摘要
目的采用时间分辨荧光技术建立高灵敏的盐酸克伦特罗（CBL）间接竞争免疫分析法（CBL-TRFIA）。方法以CBL-BSA为免疫原免疫新西兰大白兔制备抗CBL抗体；CBL与卵清蛋白的联结物（CBL-OVA）包被96孔板为固相抗原，与游离CBL共同竞争有限的抗CBL抗体；用稀土离子Eu^3＋标记的羊抗兔抗体进行示踪。结果该方法的灵敏度为0．01μg／L，测量范围为0．01～25μg／L，平均回收率为99．7％，RSD3．9％，与盐酸异丙肾上腺素的交叉反应率为0．01％，与沙丁胺醇、盐酸肾上腺素、重酒石酸去甲肾上腺素无交叉反应。8条不同时间进行的CBL-TRFIA的效应点值ED80、ED50、ED20分别为（0．07±0．01）／Lg／L、（1．47±0．11）μg／L和（23．6±0．56）μg/L。尿样经TRnA和EUSA试剂盒同时检测CBL，两者的相关系数为0．932，结果相符。结论CBL-TRFIA分析方法稳定性好，可测范围宽，具有很好的应用前景。
Objective In order to provide a rapid and selectivity method for the determination of clenbuterel（ CBL）, an indirect competitive time-resolved fluoroimmunoassay （TRFIA） was developed. Methods Anti-CBL antibody, was raised by immunization against CBL-BSA in rabbits. CBL-OVA was coated by physical adsorption onto the micretitre plate, CBL or sample with CBL as a competitor. Both them were incubated with limited anti- CBL antibody, and a goat antirabbit IgG-Eu^3＋ conjugate was used as a tracer. Results The sensitivity of CBL-TRFIA was 0.01μg/L, and the recovery rate was 99.7%. RSD of CBL-TRFIA was 3.9% .The sensitivity of CBL-TRFIA provided a linear reaponse from 0.01 -25μg/L, with ED50 of （1.47± 0.11 ）μg/L or EDso of （0.07 ±0.01 ）μg/L and ED2o of （23.6 ±0.56）μg/L. The cross reactivity of the CBL-TRFIA with salbutamol, epinephrine hydrochloride and epinephrine bitartrate was negligible, while that with isoprenaline hydrochloride was 0.01%. Both CBL-TRFIA and CBL-ELISA test were applied for the quantitative measurement of CBL in the same urine, and the coefficient of correlation was 0.932. Conclusion The CBL-TRFIA could be applied to detect the CBL in urine and it is useful to screening easily for CBL contamination in meat or foods.
出处
《卫生研究》
CAS
CSCD
北大核心
2006年第6期798-801,共4页
Journal of Hygiene Research
基金
无锡市2005重大科技计划项目(No.DL050001)
