下一代靶向扩增子测序（NG-TAS）：一种优化的方案和计算流程，用于经济有效地对循环肿瘤 DNA 进行分析。

Next Generation-Targeted Amplicon Sequencing (NG-TAS): an optimised protocol and computational pipeline for cost-effective profiling of circulating tumour DNA.

机构信息

Department of Oncology and Cancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Cambridge, CB2 0RE, UK.

Breast Cancer Programme, Cancer Research UK Cambridge Cancer Centre, Cambridge University Hospitals NHS Foundation Trust, Cambridge, CB2 2QQ, UK.

出版信息

Genome Med. 2019 Jan 4;11(1):1. doi: 10.1186/s13073-018-0611-9.

Circulating tumour DNA (ctDNA) detection and monitoring have enormous potential clinical utility in oncology. We describe here a fast, flexible and cost-effective method to profile multiple genes simultaneously in low input cell-free DNA (cfDNA): Next Generation-Targeted Amplicon Sequencing (NG-TAS). We designed a panel of 377 amplicons spanning 20 cancer genes and tested the NG-TAS pipeline using cell-free DNA from two HapMap lymphoblastoid cell lines. NG-TAS consistently detected mutations in cfDNA when mutation allele fraction was > 1%. We applied NG-TAS to a clinical cohort of metastatic breast cancer patients, demonstrating its potential in monitoring the disease. The computational pipeline is available at https://github.com/cclab-brca/NGTAS_pipeline .

循环肿瘤 DNA（ctDNA）检测和监测在肿瘤学中有巨大的潜在临床应用价值。我们在这里描述了一种快速、灵活且具有成本效益的方法，用于同时分析低输入游离 DNA（cfDNA）中的多个基因：下一代靶向扩增子测序（NG-TAS）。我们设计了一个包含 377 个扩增子的面板，跨越 20 个癌症基因，并使用来自两个 HapMap 淋巴母细胞系的游离 DNA 对 NG-TAS 管道进行了测试。当突变等位基因分数 > 1%时，NG-TAS 能够一致地检测到 cfDNA 中的突变。我们将 NG-TAS 应用于转移性乳腺癌患者的临床队列，证明了其在监测疾病方面的潜力。该计算管道可在 https://github.com/cclab-brca/NGTAS_pipeline 获得。