Diamond: Immunohistochemistry versus sequencing in EGFR analysis of lung adenocarcinomas

Moira Ragazzi, Ione Tamagnini, Alessandra Bisagni, Alberto Cavazza, Maria Pagano, Licia Baldi, Corrado Boni, Flavia Cantile, Fausto Barbieri, Davide Nicoli, Giuliana Sartori, Dario De Biase, Giorgio Gardini, Giulio Rossi

Research output: Contribution to journalArticlepeer-review


Aims Identification of epidermal growth factor receptor (EGFR) mutations in lung adenocarcinomas is the single most important predictor of clinical response and outcome using EGFR tyrosine kinase inhibitors (TKIs). EGFR E746-A750del and L858R mutations are the most common gene alterations, also predicting the best clinical response to TKIs. We evaluated the accuracy of EGFR mutation-specific antibodies in a large cohort of lung adenocarcinomas, with different molecular settings and types of tissue samples. Methods 300 lung adenocarcinomas diagnosed on cytology (48 cell blocks), biopsy (157 cases) and surgical resections (95 cases) were selected. All cases were investigated for EGFR by sequencing and two mutationspecific antibodies (clone 6B6 for E746-A750del; clone 43B2 for L858R) were tested using an automated immunostainer. Discordant results were investigated by next-generation sequencing (NGS). Results Overall sensitivity and specificity of mutantspecific antibodies were 58.6% and 98.0%, respectively, and they increased up to 84% and 100% if only tumours harbouring E746-A750del were considered. In 13 discordant cases, NGS confirmed immunohistochemistry results in eight samples. Conclusions The EGFR mutation-specific antibodies have a fair/good sensitivity and good/high specificity in identifying classic mutations, but they cannot replace molecular tests. The antibodies work equally well on biopsies and cell blocks, possibly permitting a rapid screening in cases with poor material.

Original languageEnglish
Pages (from-to)440-447
Number of pages8
JournalJournal of Clinical Pathology
Issue number5
Publication statusPublished - May 1 2016

ASJC Scopus subject areas

  • Pathology and Forensic Medicine


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