Bead-based ELISA for validation of ovarian cancer early detection markers
Bead-based ELISA for validation of ovarian cancer early detection markers
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DOI：
10.1158/1078-0432.ccr-05-2007
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发表时间：
2006-04-01
影响因子：
11.5
通讯作者：
Urban, N
中科院分区：
文献类型：
--
作者：
Scholler, N;Crawford, M;Urban, N
目的：通过免疫测定法验证卵巢癌早期检测生物标志物的努力受到可用样本量有限的挑战。我们试图开发一种高效的测定血清中CA 125的方法，评估其重现性、有效性和性能，并测试其与人附睾蛋白4（HE 4）（一种有前途的新型卵巢癌标志物）多重结合的潜力。四对市售的抗-CA 125抗体和一对抗-CA 125抗体。在基于微珠的平台上评价HE 4抗体测量已知抗原浓度的准确度。两个最好的对进行了进一步评估的再现性，有效性，并能够区分从卵巢癌病例（n = 66）和妇女没有卵巢癌（n = 125）之间的盲法血清样品。两种基于CA 125微珠的测定具有高度的可重复性（重复之间的总体相关性>= 0.95;变异系数< 0.2），并且与研究标准CA 125 II RIA强相关（相关性>= 0.9）。基于受试者操作特征分析（曲线下面积，AUC，为0.85和0.84），他们区分卵巢癌病例与非病例的能力接近于CA 125 II RIA（AUC，0.87）。基于HE 4微珠的测定显示出较低的再现性，但在受试者操作特征分析中得到的AUC为0.89。结论：优化后的两种CA 125微球检测方法的检测结果与标准CA 125 Ⅱ RIA方法一致，可与HE 4微球检测方法联用，且每种检测方法仅需15 μ L样品。
Purpose: Efforts to validate ovarian cancer early detection biomarkers with immunoassays are challenged by the limited specimen volumes available. We sought to develop a specimen-efficient assay to measure CA125 in serum, assess its reproducibility, validity, and performance, and test its potential for multiplexing and combining with human epididymis protein 4 (HE4), a promising novel ovarian cancer marker.Experimental Design: Four pairs of commercially available anti-CA125 antibodies and one pair of anti-HE4 antibodies were evaluated for accuracy in measuring known concentrations of antigen on a bead-based platform. The two best pairs were further assessed for reproducibility, validity, and the ability to discriminate between blinded serum samples obtained from ovarian cancer cases (n = 66) and women without ovarian cancer (n = 125).Results: Suitability for use in a bead-based assay varied across CA125 antibody pairs. Two CA125 bead-based assays were highly reproducible (overall correlations between replicates >= 0.95; coefficients of variation < 0.2) and strongly correlated with the research standard CA125II RIA (correlations >= 0.9). Their ability to distinguish ovarian cancer cases from non-cases based on receiver operating characteristic analyses (area under the curve, AUC, of 0.85 and 0.84) was close to that of the CA125II RIA (AUC, 0.87). The HE4 bead-based assay showed lower reproducibility but yielded an AUC of 0.89 in receiver operating characteristics analysis. Multiplexing was not possible but a composite marker including CA125 and HE4 achieved an AUC of 0.91.Conclusion: Optimization procedures yielded two bead-based assays for CA125 that perform comparably to the standard CA125II RIA, which could be combined with an HE4 bead-based assay to improve diagnostic performance, and requires only 15 mu L of sample each.