摘要
建立背景荧光猝灭-免疫层析法(bFQICA)定量检测降钙素原(PCT)的方法。利用双抗体夹心法原理,将合适质量浓度的捕获抗体、检测抗体分别固定在试纸条和微孔内,层析时与样本形成抗体-抗原-抗体的夹心结构。测定PCT的质量浓度范围为0. 27~20 ng/mL,检出限为0. 27 ng/mL,加标回收率为94. 0%~94. 6%,试剂的批间和批内相对标准偏差均小于15%,与C-反应蛋白(CRP,10. 0μg/mL)、人血清淀粉样蛋白A (SAA,10. 0μg/mL)均无交叉反应,样本中常见的干扰物在所测定的质量浓度下对PCT定量检测无明显影响,用该方法与90例临床血清样本测得值进行对比,检测结果相关性良好(r=0. 9633,P <0. 01)。
A method with simple operation, high sensitivity, strong anti-interference and rapid quantitative detection of procalcitonin(PCT) was established. PCT was detected by background fluorescence quenching-immunochromatography(bFQICA). The principle of double antibody sandwich method was used to fix the appropriate concentration of capture antibody and tracer antibody in the test strip and microwell,respectively. The sandwich-antibody-antibody sandwich structure was formed with the sample upon chromatography. The concentration of PCT in this detection system ranged from 0. 27 to 20 ng/m L. The minimum detection limit was 0. 27 ng/m L,and the recovery was 94. 0%-94. 6%,the relative standard deviation of the batch and intra-assay of the three batches of reagents was less than15%,and there was no cross-reaction with C-reactive protein(CRP,10. 0 μg/m L) and human serum amyloid A(SAA,10. 0 μg/m L). The common interferents in the sample had no significant effect on the quantitative detection of PCT at the determined concentrations. This method was used to test 90 clinical serum samples. The measured results were compared with the clinical values, and the correlation was good(r = 0. 9633,P < 0. 01).
引文
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