液相蛋白芯片检测三种肿瘤标志物反应模式的建立及方法学评价

目的建立用液相蛋白芯片检测人血清三种肿瘤标志物的反应模式，并对其进行方法学评价。方法将癌胚抗原（CEA）、甲胎蛋白（AFP）和总前列腺特异性抗原（tPSA）的包被抗体分别交联于不同荧光编码的微球上，并自行制备生物素标记抗体和抗原标准品，建立双抗体夹心反应模式，评价液相蛋白芯片的检测范围、最低检测限、精密度等指标；将检测结果与化学发光免疫分析（CLIA）、时间分辨荧光免疫分析（TRFIA）进行相关性分析。结果CEA、AFP、tPSA的线性范围分别为0．063～200、0．052—66．6、0．008～10ng／ml；最低检测限分别为31．3、13．0、5．2pg／ml；批内变异系数（cv）为6．2％～9．1％，批间CV为7．5％-13．5％；检测100（109）份临床血清标本的灵敏度为96．2％～98．2％，特异度为97．7％～98．2％，准确度为97％-98％；液相蛋白芯片与CLIA及TRFIA结果均呈明显正相关，P均〈0．001。结论液相蛋白芯片检测多种肿瘤标志物具有线性范围广、灵敏度高、重复性好、操作简便、标本用量少等优点，具有临床应用潜力。

Establishment and evaluation of reaction pattern for the determination of three tumor markers by liquid protein chip

Objective To establish reaction pattern for determining three tumor markers in human serum by liquid protein chip and evaluate its assay performance.Methods The coating antibodies of carcinoembryonic antigen(CEA),alpha fetoprotein(AFP) and total prostate specific antigen(tPSA) were conjugated to the different fluorescent encoding microspheres.Biotinylated detection antibodies and antigen standards were prepared.A double antibody sandwich immunoassay was established.The detection range,lower detection limit ...