氯霉素磁分离酶联免疫(MEIA)检测方法的建立  

Methodological study on magnetic affinity immunoassay for detecting chloramphenicol

作　　者：张波[1,2] 刘振世[1] 杨海[1] 杨祥良[1] 

机构地区：[1]华中科技大学生命科学与技术学院,湖北武汉430074 [2]北京倍爱康生物技术股份有限公司,北京100070

出　　处：《细胞与分子免疫学杂志》2011年第5期585-586,589,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：武汉市科技攻关计划项目(201020722309)

摘　　要：目的：采用磁分离酶联免疫分析法（MEIA）建立定量检测氯霉素（CAP）的一种高灵敏度方法。方法：采用免疫竞争法,用异硫氰酸荧光素（FITC）标记抗CAP抗体,碱性磷酸酶（AP）标记CAP,以羊抗FITC抗体包被的免疫磁珠作为固相分离载体,单磷酸酚酞做显色底物,建立检测CAP的MEIA法。结果：成功建立了检测CAP的MEIA法,检测时间为40 min,检测灵敏度为0.03μg/L,线性范围为0.1~8.1μg/L,批内变异系数（CV）3.9%~5.3%,批间CV 4.8%~8.1%,回收率为97%~101.5%,与国标方法液相色谱-串联质谱法的检测结果对比,相关系数r=0.9817。结论：检测CAP的MEIA法的灵敏度及准确率高、检测时间短,为食品中氯霉素残留的监测提供了一种新的免疫分析方法。AIM： To establish a sensitive immunoassay for detection chloramphenicol using magnetic beads as solid phase.METHODS： This assay employs the competitive inhibition,the FITC conjugated with anti-chloramphenicol monoclonal antibodies and the alkaline phosphatase conjugated with chloramphenicol respectively.Magnetic beads were coupled with sheep anti-FITC antibodies as solid phase.And phenolphthalein monophosphate was used as substrate to set up MEIA for detection chloramphenicol.RESULTS： MEIA for detection chloramphenicol was Established,the reaction time is 40 minutes,the sensitivity is 0.03 μg/L,the liner range is 0.1-8.1 μg/L,the intra and inter coefficient variation（CV） was 3.9%-5.3% and 4.8%-8.1% respectively and the recovery is 97%-101.5%.Comparing with national standard method of liquid chromatography-mass spectrometry,the Correlation coefficient of test results is 0.9817（r=0.9817）.CONCLUSION： The chloramphenicol MEIA method is sensitive,accurate,and fast,it provides a new method of immunoassay for the monitoring of chloramphenicol residues in food.

关 键 词：氯霉素 磁分离酶联免疫 免疫检测 

分 类 号：R392.33[医药卫生—免疫学]