Novel electrochemical real-time polymerase chain reaction utilizing silver ion aptamer-modified primer = 은 이온 압타머 수식 프라이머를 이용한 신개념 전기화학적 실시간 중합효소연쇄반응

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We herein describe a novel electrochemical real-time polymerase chain reaction (ERT-PCR) utilizing silver ion ($Ag^+$) aptamer-modified primer. In this work, we designed the primer to be extended by $Ag^+$ aptamer at the 5’ end, which would initially form a hairpin structure entrapping $Ag^+$. The $Ag^+$ in this bound state cannot actively participate in the redox reaction due to the low diffusivity, resulting in the low electrochemical signal. In the presence of target DNA, however, the $Ag^+$ is released from the aptamer as the opposite DNA strand is synthesized or as the $Ag^+$ aptamer-modified primer anneals to the extended template during the PCR process. Since the released free $Ag^+$ has much higher diffusivity than the bound one, the $Ag^+$-mediated electrochemical signal would increase in accordance with the number of PCR cycles. Based on this simple design principle, we successfully detected the Enterococcus faecium (E. faecium) genomic DNA (gDNA) down to $3.085 × 10^2 copies/\mul$ with excellent selectivity against non-target DNAs. This method has a great potential of serving as a key platform technology to advance an electrochemical genetic diagnostic system for a point-of-care testing (POCT) purpose.
Advisors
Park, Hyun Gyuresearcher박현규researcher
Description
한국과학기술원 :생명화학공학과,
Publisher
한국과학기술원
Issue Date
2019
Identifier
325007
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생명화학공학과, 2019.2,[iii, 33 p. :]

Keywords

ERT-PCR▼aaptamer-modified primer▼asilver ion▼aDNA▼abiosensor; 전기화학적 실시간 중합효소반응▼a압타머-수식 프라이머▼a은 이온▼aDNA▼a바이오센서

URI
http://hdl.handle.net/10203/266330
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=843731&flag=dissertation
Appears in Collection
CBE-Theses_Master(석사논문)
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