Comparison of Targeted Next-Generation Sequencing (NGS) and Real-Time PCR in the Detection ofEGFR,KRAS,andBRAFMutations on Formalin-Fixed, Paraffin-Embedded Tumor Material of Non-Small Cell Lung Carcinoma-Superiority of NGS

Autor: Katja Tuononen, Satu Mäki-Nevala, Aino I. Telaranta-Keerie, Sari Hannula, Virinder Kaur Sarhadi, Mikko Juhani Rönty, Kaisa Salmenkivi, Aija Knuuttila, Sonja Lagström, Pekka Ellonen, Aino Wirtanen, Jenny M. Andrews, Sakari Knuutila
Rok vydání: 2013
Předmět:
Adult
Male
Proto-Oncogene Proteins B-raf
Cancer Research
Lung Neoplasms
medicine.drug_class
DNA Mutational Analysis
Biology
Gene mutation
Real-Time Polymerase Chain Reaction
medicine.disease_cause
DNA sequencing
Tyrosine-kinase inhibitor
law.invention
Proto-Oncogene Proteins p21(ras)
Fixatives
03 medical and health sciences
0302 clinical medicine
INDEL Mutation
law
Carcinoma
Non-Small-Cell Lung
Formaldehyde
Proto-Oncogene Proteins
Genetics
Carcinoma
medicine
Humans
Genetic Testing
Genetic Association Studies
Polymerase chain reaction
Aged
030304 developmental biology
Aged
80 and over
0303 health sciences
Paraffin Embedding
High-Throughput Nucleotide Sequencing
Cancer
Middle Aged
medicine.disease
Molecular biology
3. Good health
ErbB Receptors
Real-time polymerase chain reaction
Molecular Diagnostic Techniques
030220 oncology & carcinogenesis
ras Proteins
Cancer research
Female
KRAS
Zdroj: Genes, Chromosomes and Cancer. 52:503-511
ISSN: 1045-2257
Popis: The development of tyrosine kinase inhibitor treatments has made it important to test cancer patients for clinically significant gene mutations that influence the benefit of treatment. Targeted next-generation sequencing (NGS) provides a promising method for diagnostic purposes by enabling the simultaneous detection of multiple mutations in various genes in a single test. The aim of our study was to screen EGFR, KRAS, and BRAF mutations by targeted NGS and commonly used real-time polymerase chain reaction (PCR) methods to evaluate the feasibility of targeted NGS for the detection of the mutations. Furthermore, we aimed to identify potential novel mutations by targeted NGS. We analyzed formalin-fixed, paraffin-embedded (FFPE) tumor tissue specimens from 81 non-small cell lung carcinoma patients. We observed a significant concordance (from 96.3 to 100%) of the EGFR, KRAS, and BRAF mutation detection results between targeted NGS and real-time PCR. Moreover, targeted NGS revealed seven nonsynonymous single-nucleotide variations and one insertion-deletion variation in EGFR not detectable by the real-time PCR methods. The potential clinical significance of these variants requires elucidation in future studies. Our results support the use of targeted NGS in the screening of EGFR, KRAS, and BRAF mutations in FFPE tissue material.
Databáze: OpenAIRE
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