Multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers  

作　　者：XING BaoLing SONG JinZhao GE SuMei TANG ZhengHua LIU MengLu YANG Qiong Lü FengTing LIU LiBing WANG Shu 

机构地区：[1]Department of Pathology, Changzhou Wommen and Children Health Hospital [2]Key Laboratory of Organic Solids, Institute of Chemistry, Chinese Academy of Sciences

出　　处：《Chinese Science Bulletin》2013年第8期873-878,共6页

基　　金：supported by the National Natural Science Foundation of China (90913014, 21021091)

摘　　要：The mutation detections of KRAS and BRAF genes are of significant importance to predict the responses to anti-cancer therapy and develop new drugs. In this paper, we developed a multi-step fluorescence resonance energy transfer (FRET) assay for multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers (CCP). The newly established detection system could detect as low as 2% mutant DNAs in DNA admixtures. By triggering the emission intensity change of CCP and the dyes labeled in the DNA, four possible statuses (three mutations and one wildtype) can be differentiated in one extension reaction. The detection efficiency of this new method in clinical molecular diagnosis was validated by determining KRAS and BRAF mutations of 51 formalin-fixed paraffin-embedded (FFPE) ovary tissue samples. Furthermore, the result of the CCP-based multi-step FRET assay can be directly visualized under UV light so that no expensive instruments and technical expertise are needed. Thus, the assay provides a sensitive, reliable, cost-effective and simple method for the detection of disease-related gene mutations.The mutation detections of KRAS and BRAF genes are of significant importance to predict the responses to anti-cancer therapy and develop new drugs. In this paper, we developed a multi-step fluorescence resonance energy transfer （FRET） assay for multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers （CCP）. The newly established detection system could detect as low as 2% mutant DNAs in DNA admixtures. By triggering the emission intensity change of CCP and the dyes labeled in the DNA, four possible statuses （three mutations and one wildtype） can be differentiated in one extension reaction. The detection efficiency of this new method in clinical molecular diagnosis was validated by determining KRAS and BRAF mutations of 51 formalin-fixed paraffin-embedded （FFPE） ovary tissue samples. Furthermore, the result of the CCP-based multi-step FRET assay can be directly visualized under UV light so that no expensive instruments and technical expertise are needed. Thus, the assay provides a sensitive, reliable, cost-effective and simple method for the detection of disease-related gene mutations.

关 键 词：基因突变检测 共轭聚合物 阳离子 多重 荧光共振能量转移 中国共产党 DNA FRET 

分 类 号：R730.5[医药卫生—肿瘤]