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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Molecular Diversity Preservation International (MDPI)
2011
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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. |
format | Online Article Text |
id | pubmed-3257086 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Molecular Diversity Preservation International (MDPI) |
record_format | MEDLINE/PubMed |
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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