A highly specific and sensitive massive parallel sequencer-based test for somatic mutations in non-small cell lung cancer.
A highly specific and sensitive massive parallel sequencer-based test for somatic mutations in non-small cell lung cancer.
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DOI：
10.1371/journal.pone.0176525
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Hagiwara K
中科院分区：
文献类型：
--
作者：
Inoue Y;Shiihara J;Miyazawa H;Ohta H;Higo M;Nagai Y;Kobayashi K;Saijo Y;Tsuchida M;Nakayama M;Hagiwara K
非小细胞肺癌（NSCLC）的分子靶向治疗阐明了突变检测在选择治疗方案时的重要性。因此，迫切需要进行多基因突变测试。我们开发了一种基于下一代测序仪 (NGS) 的多基因测试，名为 MINtS，用于研究细胞学标本和速冻组织样本中的驱动突变。 MINtS 用于研究 DNA 中的 EGFR、KRAS、BRAF 基因以及 RNA 中的 ERBB2、ALK、ROS1 和 RET 融合基因。我们注重高特异性和敏感性（≥0.99），甚至纳入了癌细胞含量为 1% 的样本。 MIntS 能够在一次运行中测试 100 多个样品，从而可以处理提交给中心实验室的大量样品，并降低单个样品的成本。我们研究了 96 个细胞学样本和 190 个手术切除的组织，两者都是在日常临床实践中分离的。通过细胞学样本，我们比较了MINtS和PNA-LNA PCR钳夹试验的EGFR突变结果，结果一致99%。在速冻组织样本中，使用 DNA 和 RNA 成功分析了 188/190 (99%) 个样本的所有基因。然后，我们使用在临床实践中连续分离的 200 个细胞学样本来评估 RNA 质量。使用我们的程序，196 个样本 (98%) 提供了适合使用 MIntS 进行分析的高质量 RNA。我们得出的结论是，MINtS 测试系统对于使用细胞学样本和速冻组织样本分析“可成药”基因是可行的。 MIntS 将满足只能通过细胞学样本进行基因检测的患者的需求。
Molecular targeting therapy for non-small cell lung cancer (NSCLC) has clarified the importance of mutation testing when selecting treatment regimens. As a result, multiple-gene mutation tests are urgently needed. We developed a next-generation sequencer (NGS)-based, multi-gene test named the MINtS for investigating driver mutations in both cytological specimens and snap-frozen tissue samples. The MINtS was used to investigate the EGFR, KRAS, BRAF genes from DNA, and the ERBB2, and the ALK, ROS1, and RET fusion genes from RNA. We focused on high specificity and sensitivity (≥0.99) and even included samples with a cancer cell content of 1%. The MINtS enables testing of more than 100 samples in a single run, making it possible to process a large number of samples submitted to a central laboratory, and reducing the cost for a single sample. We investigated 96 cytological samples and 190 surgically resected tissues, both of which are isolated in daily clinical practice. With the cytological samples, we compared the results for the EGFR mutation between the MINtS and the PNA-LNA PCR clamp test, and their results were 99% consistent. In the snap-frozen tissue samples, 188/190 (99%) samples were successfully analyzed for all genes investigated using both DNA and RNA. Then, we used 200 cytological samples that were serially isolated in clinical practice to assess RNA quality. Using our procedure, 196 samples (98%) provided high-quality RNA suitable for analysis with the MINtS. We concluded that the MINtS test system is feasible for analyzing “druggable” genes using cytological samples and snap-frozen tissue samples. The MINtS will fill a needs for patients for whom only cytological specimens are available for genetic testing.