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Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer

This study is aimed at evaluating the potential of a biochip assay to sensitively detect KRAS mutation in DNA from non-small cell lung cancer (NSCLC) tissue samples. The assay covers 10 mutations in codons 12 and 13 of the KRAS gene, and is based on mutant-enriched PCR followed by reverse-hybridizat...

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Autores principales: Kriegshäuser, Gernot, Fabjani, Gerhild, Ziegler, Barbara, Zöchbauer-Müller, Sabine, End, Adelheid, Zeillinger, Robert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3257086/
https://www.ncbi.nlm.nih.gov/pubmed/22272089
http://dx.doi.org/10.3390/ijms12128530
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author Kriegshäuser, Gernot
Fabjani, Gerhild
Ziegler, Barbara
Zöchbauer-Müller, Sabine
End, Adelheid
Zeillinger, Robert
author_facet Kriegshäuser, Gernot
Fabjani, Gerhild
Ziegler, Barbara
Zöchbauer-Müller, Sabine
End, Adelheid
Zeillinger, Robert
author_sort Kriegshäuser, Gernot
collection PubMed
description This study is aimed at evaluating the potential of a biochip assay to sensitively detect KRAS mutation in DNA from non-small cell lung cancer (NSCLC) tissue samples. The assay covers 10 mutations in codons 12 and 13 of the KRAS gene, and is based on mutant-enriched PCR followed by reverse-hybridization of biotinylated amplification products to an array of sequence-specific probes immobilized on the tip of a rectangular plastic stick (biochip). Biochip hybridization identified 17 (21%) samples to carry a KRAS mutation of which 16 (33%) were adenocarcinomas and 1 (3%) was a squamous cell carcinoma. All mutations were confirmed by DNA sequencing. Using 10 ng of starting DNA, the biochip assay demonstrated a detection limit of 1% mutant sequence in a background of wild-type DNA. Our results suggest that the biochip assay is a sensitive alternative to protocols currently in use for KRAS mutation testing on limited quantity samples.
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spelling pubmed-32570862012-01-23 Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer Kriegshäuser, Gernot Fabjani, Gerhild Ziegler, Barbara Zöchbauer-Müller, Sabine End, Adelheid Zeillinger, Robert Int J Mol Sci Article This study is aimed at evaluating the potential of a biochip assay to sensitively detect KRAS mutation in DNA from non-small cell lung cancer (NSCLC) tissue samples. The assay covers 10 mutations in codons 12 and 13 of the KRAS gene, and is based on mutant-enriched PCR followed by reverse-hybridization of biotinylated amplification products to an array of sequence-specific probes immobilized on the tip of a rectangular plastic stick (biochip). Biochip hybridization identified 17 (21%) samples to carry a KRAS mutation of which 16 (33%) were adenocarcinomas and 1 (3%) was a squamous cell carcinoma. All mutations were confirmed by DNA sequencing. Using 10 ng of starting DNA, the biochip assay demonstrated a detection limit of 1% mutant sequence in a background of wild-type DNA. Our results suggest that the biochip assay is a sensitive alternative to protocols currently in use for KRAS mutation testing on limited quantity samples. Molecular Diversity Preservation International (MDPI) 2011-11-29 /pmc/articles/PMC3257086/ /pubmed/22272089 http://dx.doi.org/10.3390/ijms12128530 Text en © 2011 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Kriegshäuser, Gernot
Fabjani, Gerhild
Ziegler, Barbara
Zöchbauer-Müller, Sabine
End, Adelheid
Zeillinger, Robert
Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer
title Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer
title_full Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer
title_fullStr Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer
title_full_unstemmed Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer
title_short Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer
title_sort biochip-based detection of kras mutation in non-small cell lung cancer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3257086/
https://www.ncbi.nlm.nih.gov/pubmed/22272089
http://dx.doi.org/10.3390/ijms12128530
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