基于免疫磁球的埃博拉病毒糖蛋白高灵敏比色检测  

Colorimetric Detection of Ebola Glycoprotein Based on Immunomagnetic Microspheres

作　　者：熊俊逸 胡姣 夏骊 庞代文[1] 张志凌[1] 

机构地区：[1]生物医学分析化学教育部重点实验室武汉大学化学与分子科学学院,湖北武汉430072

出　　处：《分析科学学报》2017年第5期626-630,共5页Journal of Analytical Science

基　　金：国家自然科学基金(No.21475099)

摘　　要：用免疫磁球捕获埃博拉病毒糖蛋白,与生物素化抗体形成免疫夹心复合物,然后链霉亲和素标记辣根过氧化物酶(SA-HRP)催化3,3',5,5'-四甲基联苯胺(TMB)进行显色,通过370 nm处吸光度对病毒糖蛋白进行定量。对免疫磁球进行了免疫荧光表征,通过对照实验验证方法的可靠性,并对检测条件进行了优化。结果表明,吸光度与病毒糖蛋白浓度在1.0~25.0 ng/m L内呈线性关系,检出限达到0.18 ng/m L。该方法重现性较好,特异性好,抗干扰能力强,可实现复杂样品中埃博拉病毒的检测。Ebola glycoprotein was captured by immunomagnetic microspheres, followed by formation of an immune sandwich complex with biotinylated antibody. Then, 3,3＇, 5,5＇-tetramethylbenzidine （TMB） was developed by streptavidin-horseradish peroxidase （SA-HRP） to form a colored product,which had a strong absorption at 370 nm. Finally,Ebola glycoprotein was quantified with the absorbance at 370 nm. Immunomagnetic microspheres was characterized by immune fluorescence. Reliability of detection principle was proved by control experiments. At the optimized conditions,the absorbance was linear with the concentration of Ebola glycoprotein in the range of 1. 0-25. 0 ng/mL, with a detection limit of 0.18 ng/mL. This method showed good reproducibility, high specificity and nice capacity of resisting disturbance.

关 键 词：埃博拉病毒 磁球 比色检测 

分 类 号：O657.3[理学—分析化学]