Qin Shen1, Xuan Wang2, Bo Yu2, Shanshan Shi2, Biao Liu2, Yanfen Wang2, Qiuyuan Xia2, Qiu Rao2, Xiaojun Zhou3. 1. 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. 2. Department of Pathology, Jinling Hospital, Clinical Medical School of Southern Medical University, 305 East Zhongshan Road, Nanjing 210002, PR China. 3. 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.
Abstract
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.
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 NSCLCpatients, followed by confirmatory FISH analysis in IHC-positive cases.
Authors: Francesco Facchinetti; Marcello Tiseo; Massimo Di Maio; Paolo Graziano; Emilio Bria; Giulio Rossi; Silvia Novello Journal: Transl Lung Cancer Res Date: 2016-06
Authors: Jerzy Lasota; Małgorzata Chłopek; Bartosz Wasąg; Artur Kowalik; Jason Christiansen; Jennifer Lamoureux; Alina Kuźniacka; Anna Felisiak-Gołąbek; Yalan Liu; Tiffany Ashley R Reyes; Rishabh Saha; Abbas Agaimy; Kristyna Behenska; Wojciech Biernat; Laura Cattaneo; Giovanni Centonze; Ondrej Daum; Magdalena Daumova; Paweł Domagała; Ireneusz Dziuba; Carol E Geppert; Stanisław Góźdź; Anna Nasierowska-Guttmejer; Agnieszka Hałoń; Arndt Hartmann; Shingo Inaguma; Ewa Iżycka-Świeszewska; Maciej Kaczorowski; Małgorzata Kołos; Janusz Kopczyński; Michal Michal; Massimo Milione; Krzysztof Okoń; Rafał Pęksa; Michał Pyzlak; Janusz Ryś; Piotr Waloszczyk; Jaroslaw Wejman; Markku Miettinen Journal: Am J Surg Pathol Date: 2020-09 Impact factor: 6.298