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

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Author(s): Gernot Kriegshäuser | Gerhild Fabjani | Barbara Ziegler | Sabine Zöchbauer-Müller | Adelheid End | Robert Zeillinger

Journal: International Journal of Molecular Sciences
ISSN 1422-0067

Volume: 12;
Issue: 12;
Start page: 8530;
Date: 2011;
Original page

Keywords: non-small cell lung cancer | KRAS | mutation detection | biochip hybridization

ABSTRACT
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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