Applied Proteomics, Inc., 3545 John Hopkins Court, San Diego, CA, 92121, USA.
Applied Proteomics, Inc., 3545 John Hopkins Court, San Diego, CA, 92121, USA.
J Pharm Biomed Anal. 2018 May 30;154:85-94. doi: 10.1016/j.jpba.2018.02.038. Epub 2018 Feb 23.
Early detection of colorectal cancer (CRC) is key to reducing associated mortality. Despite the importance of early detection, approximately 40% of individuals in the United States between the ages of 50-75 have never been screened for CRC. The low compliance with colonoscopy and fecal-based screening may be addressed with a non-invasive alternative such as a blood-based test. We describe here the analytical validation of a multiplexed blood-based assay that measures the plasma concentrations of 15 proteins to assess advanced adenoma (AA) and CRC risk in symptomatic patients. The test was developed on an electrochemiluminescent immunoassay platform employing four multi-marker panels, to be implemented in the clinic as a laboratory developed test (LDT). Under the Clinical Laboratory Improvement Amendments (CLIA) and College of American Pathologists (CAP) regulations, a United States-based clinical laboratory utilizing an LDT must establish performance characteristics relating to analytical validity prior to releasing patient test results. This report describes a series of studies demonstrating the precision, accuracy, analytical sensitivity, and analytical specificity for each of the 15 assays, as required by CLIA/CAP. In addition, the report describes studies characterizing each of the assays' dynamic range, parallelism, tolerance to common interfering substances, spike recovery, and stability to sample freeze-thaw cycles. Upon completion of the analytical characterization, a clinical accuracy study was performed to evaluate concordance of AA and CRC classifier model calls using the analytical method intended for use in the clinic. Of 434 symptomatic patient samples tested, the percent agreement with original CRC and AA calls was 87% and 92% respectively. All studies followed CLSI guidelines and met the regulatory requirements for implementation of a new LDT. The results provide the analytical evidence to support the implementation of the novel multi-marker test as a clinical test for evaluating CRC and AA risk in symptomatic individuals.
早期发现结直肠癌（CRC）是降低相关死亡率的关键。尽管早期检测非常重要，但在美国，年龄在 50-75 岁之间的人群中，大约有 40%从未接受过 CRC 筛查。由于结肠镜检查和基于粪便的筛查的依从性较低，因此可以使用非侵入性替代方法，如基于血液的检测。我们在此描述了一种基于血液的多重检测的分析验证，该检测通过测量血浆中 15 种蛋白质的浓度来评估有症状患者的高级腺瘤（AA）和 CRC 风险。该检测是在电化学发光免疫分析平台上开发的，采用了四个多标志物面板，将作为实验室开发的检测（LDT）在临床上实施。根据临床实验室改进修正案（CLIA）和美国病理学家学会（CAP）的规定，使用 LDT 的美国临床实验室必须在发布患者检测结果之前，建立与分析有效性相关的性能特征。本报告介绍了一系列研究，这些研究证明了每个 15 项检测的精密度、准确性、分析灵敏度和分析特异性，这是 CLIA/CAP 要求的。此外，该报告还描述了每个检测的动态范围、平行性、对常见干扰物质的耐受性、加标回收率以及对样本冻融循环的稳定性等特征。在完成分析特性描述后，进行了一项临床准确性研究，以评估使用旨在用于临床的分析方法对 AA 和 CRC 分类器模型调用的一致性。在测试的 434 个有症状患者样本中，与原始 CRC 和 AA 检测结果的一致性分别为 87%和 92%。所有研究均遵循 CLSI 指南，并满足实施新 LDT 的监管要求。这些结果提供了分析证据，支持将新型多标志物检测作为评估有症状个体 CRC 和 AA 风险的临床检测方法。
