PMID- 26477969 OWN - NLM STAT- MEDLINE DCOM- 20161013 LR - 20220310 IS - 1872-8332 (Electronic) IS - 0169-5002 (Linking) VI - 90 IP - 3 DP - 2015 Dec TI - Comparing four different ALK antibodies with manual immunohistochemistry (IHC) to screen for ALK-rearranged non-small cell lung cancer (NSCLC). PG - 492-8 LID - S0169-5002(15)30071-4 [pii] LID - 10.1016/j.lungcan.2015.10.002 [doi] AB - OBJECTIVES: Anaplastic lymphoma kinase (ALK)-rearranged non-small cell lung cancer (NSCLC) screening is essential to its treatment such as crizotinib. Different assays have been developed to detect ALK rearrangements, such as fluorescence in situ hybridization (FISH), reverse transcriptase-PCR (RT-PCR), and immunohistochemistry (IHC). However, ALK detection has not been applied widely in all hospitals. Moreover, IHC has been proposed to be a pre-screening tool because of its wide application in clinics. Since the low expression of ALK protein, the sensitivity and specificity of ALK antibody are the keys to the success of IHC screening. Therefore, we compared different antibodies to find the best one for IHC detection. MATERIALS AND METHODS: We evaluated ALK expression by four different ALK antibodies: clone D5F3 (Ventana), clone D5F3 (CST), clone 1A4/1H7 (OriGene Tech.), and clone 5A4 (Abcam) based on manual IHC in a cohort of 60 NSCLCs. The results were compared with those from automated IHC (clone D5F3, Ventana). All cases were evaluated independently by ALK FISH. RESULTS: 32 ALK-positive and 28 ALK-negative NSCLCs were identified by automated IHC (D5F3, Ventana) and FISH analysis. Based on conventional manual IHC, the sensitivity of four antibodies-D5F3 (Ventana), D5F3 (CST), 1A4/1H7 (OriGene Tech.), and 5A4 (Abcam)-was 93.8%, 84.4%, 93.8%, and 56.3%, respectively. Their specificities and positive predictive values were 100%. The percentage of strong-moderate staining was 65.6%, 62.5%, 68.8%, and 21.9%, respectively. Compared with automated IHC (D5F3, Ventana), each staining concordance was 96.7%, 91.7%, 96.7%, and 76.7%, respectively, and each presented staining heterogeneity (weak-moderate-strong intensity). CONCLUSION: These data indicated that manual IHC with a more reliable ALK antibody might provide an effective strategy for screening ALK gene rearrangements in all NSCLC patients, followed by confirmatory FISH analysis in IHC-positive cases. CI - Copyright (c) 2015 Elsevier Ireland Ltd. All rights reserved. FAU - Shen, Qin AU - Shen Q AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. Electronic address: qinshen2005@163.com. FAU - Wang, Xuan AU - Wang X AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. FAU - Yu, Bo AU - Yu B AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. FAU - Shi, Shanshan AU - Shi S AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. FAU - Liu, Biao AU - Liu B AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. FAU - Wang, Yanfen AU - Wang Y AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. FAU - Xia, Qiuyuan AU - Xia Q AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. FAU - Rao, Qiu AU - Rao Q AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. FAU - Zhou, Xiaojun AU - Zhou X AD - Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. Electronic address: zhouxj3456@126.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20151021 PL - Ireland TA - Lung Cancer JT - Lung cancer (Amsterdam, Netherlands) JID - 8800805 RN - EC 2.7.10.1 (ALK protein, human) RN - EC 2.7.10.1 (Anaplastic Lymphoma Kinase) RN - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases) SB - IM MH - Anaplastic Lymphoma Kinase MH - Carcinoma, Non-Small-Cell Lung/diagnosis/*genetics/*metabolism MH - Female MH - *Gene Rearrangement MH - Humans MH - Immunohistochemistry MH - In Situ Hybridization, Fluorescence MH - Lung Neoplasms/diagnosis/*genetics/*metabolism MH - Male MH - Receptor Protein-Tyrosine Kinases/*genetics/*metabolism OTO - NOTNLM OT - Anaplastic lymphoma kinase (ALK) gene rearrangement OT - Antibody 1A4/1H7 OT - Immunohistochemistry (IHC) OT - Non-small cell lung cancer (NSCLC) EDAT- 2015/10/20 06:00 MHDA- 2016/10/14 06:00 CRDT- 2015/10/20 06:00 PHST- 2015/08/22 00:00 [received] PHST- 2015/09/26 00:00 [revised] PHST- 2015/10/04 00:00 [accepted] PHST- 2015/10/20 06:00 [entrez] PHST- 2015/10/20 06:00 [pubmed] PHST- 2016/10/14 06:00 [medline] AID - S0169-5002(15)30071-4 [pii] AID - 10.1016/j.lungcan.2015.10.002 [doi] PST - ppublish SO - Lung Cancer. 2015 Dec;90(3):492-8. doi: 10.1016/j.lungcan.2015.10.002. Epub 2015 Oct 21.