快速检测心型脂肪酸结合蛋白的荧光免疫微流体测试卡

引用本文: 孟凡达, 霍卫松, 贺美琳, 李昊, 廉洁, 石西增, 高云华. 快速检测心型脂肪酸结合蛋白的荧光免疫微流体测试卡[J]. 分析化学, 2016, 44(4): 633-639. doi: 10.11895/j.issn.0253-3820.160045 shu

Citation: MENG Fan-Da, HUO Wei-Song, HE Mei-Lin, LI Hao, LIAN Jie, SHI Xi-Zeng, GAO Yun-Hua. A Microfluidic Fluorescence Immunoassay Test Card for Rapid Detection of Heart-type Fatty Acid Binding Protein[J]. Chinese Journal of Analytical Chemistry, 2016, 44(4): 633-639. doi: 10.11895/j.issn.0253-3820.160045 shu

快速检测心型脂肪酸结合蛋白的荧光免疫微流体测试卡

孟凡达 ^1,2, ,
霍卫松 ^1, ,
贺美琳 ^1,2, ,
李昊 ^1,2, ,
廉洁 ^3, ,
石西增 ^4, ,
高云华 ^{1,
,}

1.
¹ 中国科学院理化技术研究所光化学转换和功能材料重点实验室, 北京 100190;
2.
² 中国科学院大学, 北京 100049;
3.
³ 中国人民公安大学刑事科学技术系, 北京 100038;
4.
⁴ 东莞博识生物科技有限公司, 东莞 523808

通讯作者: 高云华

基金项目:
本文系东莞市重大科技项目(No.2014215102) (No.2014215102)

SEM技术创新基金(No.14C26214402622)资助 (No.14C26214402622)

摘要: 研制出一种以时间分辨荧光微球作为标记,自驱动快速检测H-FABP的荧光免疫微流体测试卡。利用激光切割法在双面胶上简便、快速地切割出所设计的微通道结构,并采用激光切割法制作出聚甲基丙烯酸甲酯(PMMA)测试卡底板及上盖。使用提拉涂膜的方法在PMMA底板表面修饰马来酸酐官能团,有效地解决了捕获抗体在PMMA表面的固定问题。使用等离子体处理测试卡上盖改善其亲水性,使液体能够在微通道内自行流动。使用此测试卡可以实现对心型脂肪酸结合蛋白(H-FABP)的快速检测,线性检测范围为0.5~100 ng/mL,检出限为0.1 ng/mL(S/N=3),检测时间少于10 min,批内相对标准偏差(RSD)<10%,批间RSD<15%。本方法具有灵敏度高、检测时间短、结果准确等优点,可以满足临床检测的需求,具有良好的应用前景。

关键词:

English

A Microfluidic Fluorescence Immunoassay Test Card for Rapid Detection of Heart-type Fatty Acid Binding Protein

1.
¹ Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190, China;
2.
² University of Chinese Academy of Sciences, Beijing 100049, China;
3.
³ Department of Forensic Science, People's Public Security University of China, Beijing 100038, China;
4.
⁴ Dongguan Bosh Biotechnologies, LTD, Dongguan 523808, China

Corresponding author: 高云华

Received Date: 16 January 2016
Fund Project:
本文系东莞市重大科技项目(No.2014215102)

SEM技术创新基金(No.14C26214402622)资助

Abstract: A kind of easily assembled self-driving test card was developed for the rapid detection and quantification of heart-type fatty acid binding protein(H-FABP) by using time-resolved fluorescence microspheres as signal probes. It was easy to make microfluidic channel structures with double-sided adhesive and cut out of test card substrate and cover based on polymethyl methacrylate(PMMA) material by laser cutting technology. The membrane of polystyrene containing maleic anhydride functional groups was coated on PMMA plate surface by dip-coating method. Further, the capture antibody was immobilized effectively on PMMA surface by covalent binding with maleic anhydride groups. The cover was treated with plasma treatment to improve the hydrophilic ability, so that liquid could flow smoothly in the microchannel. The whole analysis process could be completed within 10 min. There was a very good linear correlation between response and H-FABP concentration in the range of 0.5-100 ng/mL(R²=0.9966), and the detection limit was 0.1 ng/mL(S/N=3). The intra-assay precision(RSD) was less than 10% and the inter-assay RSD was less than 15%. This detection method has the advantages such as high sensitivity, fast detection and good accuracy.

Key words:

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

快速检测心型脂肪酸结合蛋白的荧光免疫微流体测试卡

通讯作者: 高云华

English

A Microfluidic Fluorescence Immunoassay Test Card for Rapid Detection of Heart-type Fatty Acid Binding Protein

Corresponding author: 高云华

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

快速检测心型脂肪酸结合蛋白的荧光免疫微流体测试卡

通讯作者: 高云华

English

A Microfluidic Fluorescence Immunoassay Test Card for Rapid Detection of Heart-type Fatty Acid Binding Protein

Corresponding author: 高云华

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs