Background Liquid biopsy testing, especially molecular tumor profiling of circulating tumor DNA (ctDNA) in cell-free plasma, has received increasing interest in recent years as it serves as a reliable alternative for the detection of tumor-specific aberrations to guide treatment decision-making in oncology. Many (commercially available) applications have been developed, however, broad divergences in (pre)analytical work flows and lack of universally applied guidelines impede routine clinical implementation. In this review, critical factors in the blood-based ctDNA liquid biopsy work flow are evaluated. Content In the preanalytical phase, several aspects (e.g., blood collection tubes [BCTs], plasma processing, and extraction method) affect the quantity and quality of the circulating cell-free DNA (ccfDNA) applicable for subsequent molecular analyses and should meet certain standards to be applied in diagnostic work flows. Analytical considerations, such as analytical input and choice of assay, might vary based on the clinical application (i.e., screening, primary diagnosis, minimal residual disease [MRD], response monitoring, and resistance identification). In addition to practical procedures, variant interpretation and reporting ctDNA results should be harmonized. Collaborative efforts in (inter)national consortia and societies are essential for the establishment of standard operating procedures (SOPs) in attempts to standardize the plasma-based ctDNA analysis work flow. Summary Development of universally applicable guidelines regarding the critical factors in liquid biopsy testing are necessary to pave the way to clinical implementation for routine diagnostics.

中文翻译：

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基于循环肿瘤 DNA 的分子谱分析分析工作流程中的关键因素

背景 液体活检检测，尤其是无细胞血浆中循环肿瘤 DNA (ctDNA) 的分子肿瘤分析，近年来受到越来越多的关注，因为它是检测肿瘤特异性畸变以指导治疗决策的可靠替代方案。在肿瘤学中。许多（商业上可用的）应用程序已经开发出来，然而，（预）分析工作流程的巨大差异以及缺乏普遍适用的指南阻碍了常规临床实施。在这篇综述中，评估了基于血液的 ctDNA 液体活检工作流程中的关键因素。内容在分析前阶段，几个方面（例如，采血管[BCT]、血浆处理和提取方法）影响适用于后续分子分析的循环游离DNA（ccfDNA）的数量和质量，应满足一定的标准应用于诊断工作流程。分析考虑因素，例如分析输入和检测方法的选择，可能会根据临床应用（即筛查、初步诊断、微小残留病[MRD]、反应监测和耐药性鉴定）而有所不同。除了实际程序外，变异解释和 ctDNA 结果报告也应协调一致。国家（国际）财团和学会的合作对于建立标准操作程序 (SOP) 至关重要，以标准化基于血浆的 ctDNA 分析工作流程。摘要 制定关于液体活检关键因素的普遍适用的指南对于常规诊断的临床实施铺平道路是必要的。