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Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer
ViennaLab Diagnostics GmbH, 1120 Vienna, Austria
Molecular Oncology Group, Department of Obstetrics and Gynecology, Medical University Vienna, 1090 Vienna, Austria
Ludwig Boltzmann Gesellschaft, Cluster Translational Oncology, 1090 Vienna, Austria
Division of Oncology, Department of Internal Medicine I, Medical University of Vienna, 1090 Vienna, Austria
Department of Cardiothoracic Surgery, Medical University of Vienna, 1090 Vienna, Austria
* Author to whom correspondence should be addressed.
Received: 18 September 2011; in revised form: 17 November 2011 / Accepted: 17 November 2011 / Published: 29 November 2011
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.
Keywords: non-small cell lung cancer; KRAS; mutation detection; biochip hybridization
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Kriegshäuser, G.; Fabjani, G.; Ziegler, B.; Zöchbauer-Müller, S.; End, A.; Zeillinger, R. Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer. Int. J. Mol. Sci. 2011, 12, 8530-8538.
Kriegshäuser G, Fabjani G, Ziegler B, Zöchbauer-Müller S, End A, Zeillinger R. Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer. International Journal of Molecular Sciences. 2011; 12(12):8530-8538.
Kriegshäuser, Gernot; Fabjani, Gerhild; Ziegler, Barbara; Zöchbauer-Müller, Sabine; End, Adelheid; Zeillinger, Robert. 2011. "Biochip-Based Detection of KRAS Mutation in Non-Small Cell Lung Cancer." Int. J. Mol. Sci. 12, no. 12: 8530-8538.