Determination of EGFR gene somatic mutations in tissues and plasma of patients with non-small cell lung cancer


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Activating mutations in the EGFR gene influence cell proliferation, angiogenesis, and increases metastatic ability of non-small cell lung cancer (NSCLC) cells; they have a significant impact on the choice of medical therapy of NSCLC. The use of targeted therapy with tyrosine kinase inhibitors requires performance of appropriate genetic tests in NSCLC patients. The aim of this study was to develop a real-time PCRbased diagnostic test-system for rapid and cost-effective analysis of EGFR mutations in paraffin blocks and plasma and to perform comparative estimation of diagnostic characteristics features of real-time wild type blocking PCR and digital PCR. The study included 156 patients with different degrees of lung adenocarcinoma differentiation. A simple and efficient real-time PCR-based method for detection of L858R activating mutation and del19 deletion in the EGFR gene in DNA isolated from paraffin blocks or blood has been developed. The test system for EGFR mutations has been validated using 411 samples of paraffin blocks. The proposed system demonstrated high efficiency for DNA testing from paraffin blocks: a concordance with results of testing by means a Therascreen® EGFR RGQ PCR Kit (Qiagen, Germany) was 100%. Applicability of this test system has been also demonstrated for detection of mutations in plasma.

作者简介

O. Brovkina

Federal Research and Clinical Center for Specialized Types of Medical Care and Technologies

编辑信件的主要联系方式.
Email: brov.olia@gmail.com
俄罗斯联邦, Orekhovyi bulv. 28, Moscow, 115682

M. Gordiev

Tatarstan Cancer Center

Email: brov.olia@gmail.com
俄罗斯联邦, Kazan

A. Toropovskiy

LLC “TestGene,”

Email: brov.olia@gmail.com
俄罗斯联邦, Ulianovsk

D. Khodyrev

Federal Research and Clinical Center for Specialized Types of Medical Care and Technologies

Email: brov.olia@gmail.com
俄罗斯联邦, Orekhovyi bulv. 28, Moscow, 115682

R. Enikeev

Tatarstan Cancer Center

Email: brov.olia@gmail.com
俄罗斯联邦, Kazan

O. Gusev

Kazan Federal University; RIKEN

Email: brov.olia@gmail.com
俄罗斯联邦, Kazan; Yokohama

L. Shigapova

Kazan Federal University

Email: brov.olia@gmail.com
俄罗斯联邦, Kazan

A. Nikitin

Federal Research and Clinical Center for Specialized Types of Medical Care and Technologies

Email: brov.olia@gmail.com
俄罗斯联邦, Orekhovyi bulv. 28, Moscow, 115682

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