Identification of Factors Affecting the Success of Next-Generation Sequencing Testing in Solid Tumors
Identification of Factors Affecting the Success of Next-Generation Sequencing Testing in Solid Tumors
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DOI：
10.1093/ajcp/aqv023
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发表时间：
2016-02-01
影响因子：
3.5
通讯作者：
Chen, Hui
中科院分区：
文献类型：
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作者：
Goswami, Rashmi S.;Luthra, Rajyalakshmi;Chen, Hui
目的：临床实验室正在迅速实施下一代测序（NGS）测试进行突变分析，但有几个关于成功结果的样本质量的指导方针。方法：我们旨在建立实体瘤成功NGS的组织质量参数，并提高NGS性能。结果：使用50个基因热点突变面板对2013年检测的614例临床病例进行分析，发现NGS分析不成功的主要原因是DNA小于10 ng（91%，67/74）与极小和低细胞样本相关。高成功率与切除术（333/342，97%）和活检肿瘤大于10 mm（2）（77/77，100%）相关。NGS可以成功地在用基于甲酸的脱钙程序处理的骨标本上进行（8/11，73%）。肿瘤类型和石蜡块年龄不影响成功。我们证明了NGS可以在DNA小于10 ng的样品上进行。对2014年使用优化的工作流程测试的408例病例进行的分析显示，NGS成功率从88%提高到95%（387/408），
Objectives: Clinical laboratories are rapidly implementing next-generation sequencing (NGS) tests for mutation analysis, but there are few guidelines regarding sample quality for successful results.Methods: We aimed to establish tissue quality parameters for successful NGS in solid tumors and to improve NGS performance.Results: Analysis of 614 clinical cases tested in 2013 using a 50-gene hotspot mutation panel identified the major cause for unsuccessful NGS analysis was DNA less than 10 ng (91%, 67/74) associated with extremely small and low cellularity samples. High success rates were associated with resection procedures (333/342, 97%) and biopsied tumor larger than 10 mm(2) (77/77, 100%). NGS can be successfully performed on bone specimens processed with formic acid based decalcification procedures (8/11, 73%). Tumor type and paraffin block age did not affect success. We demonstrated that NGS can be carried out on samples with less than 10 ng DNA. Analysis of 408 cases tested in 2014 using an optimized work-flow showed improved NGS success rates from 88% to 95% (387/408) with pronounced improvement among tiny (