Aptamer-Based Rolling Circle Amplification:  A Platform for Electrochemical Detection of Protein

2007 ◽  
Vol 79 (19) ◽  
pp. 7492-7500 ◽  
Author(s):  
Long Zhou ◽  
Li-Juan Ou ◽  
Xia Chu ◽  
Guo-Li Shen ◽  
Ru-Qin Yu
Keyword(s):  
Electrochemical Detection ◽  
Rolling Circle Amplification ◽  
Rolling Circle

scholarly journals Rolling circle amplification with electrochemical detection assay for SARS-CoV-2 

2021 ◽  
Author(s):  
Thanyarat Chaibun ◽  
Jiratchaya Puenpa ◽  
Tatchanun Ngamdee ◽  
Nimaradee Boonapatcharoen ◽  
Pornpat Athamanolap ◽  
...  
Keyword(s):  
Electrochemical Detection ◽  
Rolling Circle Amplification ◽  
Differential Pulse ◽  
Rolling Circle ◽  
Redox Active ◽  
Detection Assay ◽  
Quantitative Results ◽  
Circular Template ◽  
Dna Template ◽  
Field Setting

Abstract This protocol describes the rolling circle amplification (RCA) and electrochemical detection of SARS-CoV-2. The procedure consists of 3 parts, which are the amplification, hybridization and detection steps. In the presence of RNA template, the circular DNA template will be ligated to produce a Padlock DNA, which serves as a template for amplification by phi29 DNA polymerase to produce RCA amplicons. The RCA amplicon is a concatemer containing multiple repeats of sequences that are complementary to the circular template. The RCA amplicons are hybridized with redox active probes and detected by electrochemical biosensor using differential pulse voltammetry (DPV). Due to its isothermal nature, RCA can be performed using a simple water bath or heating block. Overall, the whole assay takes approximately 45 min. The assay enables rapid, quantitative results to be obtained for detection of SARS-CoV-2, either in the laboratory or more importantly, in a field setting.

A novel electrochemical biosensor for DNA detection based on exonuclease III-assisted target recycling and rolling circle amplification

RSC Advances ◽  
2015 ◽  
Vol 5 (12) ◽  
pp. 9123-9129 ◽  
Author(s):  
Fenglei Gao ◽  
Yan Du ◽  
Jingwen Yao ◽  
Yanzhuo Zhang ◽  
Jian Gao
Keyword(s):  
Electrochemical Detection ◽  
Electrochemical Biosensor ◽  
Dna Detection ◽  
Rolling Circle Amplification ◽  
Exonuclease Iii ◽  
Target Recycling ◽  
Rolling Circle

A strategy for electrochemical detection of DNA by exonuclease III-assisted DNA recycling and the rolling circle amplification was developed.

Ultrasensitive electrochemical detection of breast cancer cells based on DNA-rolling-circle-amplification-directed enzyme-catalyzed polymerization

2015 ◽  
Vol 51 (11) ◽  
pp. 2114-2117 ◽  
Author(s):  
Qinglin Sheng ◽  
Ni Cheng ◽  
Wushuang Bai ◽  
Jianbin Zheng
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Electrochemical Detection ◽  
Breast Cancer Cells ◽  
Rolling Circle Amplification ◽  
Rolling Circle ◽  
Catalyzed Polymerization

An ultrasensitive cytosensor based on DNA-rolling-circle-amplification-directed enzyme-catalyzed polymerization is demonstrated.

scholarly journals On the Electrochemical Detection of Alpha-Fetoprotein Using Aptamers: DNA Isothermal Amplification Strategies to Improve the Performance of Weak Aptamers

Biosensors ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 46
Author(s):  
Ramón Lorenzo-Gómez ◽  
Daniel González-Robles ◽  
Rebeca Miranda-Castro ◽  
Noemí de-los-Santos-Álvarez ◽  
María Jesús Lobo-Castañón
Keyword(s):  
Dissociation Constant ◽  
Electrochemical Detection ◽  
Magnetic Beads ◽  
Rolling Circle Amplification ◽  
Binding Constants ◽  
Alpha Fetoprotein ◽  
Isothermal Amplification ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tumor Biomarker ◽  
Rolling Circle

Affinity characterization is essential to develop reliable aptamers for tumor biomarker detection. For alpha-fetoprotein (AFP), a biomarker of hepatocellular carcinoma (HCC), two DNA aptamers were described with very different affinity. In this work, we estimate the dissociation constant of both of them by means of a direct assay on magnetic beads modified with AFP and electrochemical detection on carbon screen-printed electrodes (SPCE). Unlike previous works, both aptamers showed similar dissociation constant (Kd) values, in the subµM range. In order to improve the performance of these aptamers, we proposed the isothermal amplification of the aptamers by both terminal deoxynucleotidyl transferase (TdT) and rolling circle amplification (RCA). Both DNA amplifications improved the sensitivity and also the apparent binding constants from 713 nM to 189 nM for the short aptamer and from 526 nM to 32 nM for the long aptamer. This improvement depends on the true affinity of the binding pair, which ultimately limits the analytical usefulness.

Sign in / Sign up

Export Citation Format

Share Document