摘要
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。
Objectlve To establish reaction pattern for determining three tumor markers in human sernm 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 and precision of the assay were evaluated. Correlation analysis was made between the measurements obtained by liquid protein chip and by chemiluminescence immunoassay （CLIA） or time-resolved fluorescence immunoassay （TRFIA）. Results The linear range for the measurement of CEA, AFP and tPSA were 0. 063 ~ 200, 0. 052 - 66.6, 0. 008 ~ 10 ng/ml respectively. The lower detection limit was 31.3, 13.0, 5.2 pg/ ml respectively. The intra- and inter-assay coefficients of variation were 6.2% ~ 9. 1 % and 7.5% ~ 13.5% respectively. For the detection of CEA, AFP, and tPSA in clinical serum samples, the assay had a sensitivity of 96.2% - 98.2% , specificity of 97.7% ~ 98.2%, and accuracy of 97% ~ 98%. Significant positive correlations were found between the meas- urements obtained by the liquid protein chip and by CLIA or TRFIA , all P 〈 0.00 1. Conclusions For multiple detection of tumor markers, the liquid protein chip has the advantages of wide linear range, high sensitivity, good reproducibility, simple operation and small sample amount, and would have potential in future clinical practice.
出处
《山东医药》
CAS
北大核心
2010年第47期3-6,共4页
Shandong Medical Journal
基金
中国博士后科学基金资助项目(2005037607)
关键词
液相蛋白芯片
肿瘤标志物
方法学评价
liquid protein chip
tumor markers
evaluation of methodology
