免疫组织化学是筛选结直肠癌和甲状腺乳头状癌 BRAF V600E 突变的一种可行方法。
Immunohistochemistry is a feasible method to screen BRAF V600E mutation in colorectal and papillary thyroid carcinoma.
机构信息
Department of Pathology, The Affiliated Hospital of Qingdao University, 16 Jiangsu Road, Qingdao 266003, China.
Department of Pneumology, The Affiliated Hospital of Qingdao University, 16 Jiangsu Road, Qingdao 266003, China.
出版信息
Exp Mol Pathol. 2018 Aug;105(1):153-159. doi: 10.1016/j.yexmp.2018.07.006. Epub 2018 Jul 19.
AIMS
To investigate whether immunohistochemistry (IHC) could be used to screen BRAF mutation status in formalin-fixed paraffin-embedded (FFPE) colorectal carcinoma (CRC) and papillary thyroid carcinoma (PTC) samples.
METHODS
Eight surgical resected samples, including 2 CRCs with mutated BRAF V600E, 2 PTCs with mutated BRAF V600E, 2 CRCs with wild-type BRAF and 2 PTCs with wild-type BRAF, were selected to explore the optimized IHC conditions for BRAF V600E (VE1) immunostaining using BenchmarkXT automated immunostainer VENTANA Medical System. BRAF V600E status was tested by optimized IHC and ARMS-PCR methods in 255 samples (123 CRCs and 132 PTCs). Sanger sequencing was performed to validate the BRAF V600E status if discordant results were found between optimized IHC and ARMS-PCR methods.
RESULTS
Antigen retrieval time in 32 min and 64 min showed satisfactory intensity and homogeneity of BRAF V600E staining in CRC and PTC samples, respectively. The concordance between IHC and ARMS/Sanger sequencing was 99.2% (122/123) in CRCs and 96.2% (127/132) in PTCs. In CRCs, the sensitivity of IHC staining for BRAF V600E was 100% (3/3) and the specificity was 99.1% (119/120). In PTCs, the sensitivity was 100% (106/106) and specificity was 80.8% (21/26). The overall concordance across all cases was 97.6% (249/255).
CONCLUSION
The appropriate antigen retrieval protocol is critical for accurate analysis of BRAF V600E status by Benchmark XT automated immunostainer. IHC is a suitable method to screen BRAF V600E mutation in FFPE samples of CRCs and PTCs.
目的
研究免疫组织化学（IHC）是否可用于筛选福尔马林固定石蜡包埋（FFPE）结直肠癌（CRC）和甲状腺乳头状癌（PTC）样本中的 BRAF 突变状态。
方法
选择 8 例手术切除样本，包括 2 例 BRAF V600E 突变的 CRC、2 例 BRAF V600E 突变的 PTC、2 例野生型 BRAF 的 CRC 和 2 例野生型 BRAF 的 PTC，以探索使用 BenchmarkXT 自动免疫染色仪（VENTANA Medical System）对 BRAF V600E（VE1）免疫染色进行优化的 IHC 条件。在 255 例样本（123 例 CRC 和 132 例 PTC）中，采用优化的 IHC 和 ARMS-PCR 方法检测 BRAF V600E 状态。如果优化的 IHC 和 ARMS-PCR 方法之间存在不一致的结果，则进行 Sanger 测序以验证 BRAF V600E 状态。
结果
在 CRC 和 PTC 样本中，抗原修复时间为 32 分钟和 64 分钟时，BRAF V600E 染色的强度和均匀性均令人满意。在 CRC 中，IHC 与 ARMS/Sanger 测序的一致性为 99.2%（122/123），在 PTC 中为 96.2%（127/132）。在 CRC 中，IHC 染色检测 BRAF V600E 的灵敏度为 100%（3/3），特异性为 99.1%（119/120）。在 PTC 中，灵敏度为 100%（106/106），特异性为 80.8%（21/26）。所有病例的总体一致性为 97.6%（249/255）。
结论
对于 Benchmark XT 自动免疫染色仪的准确分析，适当的抗原修复方案至关重要。IHC 是筛选 CRC 和 PTC 中 BRAF V600E 突变的合适方法。
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