核糖核酸
液体活检
数字聚合酶链反应
RNA剪接
分子生物学
癌症研究
化学
生物
癌症
生物化学
基因
遗传学
聚合酶链反应
作者
Ana Giménez-Capitán,Estela Sánchez-Herrero,Lucía Robado de Lope,Andrés Aguilar-Hernández,Ivana Sullivan,Virginia Calvo,Irene Moya-Horno,Santiago Viteri,Carlos Cabrera,Cristina Aguado,Noelia Armiger,Joselyn Valarezo,Clara Mayo-de-las-Casas,Noemi Reguart,Martyna Filipska,Mariano Provencio,Atocha Romero,Miguel Angel Molina-Vila
标识
DOI:10.1002/1878-0261.13468
摘要
ALK, ROS1, and RET fusions and MET∆ex14 variant associate with response to targeted therapies in non-small-cell lung cancer (NSCLC). Technologies for fusion testing in tissue must be adapted to liquid biopsies, which are often the only material available. In this study, circulating-free RNA (cfRNA) and extracellular vesicle RNA (EV-RNA) were purified from liquid biopsies. Fusion and MET∆ex14 transcripts were analyzed by nCounter (Nanostring) and digital PCR (dPCR) using the QuantStudio® System (Applied Biosystems). We found that nCounter detected ALK, ROS1, RET, or MET∆ex14 aberrant transcripts in 28/40 cfRNA samples from positive patients and 0/16 of control individuals (70% sensitivity). Regarding dPCR, aberrant transcripts were detected in the cfRNA of 25/40 positive patients. Concordance between the two techniques was 58%. Inferior results were obtained when analyzing EV-RNA, where nCounter often failed due to a low amount of input RNA. Finally, results of dPCR testing in serial liquid biopsies of five patients correlated with response to targeted therapy. We conclude that nCounter can be used for multiplex detection of fusion and MET∆ex14 transcripts in liquid biopsies, showing a performance comparable with next-generation sequencing platforms. dPCR could be employed for disease follow-up in patients with a known alteration. cfRNA should be preferred over EV-RNA for these analyses.
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