Detection of immunoglobulin E using an aptamer based dot-blot assay  

作　　者：WANG YiXian YE ZunZhong YING YiBin 

机构地区：[1]College of Biosystems Engineering and Food Science, Zhejiang University

出　　处：《Chinese Science Bulletin》2013年第24期2938-2943,共6页

基　　金：supported by the National Natural Science Foundation of China (30825027);Special Fund for Agro-Scientific Research in the Public Interest (200903009)

摘　　要：A novel aptamer based dot-blot assay for the detection of immunoglobulin E (IgE) was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose membrane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3',5,5'-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol/L to 1 μmol/L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol/L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.A novel aptamer based dot-blot assay for the detection of immunoglobulin E （IgE） was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose mem- brane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3＇,5,5＇-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol/L to 1 μmol/L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol/L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.

关 键 词：免疫球蛋白E 实验检测 基础 适体 辣根过氧化物酶 硝酸纤维素膜 生物素标记 印迹 

分 类 号：R392[医药卫生—免疫学]