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Development of a high-throughput and sensitive assay of fusion genes in lung cancer by array-based MALDI-TOFMS

机译：基于阵列的MALDI-TOFMS开发高通量且灵敏的肺癌融合基因检测方法

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ALK (anaplastic lymphoma kinase gene), ROS1 ( ros proto-oncogene 1 ) and RET ( ret proto-oncogene ) fusions are oncogenic drivers in non-small cell lung cancer (NSCLC). Methods like fluorescence in situ 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 ALK , ROS1 and RET fusion detection. This method was established with three fusion gene positive cell lines (H2228, ALK positive; HCC78, ROS1 positive; LC-2/AD, RET 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 ALK fusions, 1 case showed ROS1 fusions, no case showed RET fusions and 4 cases were both ALK and ROS1 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 ALK , ROS1 and RET 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.
机译：在非小细胞肺癌（NSCLC）中，ALK（间变性淋巴瘤激酶基因），ROS1（ros原癌基因1）和RET（ret原癌基因）融合物是致癌的驱动因素。荧光原位杂交（FISH）和免疫组织化学（IHC）等方法是高度敏感的方法，但其分析较为主观，劳动强度大，价格昂贵且不适用于多重融合基因筛选。这项研究旨在为ALK，ROS1和RET融合建立一种高通量，灵敏且具有成本效益的筛选方法（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）检测。用三种融合基因阳性细胞系（H2228，ALK阳性; HCC78，ROS1阳性; LC-2 / AD，RET阳性）和阴性样品建立了该方法。然后，通过Sanger测序，下一代测序（NGS）和基于阵列的MALDI-TOFMS选择并检测了34个临床样品。对结果进行比较和分析，并以Sanger测序为标准。 ALK融合7例，ROS1融合1例，RET融合无例，ALK和ROS1融合4例。结果表明，基于阵列的MALDI-TOFMS与Sanger测序和NGS 82.3％的一致性为100％。在这项研究中，我们报道了基于阵列的MALDI-TOFMS在FFPE和新鲜组织标本常规肺活检中评估ALK，ROS1和RET融合的实用性。此外，该方法还可用于疾病的诊断，监测和预后。

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