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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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.