From the Institute for Experimental Pathology, ARUP Laboratories, Salt Lake City, Utah (Wilcock, Moore, Rowe, Mahlow, Jedrzkiewicz, Cleary, Lomo, Ruano, Gering, Bradshaw, Maughan, Tran, Burlingame, Davis, Affolter, Albertson, Adelhardt, Kim, Coleman, Deftereos, Gulbahce, Sirohi).
The Department of Pathology, University of Utah and ARUP Laboratories, Salt Lake City (Mahlow, Jedrzkiewicz, Cleary, Lomo, Ruano, Affolter, Albertson, Adelhardt, Kim, Coleman, Deftereos, Gulbahce, Sirohi).
Arch Pathol Lab Med. 2023 Dec 1;147(12):1402-1412. doi: 10.5858/arpa.2022-0372-OA.
CONTEXT.—: Quantitative imaging is a promising tool that is gaining wide use across several areas of pathology. Although there has been increasing adoption of morphologic and immunohistochemical analysis, the adoption of evaluation of fluorescence in situ hybridization (FISH) on formalin-fixed, paraffin-embedded tissue has been limited because of complexity and lack of practice guidelines.
OBJECTIVE.—: To perform human epidermal growth factor receptor 2 (HER2) FISH validation in breast carcinoma in accordance with the American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) 2018 guideline.
DESIGN.—: Clinical validation of HER2 FISH was performed using the US Food and Drug Administration-approved dual-probe HER2 IQFISH (Dako, Carpinteria, California) with digital scanning performed on a PathFusion (Applied Spectral Imaging, Carlsbad, California) system. Validation parameters evaluated included z-stacking, classifier, accuracy, precision, software, and hardware settings. Finally, we evaluated the performance of digital enumeration on clinical samples in a real-world setting.
RESULTS.—: The accuracy samples showed a final concordance of 95.3% to 100% across HER2 groups 1 to 5. During clinical implementation for HER2 groups 2, 3, and 4, we achieved a final concordance of 76% (95 of 125). Of these cases, only 8% (10 of 125) had discordances with clinical impact that could be identified algorithmically and triaged for manual review.
CONCLUSIONS.—: Digital FISH enumeration is a useful tool to improve the efficacy of HER2 FISH enumeration and capture genetic heterogeneity across HER2 signals. Excluding cases with high background or poor image quality and manual review of cases with ASCO/CAP group discordances can further improve the efficiency of digital HER2 FISH enumeration.
定量成像技术是一种很有前途的工具，在病理学的多个领域得到了广泛的应用。尽管形态学和免疫组织化学分析的应用越来越多，但由于其复杂性和缺乏实践指南，福尔马林固定、石蜡包埋组织中荧光原位杂交（FISH）的评估应用受到限制。
根据美国临床肿瘤学会/美国病理学家协会（ASCO/CAP）2018 年指南，对乳腺癌中人类表皮生长因子受体 2（HER2）FISH 进行验证。
HER2 FISH 的临床验证使用美国食品和药物管理局批准的双探针 HER2 IQFISH（Dako，加利福尼亚州卡平特里亚）进行，数字扫描在 PathFusion（Applied Spectral Imaging，加利福尼亚州卡尔斯巴德）系统上进行。评估的验证参数包括 z 堆叠、分类器、准确性、精密度、软件和硬件设置。最后，我们在真实环境中评估了数字计数在临床样本中的性能。
在 HER2 1 至 5 组中，准确性样本的最终一致性为 95.3%至 100%。在 HER2 2、3 和 4 组的临床实施过程中，我们达到了 76%的最终一致性（125 例中有 95 例）。在这些病例中，只有 8%（125 例中有 10 例）的病例具有可以通过算法识别并进行人工审查的临床影响不一致。
数字 FISH 计数是提高 HER2 FISH 计数效率和捕获 HER2 信号遗传异质性的有用工具。排除背景高或图像质量差的病例，以及对 ASCO/CAP 组不一致的病例进行人工审查，可以进一步提高数字 HER2 FISH 计数的效率。
