快速检测心型脂肪酸结合蛋白的荧光免疫微流体测试卡

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

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。

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 was0. 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.