Immunohistochemical detection of mutations in the epidermal growth factor receptor gene in lung adenocarcinomas using mutation-specific antibodies

Diagnostic Pathology

Ting Li¹  Ying Dong¹  Jing Zhao¹  Lin Nong¹  Haodong Xu²  Paul G Rothberg²  Todd S Laughlin²  Nufatt Leong²  Yun Bai¹  Yan Xiong¹ 

[1] Department of Pathology, Peking University First Hospital, 7 Xishiku Street, Xicheng District, Beijing, 100034, China;Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, 601 Elmwood Avenue, Rochester, NY, 14642, USA

关键词: Immunohistochemistry;    Mutation;    Epidermal growth factor receptor;    Lung adenocarcinoma;   

Others  :  807503
DOI  :  10.1186/1746-1596-8-27

 received in 2012-12-18, accepted in 2013-02-05,  发布年份 2013

【 摘 要 】

Background

The recent development of antibodies specific for the major hotspot mutations in the epidermal growth factor receptor (EGFR), L858R and E746_A750del, may provide an opportunity to use immunohistochemistry (IHC) as a screening test for EGFR gene mutations. This study was designed to optimize the IHC protocol and the criteria for interpretation of the results using DNA sequencing as the gold-standard.

Methods

Tumor sections from fifty lung adenocarcinoma specimens from Chinese patients were immunostained using L858R and E746_A750del-specific antibodies using three different antigen retrieval solutions, and the results were evaluated using three different sets of criteria. The same specimens were used for DNA purification and analysis of EGFR gene mutations.

Results

In this study the optimal buffer for antigen retrieval was EDTA (pH 8.0), and the optimal scoring method was to call positive results when there was moderate to strong staining of membrane and/or cytoplasm in >10% of the tumor cells. Using the optimized protocol, L858R-specific IHC showed a sensitivity of 81% and a specificity of 97%, and E746_A750del-specific IHC showed a sensitivity of 59% and a specificity of 100%, both compared with direct DNA analysis. Additionally, the mutant proteins as assessed by IHC showed a more homogeneous than heterogeneous pattern of expression.

Conclusions

Our data demonstrate that mutation-specific IHC, using optimized procedures, is a reliable prescreening test for detecting EGFR mutations in lung adenocarcinoma.

Virtual Slides

The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2059012601872392 webcite

【 授权许可】

   
2013 Xiong et al.; licensee BioMed Central Ltd.

【 图 表 】

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