CAS OpenIR  > 中科院上海应用物理研究所2004-2010年
Ligase-based multiple DNA analysis by using an electrochemical sensor array
Wan, Y; Zhang, J; Liu, G; Pan, D; Wang, LH(王丽华); Song, SP(宋世平); Fan, CH(樊春海); Song, SP (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China
2009
Source PublicationBIOSENSORS & BIOELECTRONICS
ISSN0956-5663
Volume24Issue:5Pages:1209
AbstractWe herein report an electrochemical biosensor for the sequence-specific detection of DNA with high discrimination ability for single-nucleotide polymorphisms (SNPs). This DNA sensor was constructed by a pair of flanking probes that "sandwiched" the target. A 16-electrode electrochemical sensor array was employed, each having one individual DNA capture probe immobilized at gold electrodes via gold-thiol chemistry. By coupling with a biotin-tagged detection probe, we were able to detect multiple DNA targets with a single array. In order to realize SNP detection, a ligase-based approach was employed. In this method, both the capture probe and the detection probe were in tandem upon being hybridized with the target. Importantly, we employed a ligase that specifically could ligate tandem sequences only in the absence of mismatches. As a result, when both probes were complementary to the target, they were ligated in the presence of the ligase, thus being retained at the surface during the subsequent stringent washing steps. In contrast, if there existed 1-base mismatch, which could be efficiently recognized by the ligase, the detection probe was not ligated and subsequently washed away. A conjugate of avidin-horseradish peroxidase was then attached to the biotin label at the end of the detection probe via the biotin-avidin bridge. We then electrochemically interrogated the electrical current for the peroxidase-catalyzed reduction of hydrogen peroxide. We demonstrated that the electrochemical signal for the wild-type DNA was significantly larger than that for the sequence harboring the SNP. (c) 2008 Elsevier B.V. All rights reserved.
Subject AreaBiophysics ; Biotechnology & Applied Microbiology ; Chemistry ; Electrochemistry ; Science & Technology - Other Topics
Indexed BySCI
Language英语
Funding Project应物所项目组
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/7767
Collection中科院上海应用物理研究所2004-2010年
Corresponding AuthorSong, SP (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China
Recommended Citation
GB/T 7714
Wan, Y,Zhang, J,Liu, G,et al. Ligase-based multiple DNA analysis by using an electrochemical sensor array[J]. BIOSENSORS & BIOELECTRONICS,2009,24(5):1209.
APA Wan, Y.,Zhang, J.,Liu, G.,Pan, D.,Wang, LH.,...&Song, SP .(2009).Ligase-based multiple DNA analysis by using an electrochemical sensor array.BIOSENSORS & BIOELECTRONICS,24(5),1209.
MLA Wan, Y,et al."Ligase-based multiple DNA analysis by using an electrochemical sensor array".BIOSENSORS & BIOELECTRONICS 24.5(2009):1209.
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