The multi-target stool DNA (mt-sDNA) test screens for colorectal cancer (CRC) by analyzing DNA methylation/mutation and hemoglobin markers to algorithmically derive a qualitative result. A new panel of highly discriminant candidate methylated DNA markers (MDMs) was recently developed. Performance of the novel MDM panel, with hemoglobin, was evaluated in a simulated screening population using archived stool samples weighted to early-stage CRC and prospectively collected advanced precancerous lesions (APLs). Marker selection (MSS) and separate preliminary independent verification studies (VS) were conducted utilizing samples from multi-center, case-control studies. Sample processing included targeted MDM capture, bisulfite conversion, and MDM quantitation. Fecal hemoglobin was quantified using enzyme-linked immunosorbent assay. Samples were stratified into 75%/25% training-testing sets; model outcomes were cross-validated 1,000 times. All laboratory operators were blinded. The MSS included 232 cases (120 CRC/112 APLs) and 490 controls. The VS featured 210 cases (112 CRC/98 APLs) and 567 controls; APLs were 86.7% adenomas and 13.3% sessile serrated lesions (SSLs). Average age was 65.5 (cases) and 63.2 (controls) years. Mean sensitivity in the VS from cross-validation was 95.2% for CRC and 57.2% for APLs, with specificities of 89.8% (no CRC/APLs) and 92.4% (no neoplasia). Subgroup analyses showed CRC sensitivities of 93.4% (Stage I) and 94.2% (Stage II). APL sensitivity was 82.9% for high-grade dysplasia, 73.4% for villous lesions, 49.8% for tubular lesions, and 30.2% for SSLs. These data support high sensitivity and specificity for a next-generation mt-sDNA test panel. Further evaluation of assay performance will be characterized in a prospective, multicenter clinical validation study (NCT04144738).

中文翻译：

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新一代多靶点粪便DNA面板准确检测结直肠癌和晚期癌前病变

多目标粪便 DNA (mt-sDNA) 测试通过分析 DNA 甲基化/突变和血红蛋白标记物，通过算法得出定性结果，从而筛查结直肠癌 (CRC)。最近开发了一组新的高度辨别性候选甲基化 DNA 标记 (MDM)。使用加权早期 CRC 的存档粪便样本和前瞻性收集的晚期癌前病变 (APL)，在模拟筛查人群中评估新型 MDM 组合（含血红蛋白）的性能。利用多中心病例对照研究的样本进行了标记选择（MSS）和单独的初步独立验证研究（VS）。样品处理包括目标 MDM 捕获、亚硫酸氢盐转化和 MDM 定量。使用酶联免疫吸附测定法对粪便血红蛋白进行定量。样本被分层为 75%/25% 训练测试集；模型结果交叉验证了 1,000 次。所有实验室操作员均处于失明状态。MSS 包括 232 例病例（120 例 CRC/112 例 APL）和 490 例对照。VS 包含 210 个病例（112 个 CRC/98 APL）和 567 个对照；APL 中 86.7% 为腺瘤，13.3% 为无蒂锯齿状病变 (SSL)。平均年龄为 65.5（病例）和 63.2（对照）岁。交叉验证的 VS 中 CRC 的平均敏感性为 95.2%，APL 的平均敏感性为 57.2%，特异性为 89.8%（无 CRC/APL）和 92.4%（无肿瘤）。亚组分析显示 CRC 敏感性为 93.4%（第一阶段）和 94.2%（第二阶段）。APL 对高度不典型增生的敏感性为 82.9%，对绒毛状病变的敏感性为 73.4%，对管状病变的敏感性为 49.8%，对 SSL 的敏感性为 30.2%。这些数据支持下一代 mt-sDNA 测试小组的高灵敏度和特异性。对检测性能的进一步评估将在一项前瞻性、多中心临床验证研究 (NCT04144738) 中进行表征。