Detection of EGFR-TK domain-activating mutations in NSCLC with generic PCR-based methods

Applied Immunohistochemistry & Molecular Morphology : AIMM
Rajendra B ShahiErik Teugels

Abstract

Somatic mutations in the epidermal growth factor receptor-tyrosine kinase (EGFR-TK) domain of non-small cell lung cancer (NSCLC) influence the responsiveness of these tumors to EGFR-TK inhibitors, indicating their usefulness as a predictive molecular marker. However, for mutation analysis, the amount of clinical material available from NSCLC patients is often very limited, suboptimally preserved, and composed of both normal and tumor cells. As a consequence, the total amount of recovered DNA is frequently very limited, with mutant alleles being often strongly underrepresented, and thus requiring highly sensitive methods for the detection of mutations. In the present study, EGFR mutation screening was performed on 210 NSCLC clinical samples by heminested polymerase chain reaction (PCR), followed by denaturing gradient gel electrophoresis (DGGE). Candidate mutations were further characterized by sequencing. Seventeen different types of pathogenic EGFR-TK domain mutations were detected in 55 of the 210 samples (26%). We reanalyzed 149 of the 155 samples in which no mutation was found by real-time PCR for the presence of recurrent exon 21 and exon 19 mutations using peptide nucleic acid probes in the PCR mix to increase sensitivity...Continue Reading

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Citations

Dec 9, 2016·Expert Review of Respiratory Medicine·Ting-Hui WuJames Chih-Hsin Yang
May 8, 2018·The Oncologist·Julia Lai-KwonJeremy Lewin
Dec 15, 2020·Journal of Applied Clinical Medical Physics·Yutao DangYi Zhang

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Methods Mentioned

BETA
electrophoresis
PCR
biopsy

Software Mentioned

LightCycler 480

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