Introduction: Genomic variants that lead to MET proto‐oncogenem receptor tyrosine kinase (MET) exon 14 skipping represent a potential targetable molecular abnormality in NSCLC. Consequently, reliable molecular diagnostic approaches that detect these variants are vital for patient care. Methods: We screened tumor samples from patients with NSCLC for MET exon 14 skipping by using two distinct approaches: a DNA‐based next‐generation sequencing assay that uses an amplicon‐mediated target enrichment and an RNA‐based next‐generation sequencing assay that uses anchored multiplex polymerase chain reaction for target enrichment. Results: The DNA‐based approach detected MET exon 14 skipping variants in 11 of 856 NSCLC samples (1.3%). The RNA‐based approach detected MET exon 14 skipping in 17 of 404 samples (4.2%), which was a statistically significant increase compared with the DNA‐based assay. Among 286 samples tested by both assays, RNA‐based testing detected 10 positives, six of which were not detected by the DNA‐based assay. Examination of primer binding sites in the DNA‐based assay in comparison with published MET exon 14 skipping variants revealed genomic deletion involving primer binding sequences as the likely cause of false negatives. Two samples positive via the DNA‐based approach were uninformative via the RNA‐based approach due to poor‐quality RNA. Conclusions: By circumventing an inherent limitation of DNA‐based amplicon‐mediated testing, RNA‐based analysis detected a higher proportion of MET exon 14 skipping cases. However, RNA‐based analysis was highly reliant on RNA quality, which can be suboptimal in some clinical samples.

中文翻译：

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简要报告：肺癌中 MET 外显子 14 跳跃事件的基于 DNA 与基于 RNA 的检测

简介：导致 MET 原癌基因受体酪氨酸激酶 (MET) 外显子 14 跳跃的基因组变异代表了 NSCLC 中潜在的可靶向分子异常。因此，检测这些变异的可靠分子诊断方法对于患者护理至关重要。方法：我们通过使用两种不同的方法筛选来自 NSCLC 患者的肿瘤样本的 MET 外显子 14 跳跃：使用扩增子介导的靶标富集的基于 DNA 的下一代测序测定和使用的基于 RNA 的下一代测序测定。用于靶标富集的锚定多重聚合酶链反应。结果：基于 DNA 的方法在 856 份 NSCLC 样本中的 11 份（1.3%）中检测到 MET 外显子 14 跳跃变异。基于 RNA 的方法在 404 个样本中的 17 个（4.2%）中检测到 MET 外显子 14 跳跃，与基于 DNA 的检测相比，这是一个统计学上显着的增加。在通过两种测定法检测的 286 个样本中，基于 RNA 的检测检测到 10 个阳性，其中 6 个未通过基于 DNA 的检测法检测到。与已发表的 MET 外显子 14 跳跃变体相比，基于 DNA 的测定中引物结合位点的检查揭示了涉及引物结合序列的基因组缺失是假阴性的可能原因。由于 RNA 质量差，两个通过基于 DNA 的方法呈阳性的样本无法通过基于 RNA 的方法提供信息。结论：通过规避基于 DNA 的扩增子介导的测试的固有限制，基于 RNA 的分析检测到更高比例的 MET 外显子 14 跳跃病例。然而，基于 RNA 的分析高度依赖于 RNA 质量，这在某些临床样本中可能不是最理想的。