Wu Han-Tao, Li Kun, Wang Gang, Yang Xue-Xi, Zhu Anna, Xu Xu-Ping, Li Ming, Wu Ying-Song, Liu Tian-Cai
Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University Guangzhou 510515 Guangdong P. R. China
Guangzhou Darui Biotechnology Co. Ltd. Guangzhou 510665 China.
RSC Adv. 2018 Aug 6;8(49):27935-27945. doi: 10.1039/c8ra05165h. eCollection 2018 Aug 2.
ALK (anaplastic lymphoma kinase gene), () and () fusions are oncogenic drivers in non-small cell lung cancer (NSCLC). Methods like fluorescence hybridization (FISH) and immunohistochemistry (IHC) are highly sensitive but subjectively analyzed, labor intensive, expensive and unsuitable for multiple fusion gene screening. This study aimed to establish a high-throughput, sensitive and cost-effective screening method (array-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, array-based MALDI-TOFMS) for , and fusion detection. This method was established with three fusion gene positive cell lines (H2228, positive; HCC78, positive; LC-2/AD, positive) and negative samples. Then, 34 clinical samples were selected and detected by Sanger sequencing, next generation sequencing (NGS) and array-based MALDI-TOFMS. The results were compared and analyzed and Sanger sequencing was considered the standard. 7 cases showed fusions, 1 case showed fusions, no case showed fusions and 4 cases were both and fusions. Results showed that array-based MALDI-TOFMS was 100% concordant with Sanger sequencing and NGS 82.3%. In this study, we reported the utility of array-based MALDI-TOFMS in the assessment of , and fusions in routine lung biopsies of FFPE and fresh tissue specimens. Besides, this method may also be applied to the diagnosis, monitoring and prognosis of illness.
间变性淋巴瘤激酶基因（ALK）、（此处原文缺失部分内容）和（此处原文缺失部分内容）融合是非小细胞肺癌（NSCLC）中的致癌驱动因素。荧光原位杂交（FISH）和免疫组织化学（IHC）等方法高度敏感，但分析主观、劳动强度大、成本高且不适用于多种融合基因筛查。本研究旨在建立一种高通量、灵敏且经济高效的筛查方法（基于阵列的基质辅助激光解吸电离飞行时间质谱，基于阵列的MALDI-TOFMS）用于（此处原文缺失部分内容）、（此处原文缺失部分内容）和（此处原文缺失部分内容）融合检测。该方法通过三种融合基因阳性细胞系（H2228，（此处原文缺失部分内容）阳性；HCC78，（此处原文缺失部分内容）阳性；LC-2/AD，（此处原文缺失部分内容）阳性）和阴性样本建立。然后，选择34例临床样本，通过桑格测序、二代测序（NGS）和基于阵列的MALDI-TOFMS进行检测。将结果进行比较和分析，并将桑格测序视为标准。7例显示（此处原文缺失部分内容）融合，1例显示（此处原文缺失部分内容）融合，无病例显示（此处原文缺失部分内容）融合，4例同时存在（此处原文缺失部分内容）和（此处原文缺失部分内容）融合。结果显示，基于阵列的MALDI-TOFMS与桑格测序的一致性为100%，与NGS的一致性为82.3%。在本研究中，我们报道了基于阵列的MALDI-TOFMS在福尔马林固定石蜡包埋（FFPE）和新鲜组织标本的常规肺活检中评估（此处原文缺失部分内容）、（此处原文缺失部分内容）和（此处原文缺失部分内容）融合的效用。此外，该方法还可应用于疾病的诊断、监测和预后评估。
