title: Detection of KRAS mutations using double-stranded toehold-exchange probes.
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.
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China; University of Chinese Academy of Sciences, Beijing 100039, China.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Department of Gastroenterology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Tianle
authors.initials: T
authors.lastname: Ma
authors.affiliation: INSERM U823, Institut Albert Bonniot, Grenoble, France; Université Joseph Fourier UJF, Grenoble, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Coll
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China.
authors.forename: Fangming
authors.initials: F
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China.
authors.forename: Honglian
authors.initials: H
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China.
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China; University of Chinese Academy of Sciences, Beijing 100039, China.
authors.forename: Zhishuo
authors.initials: Z
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China.
authors.forename: Qinghui
authors.initials: Q
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China. Electronic address: hjmao@mail.sim.ac.cn.
authors.forename: Hongju
authors.initials: H
authors.lastname: Mao
authors.affiliation: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China. Electronic address: jlzhao@mail.sim.ac.cn.
authors.forename: Jianlong
authors.initials: J
authors.lastname: Zhao
mesh_terms: D000483:Alleles; D015374:Biosensing Techniques; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D005453:Fluorescence; D006801:Humans; D009154:Mutation; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics / Q000302:isolation & purification*
chemical_list: C117307:KRAS protein, human; D004247:DNA; D016283:Proto-Oncogene Proteins p21(ras)
keywords: DNA; KRAS; Mutation; Toehold-exchange Probe