心脏疾病标志物cTnT和H-FABP联合检测微流体测试卡的开发及评价  

Development and evaluation of microfluidic test card for simultaneous detection of cardiac disease markers cTnT and H-FABP

作　　者：孙雪梅[1] 许强[1] 孙晓萌[1] 王轶鹏[1] 吕衍民 孙晓艳[1] 孟凡达 SUN Xuemei;XU Qiang;SUN Xiaomeng;WANG Yipeng;LYU Yanmin;SUN Xiaoyan;MENG Fanda(Institute of Basic Medicine,Shandong First Medical University/Shandong Academy of Medical Sciences,Jinan,Shandong 250062,China)

机构地区：[1]山东省医学科学院基础医学研究所/山东第一医科大学,山东济南250062

出　　处：《国际检验医学杂志》2021年第1期59-63,共5页International Journal of Laboratory Medicine

基　　金：山东省自然科学基金项目(ZR2019PH114);山东省医药卫生科技发展计划项目(2017WS075、2015WS0196)。

摘　　要：目的建立一种在自驱动微流体测试卡上快速联合检测心脏疾病标志物心肌肌钙蛋白(cTn)T和心型脂肪酸结合蛋白(H-FABP)的方法。方法使用激光切割的方式快速简便构建微流体结构,使用时间分辨微球作为标记,优化捕获抗体,标记抗体的浓度,评价测试卡的稳定性、特异性,并进行血清检测。结果该检测方法能够在10 min内完成2种心脏疾病标志物的联合检测,cTnT在0.1~100.0 ng/mL范围内有良好的线性关系,检出限为0.02 ng/mL(s/n=3);H-FABP在0.5~100.0 ng/mL范围内有良好的线性关系,检出限为0.05 ng/mL(s/n=3)。进行血清检测时,该方法与市场化检测方法比较,相关系数>0.95,相关性良好。结论该研究建立的检测方法有特异性良好、检测时间短、检测结果准确等优点,实现了心脏病标志物的联合检测,能够满足临床的需求,有良好的应用前景。Objective To establish a method for rapid detection of cardiac disease markers cardiac troponin(cTn)T and heart fatty acid binding protein(H-FABP)on self driving microfluidic test card.Methods The microfluidic structure was constructed fastly and simply by laser cutting.The time-resolved fluorescence microspheres were used as signal probes.The capture and detection antibody concentration were optimized.The stability and specificity of the test card were evaluated,and clinical serum samples were detected.Results The simultaneous detection of two cardiac disease markers could be completed within 10 min.cTnT detection had a good linear relationship in the range of 0.1-100.0 ng/mL,and the detection limit was 0.02 ng/mL(s/n=3);the linear range of H-FABP detection was 0.5-100.0 ng/mL,and the detection limit was 0.05 ng/mL(s/n=3).Compared with commercial method,the test card showed good correlation over 0.95 in clinical blood samples detection.Conclusion The detection method established in this study has the advantages of high sensitivity,short detection time and accurate detection results.It realizes the simultaneous detection of heart disease markers,which can meet the clinical needs and has a good application prospect.

关 键 词：微流体 早期诊断 联合检测 

分 类 号：R-331[医药卫生]