Scholler Nathalie, Crawford Meghan, Sato Alicia, Drescher Charles W, O'Briant Kathy C, Kiviat Nancy, Anderson Garnet L, Urban Nicole
Translational Outcomes Research Laboratory, Fred Hutchinson Cancer Research Center, Public Health Sciences, and Harborview Medical Center, University of Washington, Seattle, Washington, USA.
Clin Cancer Res. 2006 Apr 1;12(7 Pt 1):2117-24. doi: 10.1158/1078-0432.CCR-05-2007.
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.
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).
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.
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 muL of sample each.
利用免疫测定法验证卵巢癌早期检测生物标志物的工作受到可用样本量有限的挑战。我们试图开发一种样本高效的检测方法来测量血清中的CA125，评估其重现性、有效性和性能，并测试其与人类附睾蛋白4（HE4，一种有前景的新型卵巢癌标志物）进行多重检测和联合检测的潜力。
评估了四对市售抗CA125抗体和一对抗HE4抗体在基于微珠的平台上测量已知浓度抗原的准确性。进一步评估了两对最佳抗体的重现性、有效性以及区分从卵巢癌病例（n = 66）和无卵巢癌女性（n = 125）获得的盲法血清样本的能力。
各抗CA125抗体对在基于微珠的检测中的适用性各不相同。两种基于微珠的CA125检测方法具有高度重现性（重复检测之间的总体相关性≥0.95；变异系数<0.2），并且与研究标准CA125II RIA高度相关（相关性≥0.9）。根据受试者工作特征分析，它们区分卵巢癌病例与非病例的能力（曲线下面积，AUC，分别为0.85和0.84）接近CA125II RIA（AUC，0.87）。基于微珠的HE4检测方法重现性较低，但在受试者工作特征分析中AUC为0.89。无法进行多重检测，但包括CA125和HE4的复合标志物AUC达到0.91。
优化程序产生了两种基于微珠的CA125检测方法，其性能与标准CA125II RIA相当，可与基于微珠的HE4检测方法联合使用以提高诊断性能，且每种检测仅需15μL样本。
