荧光免疫层析定量检测髓过氧化物酶方法的建立

Establishment of quantitative detection of myeloperoxidase by fluorescence immunochromatography

目的:通过荧光免疫层析技术初步建立MPO荧光免疫层析定量检测方法。方法:采用羧基荧光微球,利用EDC一步法偶联标记抗体,对过程中EDC量、标记蛋白量与标记时间、保护液进行研究,选出最佳条件,并对线性范围、灵敏度、精密性等进行性能评价。结果:确定偶联过程中EDC 160μg,标记蛋白125μg、标记1. 5 h;线性范围3. 125~600 ng/ml,最低检出限0. 9 ng/ml;精密性质控品(高、中、低)批内、批间变异系数均小于10%;通过与国外试剂盒(ELISA法)检测比对45份临床血清,相关性良好,R^2=0. 979 5。结论:成功建立荧光免疫层析快速定量检测MPO的方法,为冠心病的辅助诊断提供一种技术手段。

Objective:To establish quantitative detection method of MPO by using fluorescence immunochromato-graphy.Methods:The carboxyl fluorescent microspheres were coupled with labelled antibodies by one-step EDC method.In order to investigate the most stable reaction conditions,the EDC quantity,labeled protein quantity,labeling time and complex solution were studied.Then to evaluate the linear range,sensitivity and precision.Results:The coupling process of EDC amount was 160μg,the amount of labeled protein was 125μg and marker time 1.5 h.The linear range was 3.125~600 ng/ml,the lowest detection concentration was 0.9 ng/ml.Both variation coefficient in intra and inter batches of precision controlled products that divided from high,medium and low concentration were less than 10%.By detected 45 clinical serums with comparing foreign kit(ELISA),it showed good correlation,which R 2 could reach 0.979 5.Conclusion:We established the rapid quantitative detection method of MPO by fluorescence immunochromatography,which provides a reference diagnosis for coronary heart disease.