热休克蛋白90α荧光免疫层析检测方法的建立

摘要

目的:建立人血清中热休克蛋白90α(HSP90α)的荧光免疫层析定量检测方法.方法:采用荧光免疫层析技术,对荧光微球制备、最佳反应时间、线性范围、精密性、回收率、临床检测等方面进行系统研究.结果:确定了各反应条件,最佳反应时间为5 min.线性范围0.39~100 ng/ml.精密性质控品高值(30 ng/ml)CV=8.14%;低值(10 ng/ml)CV=9.26%,高、中、低值血清的回收率为100.56%、99.76%、94.1%;与国外试剂盒(ELISA检测方法)平行检测40份临床血清,结果显示两者相关系数为0.968 5.结论:初步建立的方法具有良好的精密性和线性,临床符合率能满足临床检测技术要求,有望完善后报批用于临床.%Objective:To establish quantitative a fluorescence immunochromatographic assay detection method for the heat shock protein 90α(HSP90α)in human serum.Methods: Indirect the technology of fluorescence microshers immunochromatographic assay was used to research fluorescent microsphere activation,ensure optimal testing time,determine linearity range,precision,recovery experiments,testing clinical samples and that sort of thing in the fluorescence immunochromatographicassay experiment.Results: In all the reaction conditions,it was determined that the optimal teaction time was 5 minutes,and in the best line range (0.39-100 ng/ml) precision quality control materials of high recovery (30 ng/ml) Intra-assay CV was 8.14%;quality control materials of low recovery (10 ng/ml) Intra-assay CV was 9.26%.With high,medium and low recovery of serum recovery was 100.56%,99.76%,94.1%,separately.Foreign kit(ELISA) for detection of 40 cancer patients clinical serum,the result showed that the correlation was 0.968 5.Conclusion: Establishing the method initially quantitative fluorescence immunochromatographic assay for the heat shock protein 90α(HSP90α)in human serum have high performance and linear,clinic accordance rate also can satisfy the demand of clinic quantitative detection,and will improve hopeful to applied to clinic after apprroved.