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基于适配体的胶体金色谱层析试纸条快速检测肌酸激酶同工酶

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基于适配体的胶体金色谱层析试纸条快速检测肌酸激酶同工酶
Aptamer-Based Colloidal Gold Chromatography Strips for Rapid Detection of Creatine Kinase-MB
投稿时间:2018-10-25
DOI:10.15918/j.tbit1001-0645.2019.s1.020
中文关键词:适配体肌酸激酶同工酶层析试纸心肌和肌肉损伤长期飞行
English Keywords:aptamerCreatine-Kinase MBchromatography stripsmyocardial and muscle injurylong-duration space missons
基金项目:国家重点科学仪器设备开发专项课题资助(2012YQ04014005)
作者单位
李永瑞北京理工大学 生命学院, 北京 100081
姚梦迪北京理工大学 生命学院, 北京 100081
冯薇北京理工大学 生命学院, 北京 100081
吕雪飞北京理工大学 生命学院, 北京 100081
邓玉林北京理工大学 生命学院, 北京 100081
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中文摘要:
研究基于适配体的胶体金色谱层析试纸条快速检测肌酸激酶同工酶,用于揭示长期飞行下航天员潜在的心肌和肌肉损伤.利用SELEX方法筛选得到的肌酸激酶同工酶适配体,并优选出与肌酸激酶同工酶有较强亲和能力的核酸适配体应用于胶体金免疫层析试纸条,通过优化胶体金颗粒大小、缓冲液浓度等条件,成功建立了一种基于适配体的胶体金试纸法用于CK-MB的快速检测.实验结果表明,通过基于磁珠的核酸适配体筛选方法筛选出的与肌酸激酶同工酶有亲和力的核酸适配体,能够用于构建适配体-适配体夹心胶体金免疫层析试纸条并成功应用于82 nM以上的肌酸激酶同工酶的快速、可视化检测,同时,该免疫层析试纸条具有良好的特异性和稳定性,可用于未来空间医学即时检测.
English Summary:
To reveal the potential myocardial and muscle injury of astronauts during long-duration spaceflight, an aptamer-based colloidal gold chromatography Strips were studied to for rapid Creatine Kinase-MB detection. Firstly, the obtained Creatine Kinase-MB aptamer was screened based on a SELEX method. And then a series of aptamer with strong affinity for Creatine Kinase-MB were selected preferably to be applied to the colloidal gold chromatography strips. By optimizing conditions such as the size of colloidal gold particle and the concentration of buffer, a method of aptamer-based lateral flow strips for rapid CK-MB detection was successfully established. The results show that, the aptamers with affinity for Creatine Kinase-MB screened by a magnetic bead-based nucleic acid aptamer screening method can be used to construct an aptamer-based colloidal gold chromatography strips. It has been successfully applied to the rapid and visual detection of Creatine Kinase-MB above 82 nM. At the same time, chromatography strips can be used for future space medical instant detection as its better specificity and stability.
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