Qiushi Wang1, Lianhua Zhao1, Xin Yang1, Shirong Wei1, Ying Zeng1, Chengyi Mao1, Li Lin1, Ping Fu1, Liang Lyu1, Zengpeng Li1, Hualiang Xiao2. 1. Department of Pathology, Daping Hospital and Research Institute of Surgery, Third Military Medical University, 10# Changjiangzhilu, Daping, Yuzhong District, Chongqing 400042, China. 2. Department of Pathology, Daping Hospital and Research Institute of Surgery, Third Military Medical University, 10# Changjiangzhilu, Daping, Yuzhong District, Chongqing 400042, China. Electronic address: dpbl_xhl@126.com.
Abstract
OBJECTIVES: The rearrangement of echinoderm microtubule-associated protein-like 4-analplastic lymphoma kinase (EML4-ALK) in non-small cell lung cancer (NSCLC) cells might be a promising therapeutic target. However, the low positive rate seeks a reliable and cost-effective method for ALK rearrangement prescreening. This study aimed to evaluate the application of a novel primary antibody 1A4 for routine ALK immunohistochemistry (IHC) test. MATERIALS AND METHODS: Primary antibody 1A4 and D5F3 were used for the screening of 595 formalin-fixed, paraffin-embedded tissues of consecutive patients with lung adenocarcinoma for ALK-positive candidates. Ventana detection system and fluorescence in-situ hybridization (FISH) were used as reference methods. RESULTS: Among 595 cases, the protein expression statuses of 1A4 were 3+ (18), 2+ (50), 1+ (153), and 0+ (374), and those of D5F3 were 3+ (17), 2+ (18), 1+ (20), and 0+ (540). Ventana detection system and FISH test results were successfully obtained from 482 cases. A total of 298 specimens with 1A4 (-) showed 100% concordance with standard FISH results. All 58 FISH (+) cases were identified by antibody 1A4. Meanwhile, 14 and 5 were missed by antibody D5F3 with routine IHC and Ventana system, respectively. 1A4 with routine IHC had better sensitivity (100%, 75.9%, and 91.4%, respectively), but lower specificity (70.3%, 99.8%, and 100%, respectively), than D5F3 with routine IHC and Ventana system. CONCLUSION: The novel antibody 1A4 used as a prescreening method may help to reduce the false-negative rearranged ALK status if FISH or reverse transcription polymerase chain reaction results were used for validation.
OBJECTIVES: The rearrangement of echinoderm microtubule-associated protein-like 4-analplastic lymphoma kinase (EML4-ALK) in non-small cell lung cancer (NSCLC) cells might be a promising therapeutic target. However, the low positive rate seeks a reliable and cost-effective method for ALK rearrangement prescreening. This study aimed to evaluate the application of a novel primary antibody 1A4 for routine ALK immunohistochemistry (IHC) test. MATERIALS AND METHODS: Primary antibody 1A4 and D5F3 were used for the screening of 595 formalin-fixed, paraffin-embedded tissues of consecutive patients with lung adenocarcinoma for ALK-positive candidates. Ventana detection system and fluorescence in-situ hybridization (FISH) were used as reference methods. RESULTS: Among 595 cases, the protein expression statuses of 1A4 were 3+ (18), 2+ (50), 1+ (153), and 0+ (374), and those of D5F3 were 3+ (17), 2+ (18), 1+ (20), and 0+ (540). Ventana detection system and FISH test results were successfully obtained from 482 cases. A total of 298 specimens with 1A4 (-) showed 100% concordance with standard FISH results. All 58 FISH (+) cases were identified by antibody 1A4. Meanwhile, 14 and 5 were missed by antibody D5F3 with routine IHC and Ventana system, respectively. 1A4 with routine IHC had better sensitivity (100%, 75.9%, and 91.4%, respectively), but lower specificity (70.3%, 99.8%, and 100%, respectively), than D5F3 with routine IHC and Ventana system. CONCLUSION: The novel antibody 1A4 used as a prescreening method may help to reduce the false-negative rearranged ALK status if FISH or reverse transcription polymerase chain reaction results were used for validation.
Authors: Cleo Keppens; Véronique Tack; Nils 't Hart; Lien Tembuyser; Ales Ryska; Patrick Pauwels; Karen Zwaenepoel; Ed Schuuring; Florian Cabillic; Luigi Tornillo; Arne Warth; Wilko Weichert; Elisabeth Dequeker Journal: Oncotarget Date: 2018-04-17