使用 ThinPrep 细胞学制片 FISH 法检测非小细胞肺癌中的 ALK 状态。

ALK status testing in non-small-cell lung carcinoma by FISH on ThinPrep slides with cytology material.

机构信息

*Departments of Molecular Pathology, Cytopathology, and Anatomic Pathology, Robert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland, Ohio; †Departments of Translational Hematology and Oncology Research, and Solid Tumor Oncology, Taussig Cancer Institute, Cleveland, Ohio; and ‡Respiratory Institute, Cleveland Clinic Foundation, Cleveland, Ohio.

出版信息

J Thorac Oncol. 2014 Apr;9(4):464-8. doi: 10.1097/JTO.0000000000000104.

DOI:10.1097/JTO.0000000000000104

PMID:24736067

Abstract

INTRODUCTION

Oncogenic anaplastic lymphoma kinase (ALK) gene rearrangements in non-small-cell lung carcinomas (NSCLC) provide the basis for targeted therapy with crizotinib and other specific ALK inhibitors. Treatment eligibility is conventionally determined by the Food and Drug Administration-approved companion diagnostic fluorescence in situ hybridization (FISH) assay on paraffin-embedded tissue (PET). On limited samples such as fine needle aspiration-derived cytoblocks, FISH for ALK is often uninformative. FISH performed on liquid-based ThinPrep slides (ThinPrep-FISH) may represent a robust alternative.

METHODS

Two hundred thirty cytology samples from 217 patients with advanced NSCLC, including a consecutive series of 179 specimens, were used to generate matched ThinPrep slides and paraffin cytoblocks. The same ThinPrep slides used for cytologic diagnosis were assessed by standard ALK break-apart two-color probe FISH, after etching of tumor areas. Ultrasensitive ALK immunohistochemistry (IHC) on corresponding cytoblocks [D5F3 antibody, OptiView signal amplification] served as the reference data set.

RESULTS

ThinPrep-FISH ALK signals were robust in 228 of 230 cases and not compromised by nuclear truncation inherent in paraffin-embedded tissue-FISH; only two samples displayed no signals. Nine of 178 informative cases (5%) in the consecutive series and 18 of 228 informative cases (7.8%) overall were ALK rearranged by ThinPrep-FISH. In 154 informative matched ThinPrep-FISH and cytoblock-IHC samples, 152 were concordant (10, 6.5% ALK status positive; 142, 92.2% ALK status negative), and two (1.3%) were ThinPrep-FISH positive but IHC negative (sensitivity 100%, specificity 98.6%, overall agreement 98.7%).

CONCLUSION

Detection of ALK gene rearrangements in liquid cytology ThinPrep slides derived from patients with NSCLC can be confidently used for clinical ALK molecular testing.

摘要

简介

致癌性间变性淋巴瘤激酶（ALK）基因重排在非小细胞肺癌（NSCLC）中为克唑替尼和其他特定 ALK 抑制剂的靶向治疗提供了基础。治疗资格通常通过食品和药物管理局批准的石蜡包埋组织（PET）荧光原位杂交（FISH）检测来确定。在有限的样本（如细针抽吸衍生的细胞块）上，ALK 的 FISH 通常没有信息。液体基础的 ThinPrep 载玻片（ThinPrep-FISH）上的 FISH 可能是一个强大的替代方法。

方法

217 例晚期 NSCLC 患者的 230 例细胞学样本，包括连续的 179 例标本，用于生成匹配的 ThinPrep 载玻片和石蜡细胞学块。对相同的 ThinPrep 载玻片进行标准的 ALK 断裂分离双色探针 FISH 检测，在肿瘤区域蚀刻后。相应细胞块上的超敏性 ALK 免疫组织化学（IHC）[D5F3 抗体，OptiView 信号放大]作为参考数据集。

结果

230 例中有 228 例 ThinPrep-FISH ALK 信号稳定，不受石蜡包埋组织-FISH 固有的核截断的影响；只有两个样本没有信号。连续系列中的 178 例有意义的病例中有 9 例（5%），总共 228 例有意义的病例中有 18 例（7.8%）通过 ThinPrep-FISH 显示为 ALK 重排。在 154 例有意义的匹配的 ThinPrep-FISH 和细胞块-IHC 样本中，152 例结果一致（10 例，阳性率为 6.5%；142 例，阴性率为 92.2%），2 例（1.3%）为 ThinPrep-FISH 阳性但 IHC 阴性（敏感性 100%，特异性 98.6%，总一致性 98.7%）。