评估循环肿瘤DNA（ctDNA）测序在精准肿瘤学中的分析有效性，这一成果由美国食品药品监督管理局Joshua Xu、澳大利亚新南威尔士大学Timothy R. Mercer和美国阿肯色大学医学院Donald J. Johann Jr研究组合作取得。2021年4月12日出版的《自然-生物技术》杂志发表了这一最新研究成果。

研究人员对五个行业领先ctDNA分析进行多站点、跨平台评估。研究通过模拟、合成DNA掺入实验对标准化、来自细胞系的参考样品进行了验证，并对ctDNA测序工作流程的每个阶段进行了评估。在高于0.5％的变异等位基因频率时，所有五种检测方法均以高灵敏度、精确度和可重复性检测到ctDNA突变；而低于此限制时，检测变得不可靠且检测之间差异很大，尤其是在输入样本有限的情况下。

遗漏突变（假阴性）比错误突变（假阳性）更常见，这表明有效采集稀有ctDNA片段是ctDNA分析的主要难题。对ctDNA分析的分析性能进行综合评估可为最佳实践指南提供信息，并为精确肿瘤学提供资源。

研究人员表示，循环肿瘤DNA测序已应用于精准肿瘤学，但对ctDNA测定的准确性、灵敏性和可重复性了解甚少。

附：英文原文

Title: Evaluating the analytical validity of circulating tumor DNA sequencing assays for precision oncology

Author: Ira W. Deveson, Binsheng Gong, Kevin Lai, Jennifer S. LoCoco, Todd A. Richmond, Jeoffrey Schageman, Zhihong Zhang, Natalia Novoradovskaya, James C. Willey, Wendell Jones, Rebecca Kusko, Guangchun Chen, Bindu Swapna Madala, James Blackburn, Igor Stevanovski, Ambica Bhandari, Devin Close, Jeffrey Conroy, Michael Hubank, Narasimha Marella, Piotr A. Mieczkowski, Fujun Qiu, Robert Sebra, Daniel Stetson, Lihyun Sun, Philippe Szankasi, Haowen Tan, Lin-ya Tang, Hanane Arib, Hunter Best, Blake Burgher, Pierre R. Bushel, Fergal Casey, Simon Cawley, Chia-Jung Chang, Jonathan Choi, Jorge Dinis, Daniel Duncan, Agda Karina Eterovic, Liang Feng, Abhisek Ghosal, Kristina Giorda, Sean Glenn, Scott Happe, Nathan Haseley, Kyle Horvath, Li-Yuan Hung, Mirna Jarosz, Garima Kushwaha, Dan Li, Quan-Zhen Li, Zhiguang Li, Liang-Chun Liu, Zhichao Liu, Charles Ma, Christopher E. Mason, Dalila B. Megherbi, Tom Morrison, Carlos Pabn-Pea, Mehdi Pirooznia, Paula Z. Proszek, Amelia Raymond, Paul Rindler, Rebecca Ringler, Andreas Scherer, Rita Shaknovich, Tieliu Shi, Melissa Smith, Ping Song, Maya Strahl, Venkat J. Thodima, Nikola Tom, Suman Verma

Issue&Volume: 2021-04-12

Abstract: Circulating tumor DNA (ctDNA) sequencing is being rapidly adopted in precision oncology, but the accuracy, sensitivity and reproducibility of ctDNA assays is poorly understood. Here we report the findings of a multi-site, cross-platform evaluation of the analytical performance of five industry-leading ctDNA assays. We evaluated each stage of the ctDNA sequencing workflow with simulations, synthetic DNA spike-in experiments and proficiency testing on standardized, cell-line-derived reference samples. Above 0.5% variant allele frequency, ctDNA mutations were detected with high sensitivity, precision and reproducibility by all five assays, whereas, below this limit, detection became unreliable and varied widely between assays, especially when input material was limited. Missed mutations (false negatives) were more common than erroneous candidates (false positives), indicating that the reliable sampling of rare ctDNA fragments is the key challenge for ctDNA assays. This comprehensive evaluation of the analytical performance of ctDNA assays serves to inform best practice guidelines and provides a resource for precision oncology. Reliable detection of mutations below 0.5% variant allele frequency remains a key challenge for circulating tumor DNA sequencing assays.

DOI: 10.1038/s41587-021-00857-z

Source: https://www.nature.com/articles/s41587-021-00857-z