Rozkoš Tomáš, Ryška Aleš, Nová Markéta, Hornychová Helena, Krbal Lukáš, Matěj Radoslav, Laco Jan
Cesk Patol. 2017 Spring;53(2):89-96.
The aim of the retrospective part of the study was a) to select an optimal clone of immunohistochemical (IHC) antibody against the ALK protein with specificity and sensitivity high enough to use this antibody as a screening method for selecting non-small cell lung cancer (NSCLC) cases for fluorescence in situ hybridization (FISH) testing of ALK gene rearrangement and b) to determine the diagnostic yield of "small" biopsies i.e. endobronchial, transbronchial and transthoracic biopsies and cytoblocks for ALK gene rearrangement testing. The best IHC method of ALK protein detection (clone D5F3, dilution 1:100, Cell Signaling Technology, Danvers, MA, USA) was then verified in prospective routine testing of patients with NSCLC. ALK status was correlated with tumor morphology and clinical data. In the retrospective part of the study, 170 EGFR-nonmutated cases of NSCLC were IHC and FISH tested. In the prospective part, 557 cases of NSCLC were tested by IHC and 76 by FISH. There were 8/154 (5.2%) cases with ALK gene rearrangement detected in the retrospective part and 24/557(4.3 %) in the prospective part. Sensitivity and specificity of the best IHC method were 100 % and 99 % in the retrospective part and 100 % and 80 % in the prospective part. The diagnostic yield of "small" biopsies was between 74 - 80 % retrospectively, depending on IHC variant, and 88 % prospectively. No case with ALK gene rearrangement detected prospectively had EGFR mutation. A high diagnostic yield confirms that ALK status testing can be used in this type of specimen. A prevalence of 5.2 % in the retrospective part (EGFR-nonmutated cases) and 4.3 % in the prospective part (without known EGFR mutation status), tumor morphology (solid and acinar type, mucinous type or at least partial mucin production (extra- and/or intracellular) as well as lower average age and male/female ratio of patients with ALK positive tumors in the prospective part (57.5 y vs. 65.2 y, 8 men and 16 women vs. 336 men and 197 women) are consistent with global data.
a）选择一种针对ALK蛋白的免疫组化（IHC）抗体的最佳克隆，其特异性和敏感性要足够高，以便将该抗体用作筛选非小细胞肺癌（NSCLC）病例进行ALK基因重排荧光原位杂交（FISH）检测的方法；b）确定“小”活检标本，即支气管内、经支气管和经胸活检标本以及细胞块用于ALK基因重排检测的诊断率。然后，在NSCLC患者的前瞻性常规检测中验证了检测ALK蛋白的最佳免疫组化方法（克隆D5F3，稀释度1:100，美国马萨诸塞州丹弗斯的Cell Signaling Technology公司）。ALK状态与肿瘤形态和临床数据相关。在研究的回顾性部分，对170例EGFR未突变的NSCLC病例进行了免疫组化和FISH检测。在前瞻性部分，对557例NSCLC病例进行了免疫组化检测，76例进行了FISH检测。回顾性部分检测到8/154（5.2%）例有ALK基因重排，前瞻性部分为24/557（4.3%）。回顾性部分最佳免疫组化方法的敏感性和特异性分别为100%和99%，前瞻性部分为100%和80%。回顾性分析中，“小”活检标本的诊断率在74%-80%之间，取决于免疫组化变体，前瞻性分析中为88%。前瞻性检测中未发现有ALK基因重排的病例存在EGFR突变。高诊断率证实ALK状态检测可用于此类标本。回顾性部分（EGFR未突变病例）的患病率为5.2%，前瞻性部分（未知EGFR突变状态）为4.3%，肿瘤形态（实性和腺泡型、黏液型或至少部分黏液产生（细胞外和/或细胞内））以及前瞻性部分ALK阳性肿瘤患者的平均年龄较低和男女比例（57.5岁对65.2岁，8名男性和16名女性对336名男性和197名女性）与全球数据一致。
