Next-Generation Sequencing of Liquid-Based Cytology Non-Small Cell Lung Cancer Samples

被引:49
|
作者
Reynolds, Jordan P. [1 ]
Zhou, Yaolin [1 ]
Jakubowski, Maureen A. [1 ]
Wang, Zhen [1 ]
Brainard, Jennifer A. [1 ]
Klein, Roger D. [1 ]
Farver, Carol F. [1 ]
Almeida, Francisco A. [2 ]
Cheng, Yu-Wei [1 ]
机构
[1] Cleveland Clin, Robert J Tomsich Pathol & Lab Med Inst, Cleveland, OH 44106 USA
[2] Cleveland Clin, Resp Inst, Cleveland, OH 44106 USA
关键词
epidermal growth factor receptor; fine-needle aspiration biopsy; Ion Torrent; MiSeq; next-generation sequencing; non-small cell lung cancer; ENDOBRONCHIAL ULTRASOUND; EGFR MUTATIONS; SPECIMENS; CARCINOMA; ASPIRATE; COLLEGE; SLIDES; FNA; DNA;
D O I
10.1002/cncy.21812
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
BACKGROUND: The detection of mutated epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC) with residual cell pellets derived from liquid-based cytology (LBC) samples (eg, endoscopic ultrasound-guided fine-needle aspiration) has been validated with allele-specific polymerase chain reaction. The aim of this study was to validate next-generation sequencing (NGS) technology for detecting gene mutations with residual cell pellets from LBC. METHODS: Archived DNA extracted from LBC samples of adenocarcinoma stored in PreservCyt with a known EGFR mutation status was retrieved. Genomic DNA was multiplex-amplified and enriched with Ion AmpliSeq Cancer Hotspot Panel v2 chemistry and the OneTouch 2 instrument; this was followed by semiconductor sequencing on the Ion Personal Genome Machine platform. The mutation hotspots of 6 NSCLC-related genes (BRAF, EGFR, ERBB2, KRAS, MET, and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit a [PIK3CA]) were analyzed with NextGENe and Torrent Suite bioinformatics tools. RESULTS: The commonly identified EGFR sequence changes, including 4 L858R mutations, 3 exon 19 deletions, and 1 exon 20 insertion, were in 100% concordance between the assay platforms. Less common NSCLC variants were also found in the mutation hotspots of ERBB2, KRAS, MET, and PIK3CA genes. CONCLUSIONS: NSCLC mutation analysis using NGS can be successfully performed on residual cell pellets derived from LBC samples. This approach allows the simultaneous examination of multiple mutation hotspots in a timely manner to improve patient care. (C) 2017 American Cancer Society.
引用
收藏
页码:178 / 187
页数:10
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