  

Aptamer-based Chemiluminescent Detection of Adenosine

作　　者：鲁卢 严喜鸾[1] 肖义陂[2] 陈静雯[1] 肖鉴谋[1] 

机构地区：[1]南昌大学环境与化学工程学院,江西南昌330031 [2]南昌市洪都中医院,江西南昌330008

出　　处：《分析测试学报》2014年第1期73-77,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金青年科学基金资助项目(81102289);江西省自然科学基金资助项目(20132BAB205106);江西省教育厅青年科学基金资助项目(GJJ10053)

摘　　要：以羧基磁性微球为分离载体,固定氨基修饰的腺苷适配体,通过腺苷与生物素修饰的报告序列竞争结合氨基适配体,建立了一种化学发光检测生物小分子腺苷的方法.实验优化了磁性微球、氨基适配体、报告序列等参数,发现腺苷浓度在1.0 × 10-9～1.0×10-4 mol/L范围内与CL信号强度减少量呈良好线性关系,最低检出限达1.0×10-9mol/L,重复5次测定0.1 mmol/L腺苷的相对标准偏差为5.0％.方法成功用于实际尿样中腺苷的测定.该法可简单、灵敏、特异性地检测腺苷,有望用于临床诊断、药物分析等领域.Based on magnetic beads as separating carrier and adenosine as a specific competitor appeared,a method of aptamer-based chemiluminescence （CL） for the determination of adenosine was developed.Experimental parameters including magnetic beads,adenosine-aptamer and report DNA were optimized.Under the optimal conditions,the decreased CL intensity was proportional to the adenosine concentration in the range of 1.0 × 10-9-1.0 × 10-4 mot/L（r2 =0.99）,with a detection limit of 1.0 × 10-9 mol/L.The relative standard deviation （n =5） for 0.1 mmol/L adenosine is 5.0％.The present method was successfully applied in the determination of adenosine in real human urine.The method is simple,sensitive and specific for the detection of adenosine,and it is a promising approach for the determination of small biomolecules in the fields of clinical diagnosis,pharmaceutical analysis and so on.

关 键 词：化学发光 腺苷 适配体 磁性微球 辣根过氧化物酶 

分 类 号：O657.3[理学—分析化学] Q524[理学—化学]