With this external quality assessment (EQA) scheme, we aim to investigate the diagnostic performance of the currently available methods for the detection of ALK alterations in non-small cell lung cancer on a national scale, namely, in situ hybridization (ISH), immunohistochemistry (IHC), and RNA/DNA sequencing (NGS). The EQA scheme cohort consisted of ten specimens, including four ALK positive and six ALK negative samples, which were thoroughly pretested using IHC, ISH, and RNA/DNA NGS. Unstained tumor sections were provided to the 57 participants, and the results were retrieved via an online questionnaire. ISH was used by 29, IHC by 38, and RNA/DNA sequencing by 19 participants. Twenty-eight institutions (97%) passed the ring trial using ISH, 33 (87%) by using IHC, and 18 (95%) by using NGS. The highest sensitivity and interrater agreement (Fleiss ‘ kappa) was observed for RNA/DNA sequencing (99%, 0.975), followed by ISH (94%, 0.898) and IHC (92%, 0.888). However, the proportion of samples that were not evaluable due to bad tissue quality was also higher for RNA/DNA sequencing (4%) compared with ISH (0.7%) and IHC (0.5%). While all three methods produced reliable results between the different institutions, the highest sensitivity and concordance were observed for RNA/DNA sequencing. These findings encourage the broad implementation of this method in routine diagnostic, although the application might be limited by technical capacity, economical restrictions, and tissue quality of formalin-fixed samples.

通过这种外部质量评估 (EQA) 计划，我们旨在调查目前可用的方法在全国范围内检测非小细胞肺癌中 ALK 改变的诊断性能，即原位杂交 (ISH)、免疫组织化学(IHC) 和 RNA/DNA 测序 (NGS)。EQA 方案队列由 10 个样本组成，包括 4 个 ALK 阳性样本和 6 个 ALK 阴性样本，这些样本使用 IHC、ISH 和 RNA/DNA NGS 进行了彻底的预测试。向 57 名参与者提供未染色的肿瘤切片，并通过在线问卷检索结果。29 人使用 ISH，38 人使用 IHC，19 名参与者使用 RNA/DNA 测序。28家（97%）使用ISH通过环试，33家（87%）使用IHC，18家（95%）使用NGS。RNA/DNA 测序的灵敏度和交互者间一致性 (Fleiss ' kappa) 最高 (99%, 0.975)，其次是 ISH (94%, 0.898) 和 IHC (92%, 0.888)。然而，与 ISH (0.7%) 和 IHC (0.5%) 相比，RNA/DNA 测序 (4%) 由于组织质量差而无法评估的样本比例也更高。虽然所有三种方法在不同机构之间都产生了可靠的结果，但 RNA/DNA 测序的灵敏度和一致性最高。这些发现鼓励在常规诊断中广泛实施这种方法，尽管应用可能会受到技术能力、经济限制和福尔马林固定样本的组织质量的限制。与 ISH (0.7%) 和 IHC (0.5%) 相比，RNA/DNA 测序 (4%) 由于组织质量差而无法评估的样本比例也更高。虽然所有三种方法在不同机构之间都产生了可靠的结果，但 RNA/DNA 测序的灵敏度和一致性最高。这些发现鼓励在常规诊断中广泛实施这种方法，尽管应用可能会受到技术能力、经济限制和福尔马林固定样本的组织质量的限制。与 ISH (0.7%) 和 IHC (0.5%) 相比，RNA/DNA 测序 (4%) 由于组织质量差而无法评估的样本比例也更高。虽然所有三种方法在不同机构之间都产生了可靠的结果，但 RNA/DNA 测序的灵敏度和一致性最高。这些发现鼓励在常规诊断中广泛实施这种方法，尽管应用可能会受到技术能力、经济限制和福尔马林固定样本的组织质量的限制。