The Establishment of an Enzyme linked Immunosorbent Assay for Prostate specific Antigen
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摘要: 一株 PSA单抗用于固相包被, 另一株单抗与辣根过氧化物酶偶联制备抗 PSA酶标记物, 以四甲基联苯胺 (TMB)为底物, 采用二步法, 建立了 PSA酶联免疫分析试剂盒 (EL ISA)制备方法。本方法的灵敏度为 0.2 μg/ L,批内变异系数 <5.3%, 批间变异系数 <6.4 %, 回收率为 96.6 %～10 5.5%, 特异性鉴定显示与其他肿瘤标志物无明显交叉反应。正常参考值 <4.0 μg/ L。本法所制试剂盒与 PSA- EL ISA试剂盒 (Maxim Biotech,Inc.)对照, 两者相关方程为 y=0.993x- 0.16, 相关系数 r=0.932 (n=4 8)。该方法具有简便、快速和准确的特点, 适于临床检测和科研应用。
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关键词:
- 前列腺特异性抗原(PSA) /
- 酶联免疫分析法(ELISA) /
- 前列腺癌(PCa)
Abstract: The two step assay is based on enzyme linked immunosorbent assay (ELISA). Using two anti PSA monoclonal antibodies, one of them is coated on the microtiter plate, the other is labeled with horseradish peroxidase(HRPO). TMB H 2O 2 solution is used as the substrate of HRPO. The sensitivity of the assay is 0.2 μg/L. The intra assay CVs and the inter assay CVs of 3 samples are lower than 5.3% and 6.4% respectively. The analytical recoveries are ranged from 96.6% to 105.5%. The reference cut off value from 100 healthy men is 4.0 μg/L. The correlation coefficient comparing with PSA ELISA (Maxim Biotech, Inc.) is 0.932 ( n =48). This assay is a rapid, sensitive and precise method. It is suitable for clinical and research application. -
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