磁微粒化学发光酶免疫分析法检测血清f-PSA方法学建立(1)
摘要:本研究建立了一种定量分析人血清中游离前列腺特异性抗原(f-PSA)的高灵敏度磁微粒化学发光酶免疫分析方法。采用了一步双抗体夹心法,将单克隆抗体连接在磁微粒表面,以辣根过氧化物酶(Horseradish Peroxidase, HRP)为标记酶,结合鲁米诺(luminol)高灵敏化学发光反应为检测体系进行深入研究。建立检测试剂的主要指标:检测范围0.02~50ng/ml;最低检出限0.02ng/ml,分析内变异<4.7%;回收率在90%~110%之间。对510例样本进行测定临床研究,与Roche检测结果符合程度高,相关系数达到0.995。结果表明,该检测方法稳定,可靠,在临床分析人血清中f-PSA的辅助诊断前列腺癌方面具有很高的应用价值。
关键词:游离前列腺特异抗原,磁微粒化学发光酶免疫分析法,肿瘤标志物
Key words: prostate specific antigen,microbeads-based chemiluminescence immunoassay. Tumor marker
Gao wei
Kaifeng City Maternity Hospital
[Abstract]A sensitive chemiluminescence enzyme immunoassay based on magnetic micro-beads was established to detect the free prostate specific antigen (f-PSA) in human serum. One antigen was coated on the magnetic micro-beads and the other was labled with horseradish peroxidase(HRP), with the highly sensitive Luminol chemiluminescence reaction as the detection system. The linear range of this method for PSA was 0.02~50ng/ml with the limit detection of 0.02 ng. The intra-assay coefficients of variation is lower than 4.7% and the percent recovery was in the range of 90%~110%. F-PSA in 510 clinical samples were detected with this method. Compared with Roche chemiluminescence immunoassays, the correlation coefficient of 0.995 was obtained. The results demonstrated that this method is reliable and shows proposed potential application in the clinical analysis of f-PSA for prostate cancer diagnosis. ......
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