检验医学与临床2016，Vol.13Issue(7)：873-875,3.DOI:10.3969/j.issn.1672-9455.2016.07.004
双抗夹心法酶联免疫吸附试验影响因素探析
Exploration and analysis on influence factors in double antibody sandwich enzyme-linked immunosorbent assay
摘要
Abstract
Objective To explore and analyze the influence of blocking of bovine serum albumin ,tween 20 and its concentration ,sample adding mode of sample and detecting antibody and incubating temperature on the detection results of the double antibody sandwich enzyme‐linked immunosorbent assay .Methods Detecting human IL‐1Ra cy‐tokines by the double antibody sandwich enzyme‐linked immunosorbent assay was used as a model to measure the in‐fluence of the closure condition ,tween 20 and its different concentrations ,separated incubation and co‐incubation of sample and detecting antibody ,incubation temperature on the relation between standard substance concentration and OD value .Results R2 was 0 .995 7 when the enzyme‐link was not closed ,the concentration of tween 20 was 0 .05%and incubating temperature was 37℃ ;R2 by blocking BSA was 0 .981 1 ;when the concentration of tween 20 was 0 .005% or 0 .5% ,R2 was 0 .991 5 and 0 .986 8 respectively ;R2 when the sample and the detection antibody were in‐cubated together for 1 h or 2 h was 0 .964 5 and 0 .874 2 respectively ;R2 was 0 .996 1 when incubating at 25 ℃ .Con‐clusion The BSA blocking step does not improve the detection sensitivity for low concentration samples ;the sample , detection antibody and enzyme diluent must contain Tween 20 and the concentration less than 0 .05% is most appro‐priate;the co‐incubation of sample and detection antibody could affect the detection of the high concentration of sam‐ple;the enzyme‐linked detection sensitivity has no difference under the conditions of incubation temperature of 37 ℃and 25 ℃ ,but the background of OD value could be reduced in incubation at 25 ℃ .关键词
影响因素/标准操作流程/双抗夹心法酶联免疫吸附试验Key words
influence factors/standard operating procedure/double antibody sandwich ELISA引用本文复制引用
何莉,孙海,胡松,莫春芬,郭慧杰,赖翼,刘阳,邹强,罗兴燕..双抗夹心法酶联免疫吸附试验影响因素探析[J].检验医学与临床,2016,13(7):873-875,3.基金项目
国家自然科学基金资助项目（81302786；81402944）；四川省教育厅科研资助项目（15ZB0233；15ZB0235）；成都医学院自然基金资助项目（CYZ14-001）。 （）
