【摘要】 目的建立利用液相芯片技术定量检测人血清中癌胚抗原(CEA)、甲胎蛋白(AFP)和神经原特异性烯醇化酶(NSE)的反应模式,并对该方法各项指标进行评价。方法制备抗体交联微球及生物素标记抗体,用双抗夹心法检测临床血清标本。结果同时检测CEA、AFP和NSE时的线性范围分别为0.078～200ng/ml、0.025～25U/ml、0.146～75ng/ml,最低检测限分别为39.1pg/ml、0.016U/ml、0.073ng/ml,分析内精密度<10%,分析间精密度<15%。检测CEA、AFP、NSE的灵敏度分别为97.2%、100%、93.5%,特异度分别为96.4%、97.7%、97.0%,准确度分别为96.9%、98.4%、95.3%。检测结果与化学发光免疫分析法(CLIA)呈显著的等级相关关系,而且仅需1!l标本,3h就可以完成检测。结论液相芯片技术具有可联合检测多项指标、高通量、线性范围宽、灵敏度高、重复性好和节省样品和时间等优点,具有极大的临床应用潜力。更多还原

【Abstract】 Objective To measure the levels of carcinoembryonic antigen (CEA), alpha fetoprotein (AFP), and neuron specific enolase (NSE) in human serum using multiplexed bead-based immunoassay and evaluate the performance of this method. Methods Antibody-conjugated microspheres and biotinylated detection antibodies were prepared. The levels of CEA, AFP and NSE in clinical serum samples were also measured using a sandwich immunoassay. Results The linear range for the measurement of CEA, AFP and NSE was 0.078～200 ng/ml, 0.025～25 U/ml, and 0.146～75 ng/ml, respectively. The lowest detection limit for CEA, AFP, and NSE was 39.1 pg/ml, 0.016 U/ml, and 0.073 ng/ml, respectively. The coefficient of variation was less than 10% (intra-assay) and 15% (inter-assay). This method has a sensitivity of 97.2% (CEA), 100% (AFP), 93.5% (NSE), specificity of 96.4% (CEA), 97.7% (AFP), 97.0% (NSE), and an accuracy of 96.9% (CEA), 98.4% (AFP) and 95.3% (NSE). Result obtained from multiplexed bead-based immunoassay was compatible with chemiluminescence immunoassay (CLIA). However, only 1 microtiter of serum and 3 hours were required for the multiplexed bead-based immunoassay. Conclusion The multiplexed bead-based immunoassay is a sensitive, reproducible and a high-throughput method. This method may further be developed for clinical application.更多还原