【摘要】 【目的】采用液相芯片技术同时定量测定人血清中癌胚抗原(CEA)、甲胎蛋白(AFP)和乙肝表面抗原(HBsAg),并对该方法进行评价。【方法】制备抗体交联微球及生物素标记抗体,用双抗体夹心法对临床血清标本进行测定。【结果】同时检测CEA、AFP和HBsAg时的线性范围分别为0.032～200ng/mL、0.022～55U/mL、0.26～1000ng/mL,最低检测限分别为8.90pg/mL、0.013U/mL、0.24ng/mL,批内变异系数(CV)<9%,批间CV<14%。检测CEA、AFP、HBsAg的灵敏度分别为96.4%、95.8%、92%,特异度分别为95.6%、94.2%、100%,准确度分别为96%、95%、96%。其检测结果与化学发光免疫分析法(CLIA)或时间分辨荧光免疫分析法(TRFIA)之间呈显著的等级相关关系。该方法仅需1!L标本,3h即可完成检测。【结论】液相芯片技术具有可联合检测多项指标、高通量、线性范围广、灵敏度高、重复性好、节省样品和时间等优点,具有临床应用潜力。更多还原

【Abstract】 [Objective] To simultaneously quantify carcinoembryonic antigen (CEA), alpha fetoprotein (AFP), and hepatitis B surface antigen (HBsAg) in human serum using multiplexed Luminex assay and evaluate the assay performance. [Methods] Biotinylated detection antibodies and microspheres coupled with capture antibodies were prepared. A sandwich immunoassay was established to measure the three analytes in serum samples. [Results] Linear ranges were 0.032～200 ng/mL, 0.022～55 U/mL, 0.26～1 000 ng/mL, and detection limits were 8.90 pg/mL, 0.013 U/mL, 0.24 ng/mL for CEA, AFP, and HBsAg, respectively. The intra-and inter-assay coefficients of variation were less than 9% and 14% respectively. For CEA, AFP, and HBsAg, sensitivity were 96.4%, 95.8% and 92%, specificity were 95.6%, 94.2% and 100%, and accuracy were 96%, 95% and 96%, respectively. Significant Spearman rank correlations were found between the results obtained by Luminex assay and by chemiluminescence immunoassay (CLIA) or time-resolved fluorescence immunoassay (TRFIA). Only 1 microliter of serum and 3 hours were needed for Luminex assay. [Conclusion] With the advantages of unique capability of multiplexing and high-throughput, good reproducibility, increased detection range and sensitivity, Luminex assay would have a great potential in clinical application.更多还原