组织了用于pan-TRK免疫组织化学（IHC）染色的比利时环试验，以协调pan-TRKIHC染色方案和解释。作为一种参考方法，选择BenchmarkUltra平台上的VENTANA泛TRK测定(克隆EPR17341)。选择了六个样本：2个阴性，2个融合阳性和2个具有野生型内源性TRK表达的样品。每个参与实验室使用其常规pan-TRKIHC对载玻片进行染色并报告其结果。此外,他们被要求从每个病例中返回一张TRK染色的载玻片.协调实验室评估了这些幻灯片，将它们与参考方法进行比较并评分。在环试验期间使用两个克隆:A7H6R(细胞信号转导)和EPR17341(Abcam/Ventana)。七个协议达到了足够的性能标记，建议三个实验室进一步优化方案。在具有生理性TRK表达的情况下，pan-TRKIHC的解释被证明具有挑战性。此外,取决于NTRK融合伙伴，染色可以在强度和染色模式两者中强烈变化。使用基于EPR17341克隆的Ventana即用型系统并使用推荐的协议设置的实验室得分最高。然而,给定一些小的优化，所有实验室在技术染色和后续评估方面得分都很高。
A Belgian ring
trial for pan-TRK immunohistochemistry (IHC) staining was organised to harmonise pan-TRK IHC staining protocols and interpretation. As a reference method, the VENTANA pan-TRK Assay (clone EPR17341) on the Benchmark Ultra platform was selected. Six samples were selected: 2 negative, 2 fusion positive and 2 samples with wild-type endogenous TRK expression. Each participating laboratory stained the slides using their routine pan-TRK IHC and reported their results. In addition, they were asked to return one TRK-stained slide from each case. The coordinating lab evaluated these slides, compared them with the reference method and scored them. Two clones were used during the ring
trial: A7H6R (Cell Signaling) and EPR17341 (Abcam/Ventana). Seven protocols achieved a sufficient performance mark, and three labs were advised to further optimise the protocol. Interpretation of pan-TRK IHC proved to be challenging in cases with physiological TRK expression. In addition, depending on the NTRK fusion partner, the staining can vary strongly in both intensity and staining pattern. Labs using the Ventana ready-to-use system based on the EPR17341 clone and using the recommended protocol settings scored best. However, given some small optimisation, all labs scored well on the technical staining and the succeeding evaluation.
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