PMID- 24667320 OWN - NLM STAT- MEDLINE DCOM- 20141230 LR - 20220310 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 9 IP - 3 DP - 2014 TI - Accurate and economical detection of ALK positive lung adenocarcinoma with semiquantitative immunohistochemical screening. PG - e92828 LID - 10.1371/journal.pone.0092828 [doi] LID - e92828 AB - Right detection of anaplastic lymphoma kinase (ALK) gene rearrangement is pivotal to selection of patients with lung adenocarcinoma for ALK-targeted therapy. We explored the potential of combination of immunohistochemistry (IHC) screening and fluorescence in situ hybridization (FISH) as an affordable practice. We analyzed 410 unselected lung adenocarcinomas by ALK IHC (D5F3 clone) and FISH. Some equivocal cases were further analyzed by RT-PCR. The EGFR mutation was detected by pyrosequencing assay. In total 368 cases which got all IHC, FISH, EGFR mutation results were eligible for analysis. Cases were evaluated as IHC score 3+ (n = 26), score 2+ (n = 9), score 1+ (n = 51), and score 0 (n = 282), respectively. 23 of 26 IHC 3+ and 5 of 9 IHC 2+ cases were FISH positive, whereas 3 of 26 IHC 3+, 4 of 9 IHC 2+ and all 333 IHC 1+/0 cases were FISH negative. If considering FISH as the standard, the sensitivity and specificity of ALK IHC 3+/2+ as ALK positive were 100% and 97.9%, respectively. Three IHC 3+ cases reported as FISH "negative" were actually ALK positive confirmed by ALK RT-PCR or re-detected. Based on the final classify, ALK IHC 3+/2+ was 100% sensitive and 98.8% specific. However, FISH was 90.3% sensitive and 100% specific. IHC 2+ was regarded as equivocal and need to be confirmed by FISH or RT-PCR. In the 368 cases, 8.4% cases had ALK positive, 52.2% cases had EGFR mutation, and only one case had a coexisting. Manually semiquantitative ALK IHC (primary antibody D5F3 coupled with secondary DAKO Envision system) used as the initial screening combined with auxiliary FISH confirmation is a reliable, economical approach to identify ALK positive lung adenocarcinoma. The IHC can find some ALK positive cases which would be missed by FISH only. FAU - Zhou, Jianya AU - Zhou J AD - Department of Respiratory Disease, Thoracic Disease Center, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Zhao, Jing AU - Zhao J AD - Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Sun, Ke AU - Sun K AD - Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Wang, Bo AU - Wang B AD - Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Wang, Lijun AU - Wang L AD - Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Chen, Xi AU - Chen X AD - Department of Respiratory Disease, Thoracic Disease Center, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Zheng, Jing AU - Zheng J AD - Department of Respiratory Disease, Thoracic Disease Center, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - You, Qihan AU - You Q AD - Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Wang, Xiaoling AU - Wang X AD - Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Ding, Wei AU - Ding W AD - Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. FAU - Zhou, Jianying AU - Zhou J AD - Department of Respiratory Disease, Thoracic Disease Center, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. LA - eng PT - Clinical Trial PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140325 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - EC 2.7.10.1 (ALK protein, human) RN - EC 2.7.10.1 (Anaplastic Lymphoma Kinase) RN - EC 2.7.10.1 (EGFR protein, human) RN - EC 2.7.10.1 (ErbB Receptors) RN - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases) SB - IM MH - *Adenocarcinoma/economics/metabolism/pathology MH - Anaplastic Lymphoma Kinase MH - ErbB Receptors/genetics/metabolism MH - Female MH - Humans MH - *Immunohistochemistry/economics/methods MH - In Situ Hybridization, Fluorescence/economics/methods MH - *Lung Neoplasms/economics/metabolism/pathology MH - Male MH - Mutation MH - *Receptor Protein-Tyrosine Kinases PMC - PMC3965450 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2014/03/29 06:00 MHDA- 2014/12/31 06:00 PMCR- 2014/03/25 CRDT- 2014/03/27 06:00 PHST- 2013/12/03 00:00 [received] PHST- 2014/02/26 00:00 [accepted] PHST- 2014/03/27 06:00 [entrez] PHST- 2014/03/29 06:00 [pubmed] PHST- 2014/12/31 06:00 [medline] PHST- 2014/03/25 00:00 [pmc-release] AID - PONE-D-13-50660 [pii] AID - 10.1371/journal.pone.0092828 [doi] PST - epublish SO - PLoS One. 2014 Mar 25;9(3):e92828. doi: 10.1371/journal.pone.0092828. eCollection 2014.