下一代测序（NGS）允许快速分析多个基因以检测临床可行的变异体。本研究报告了定向泛癌症NGS检测面板CANSeqTMKids的分子分析确认，以便针对儿童恶性肿瘤进行分子分析。分析确认包括从去身份化的临床标本中提取的DNA和RNA，包括福尔马林固定石蜡包埋（FFPE）组织、骨髓和全血，以及商用参考材料。面板的DNA组分可以评估130个基因，以检测与儿童恶性肿瘤相关的单核苷酸变异（SNVs）、插入和缺失（INDELs）和91个基因的融合变异。在使用低至20%肿瘤组织含量和5 ng核酸输入的条件下进行了优化。数据的评估确定了大于99%的准确性、灵敏度、重复性和再现性。检测限定为SNVs和INDELs的5%等位基因分数，基因扩增的5个副本以及基因融合的1,100个读数。通过自动化文库制备来提高筛查效率。总之，CANSeqTMKids可以从不同样本来源对儿童恶性肿瘤进行全面的分子分析，具有高质量和快速的反应时间。版权所有©2023 Schilter、Smith、Nie、Stoll、Felix、Jarzembowski和Reddi。

Next-Generation Sequencing (NGS) allows rapid analysis of multiple genes for the detection of clinically actionable variants. This study reports the analytical validation of a targeted pan cancer NGS panel CANSeqTMKids for molecular profiling of childhood malignancies. Analytical validation included DNA and RNA extracted from de-identified clinical specimens including formalin fixed paraffin embedded (FFPE) tissue, bone marrow and whole blood as well as commercially available reference materials. The DNA component of the panel evaluates 130 genes for the detection of single nucleotide variants (SNVs), Insertion and Deletions (INDELs), and 91 genes for fusion variants associated with childhood malignancies. Conditions were optimized to use as low as 20% neoplastic content with 5 ng of nucleic acid input. Evaluation of the data determined greater than 99% accuracy, sensitivity, repeatability, and reproducibility. The limit of detection was established to be 5% allele fraction for SNVs and INDELs, 5 copies for gene amplifications and 1,100 reads for gene fusions. Assay efficiency was improved by automation of library preparation. In conclusion, the CANSeqTMKids allows for the comprehensive molecular profiling of childhood malignancies from different specimen sources with high quality and fast turnaround time.Copyright © 2023 Schilter, Smith, Nie, Stoll, Felix, Jarzembowski and Reddi.