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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
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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.
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 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)
关键词 氯霉素 磁分离酶联免疫 免疫检测 chloramphenicol magnetic enzyme-linked immunoassay(MEIA) immunoassay
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