ALK、ROS1 和 RET 重排分别发生在 5%、2% 和 1% 的非小细胞肺癌 (NSCLC) 中。通过免疫组织化学 (IHC) 检测 ALK 和 ROS1 融合蛋白已得到验证，可用于快速患者筛查，但 ROS1 融合需要通过其他技术进行确认，并且没有 RET IHC 测试可用于临床。我们在此报告 HTG EdgeSeq 检测的有用性，一种无需提取 RNA 的测试，将定量核酸酶保护测定与 NGS 相结合，用于检测“现实生活”小 NSCLC 样本中的 ALK、ROS1 和 RET 融合体。从 11 个中心总共收集了 203 个 FFPE 样本。其中包括 143 个重排 NSCLC（87 个 ALK、39 个 ROS1、17 个 RET）和 60 个 ALK-ROS1-RET 阴性对照。该检测的特异性为 98%，对 ALK、ROS1 和 RET 融合的敏感性分别为 80%、94% 和分别为100%。在 19 个 HTG 检测假阴性样本中，分析前条件被确定为影响检测效率的主要因素。总体而言，HTG EdgeSeq 检测提供了与其他 RNA 测序技术相当的灵敏度和特异性，其优点是可用于非常小的和旧的样本是多中心收集的。

ALK, ROS1 and RET rearrangements occur respectively in 5%, 2% and 1% non-small cell lung cancers (NSCLC). ALK and ROS1 fusion proteins detection by immunohistochemistry (IHC) has been validated for rapid patient screening, but ROS1 fusions need to be confirmed by another technique and no RET IHC test is available for clinical use.We report herein the usefulness of the HTG EdgeSeq Assay, an RNA extraction-free test combining a quantitative nuclease protection assay with NGS, for the detection of ALK, ROS1 and RET fusions from 'real-life' small NSCLC samples. A total of 203 FFPE samples were collected from 11 centers. They included 143 rearranged NSCLC (87 ALK, 39 ROS1, 17 RET) and 60 ALK-ROS1-RET negative controls.The assay had a specificity of 98% and a sensitivity for ALK, ROS1 and RET fusions of 80%, 94% and 100% respectively. Among the 19 HTG-assay false negative samples, the preanalytical conditions were identified as the major factors impacting the assay efficiency.Overall, the HTG EdgeSeq assay offers comparable sensitivities and specificity than other RNA sequencing techniques, with the advantage that it can be used on very small and old samples collected multicentrically.