Detection of KRAS mutations using double-stranded toehold-exchange probes

Biosensors & Bioelectronics
Zhenhua WuJianlong Zhao

Abstract

Detection of KRAS mutations in cancer tissues is immensely valuable for the identification of personalized genotype-based therapy. Here, we employed a double-stranded toehold-exchange probe, which is labeled with fluorescent molecules (FAM) and quenchers (Dabcyl), to detect KRAS mutations in cancer tissues. This probe was able to differentiate the intended mutation in a sample containing as little as 5% mutant alleles in a background of wild-type DNA. This probe also performed robustly at a wide range of conditions, for examples, from 4 °C to 37 °C, from 200 mM Na(+) to 1M Na(+), and from 200 mM K(+) to 500 mM K(+). Furthermore, we validated the practicality of this probe in a clinical setting using 8 pairs of cancer tissue samples and their NT (corresponding adjacent nontumorous tissue) samples. All the results generated from the probe detection agreed with those from direct sequencing. Combining features of extreme high specificity and robustness, this probe is a valuable tool for reliable diagnosis of cancer-related mutations.

References

Aug 26, 1988·Science·U LandegrenL Hood
Jul 13, 2000·Proceedings of the National Academy of Sciences of the United States of America·J G HallV I Lyamichev
Jun 28, 2005·Nucleic Acids Research·Nicholas R Markham, Michael Zuker
Apr 16, 2009·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Jian YuMichael J Comb
Mar 10, 2011·International Journal of Cancer. Journal International Du Cancer·Sascha TierlingJörn Walter
Feb 23, 2012·Nature Chemistry·David Yu ZhangPeng Yin
Jul 20, 2012·Diseases of the Colon and Rectum·JiaoJiao RenYaShuang Zhao
Aug 18, 2012·Cancer Epidemiology, Biomarkers & Prevention : a Publication of the American Association for Cancer Research, Cosponsored by the American Society of Preventive Oncology·Amanda I PhippsPolly A Newcomb
May 8, 2013·The American Journal of Gastroenterology·Quirinus J M VoorhamSilvia Sanduleanu
May 11, 2013·British Journal of Cancer·V EklöfR Palmqvist
Jan 9, 2015·Biosensors & Bioelectronics·Yong ShinMi Kyoung Park

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