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

International Journal of Molecular Sciences

Gernot Kriegshäuser²  Gerhild Fabjani¹  Barbara Ziegler¹  Sabine Zཬhbauer-Müller⁴  Adelheid End³ 

[1] Molecular Oncology Group, Department of Obstetrics and Gynecology, Medical University Vienna, 1090 Vienna, Austria; E-Mails:;ViennaLab Diagnostics GmbH, 1120 Vienna, Austria; E-Mail:;Department of Cardiothoracic Surgery, Medical University of Vienna, 1090 Vienna, Austria; E-Mail:;Division of Oncology, Department of Internal Medicine I, Medical University of Vienna, 1090 Vienna, Austria; E-Mail:

关键词: non-small cell lung cancer;    KRAS;    mutation detection;    biochip hybridization;   

DOI  :  10.3390/ijms12128530

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2011 by the authors; licensee MDPI, Basel, Switzerland.