Simultaneous detection of double gene-specific methylation loci based on hairpin probes tagged with electrochemical quantum dots barcodes
A novel label-free and PCR-free electrochemical method is demonstrated for simultaneous evaluation of double gene-specific methylation loci in a single-tube experiment without numerous hybridization and heating processes.The method composes three units: two electrochemical quantum dots tagged dual-functional hairpin probes (HP1-CdS and HP2-PbS) for methylation loci recognition,and capture probes modified magnetic beads (MB-CP) as isolation vehicle.A p53 gene fragment with twomethylation loci was selected as a target model.After pretreatment with sodium bisulfite,samples were simply incubated with HP1-PbS,HP2-CdS,and MB-CP conjugates.The resultant MBs were then isolated and stripping analyzed by square wave voltammetry (SWV).Fromthe SWV responses of the QDs barcodes hybridized on MBs,the methylation pattern and level of target gene fragment were clearly identified.The linear range was 5.0-125.0 nM with a limit of detection (LOD) of 1.3 nM for locus A which was tagged by PbS,and 0.5-125.0 nM with a LOD of 0.38 nM for locus B which was tagged by CdS.The method showed satisfactory accuracy and testing recovery.In comparisonwith the current methods,the proposed method is convenient and can evaluate two gene-specific methylation in a single-tube experiment within 2.5 h.With regard to the advantages of facility,low-cost,and ease of operation,the method can be a potential platform for identification of DNA methylation status in a simple way.
Gene-specific methylation Quantum dots barcodes Simultaneous quantification Electrochemistry
Yuzhi Xu Xiaoyu Gao Li Zhang Danping Chen Zong Dai Xiaoyong Zou
School of Chemistry and Chemical Engineering,Sun Yat-Sen University,Guangzhou 510275,PR China
国际会议
南京
英文
356-362
2016-12-23（万方平台首次上网日期，不代表论文的发表时间）