摘要
采用优化的碳化二亚胺法制备睾酮(TES)人工抗原,免疫新西兰大白兔制备多克隆抗体,建立间接竞争ELISA标准曲线。标准曲线在PBS中的线性检测范围为0.096～92.200 ng/mL,相关系数R2=0.972 2,半数抑制浓度(IC50)为2.800 ng/mL,最低检测限(LOD)为0.035 ng/mL;抗体与去甲睾酮的交叉反应率为22.3%,与其他检测化合物的交叉反应很小或忽略不计。羊尿20倍稀释后进行添加回收试验,其添加值与检测值的相关系数为0.985 2。因此,该免疫学方法可用于动物尿液中睾酮残留水平的定量分析。
A standard curve of indirect competitive ELISA （icELISA） was developed for the determination of testosterone （TES） residue in urine. The artificial antigen of TES was synthesized by carbodiimide method （EDC） and then used to im-mune New Zealand white rabbits for the preparation of polyclonal antibody. The results showed that the linear detecting range of the standard curve in PBS was from 0.096 to 92.200 ng/mL, the correlation coefficient （R2） was 0.972 2, the limit of de-tection （LOD） and IC50 value was 0.035 ng/mL and 2.800 ng/mL, respectively. The cross-reactivity of the antibody to nortestosterone was 22.3% and slight or negligible to other compounds. Under 20-fold dilution in sheep urine, a correlation coefficient of 0.985 2 between concentration added and concentration determined was observed. It indicated that this assay could be incorporated into a quantitative monitoring program for the rapid screening of TES residue in urine.
出处
《湖北农业科学》
北大核心
2012年第16期3554-3556,3563,共4页
Hubei Agricultural Sciences
基金
"十一五"国家科技支撑计划项目(2006BAK02A21/1)
河南省教育厅科技攻关项目(2011A230003)
关键词
睾酮
人工抗原
多克隆抗体
间接竞争ELISA
testosterone
artificial antigen
polyclonal antibody
indirect competitive ELISA
