Molecular beacon structure mediated rolling circle amplification for ultrasensitive electrochemical detection of microRNA based on quantum dots tagging

2013 ◽  
Vol 33 ◽  
pp. 80-83 ◽  
Author(s):  
Danchen Wang ◽  
Lihui Hu ◽  
Huiming Zhou ◽  
E.S. Abdel-Halim ◽  
Jun-Jie Zhu
Keyword(s):  
Quantum Dots ◽  
Electrochemical Detection ◽  
Molecular Beacon ◽  
Rolling Circle Amplification ◽  
Rolling Circle

Molecular Beacon Enables Combination of Highly Processive and Highly Sensitive Rolling Circle Amplification Readouts for Detection of DNA-Modifying Enzymes

Nano LIFE ◽  
2015 ◽  
Vol 05 (02) ◽  
pp. 1541002 ◽  
Author(s):  
Emil L. Kristoffersen ◽  
Maria Gonzalez ◽  
Magnus Stougaard ◽  
Cinzia Tesauro
Keyword(s):  
Real Time ◽  
Dna Cleavage ◽  
Topoisomerase I ◽  
Drug Response ◽  
Molecular Beacon ◽  
Rolling Circle Amplification ◽  
Dna Topoisomerase ◽  
Rolling Circle ◽  
Topoisomerase Ib ◽  
Highly Sensitive

Here we present an optimized readout format for detection of the circularized products from a DNA-based sensor for measurement of DNA-modifying enzymes including DNA Topoisomerase I. The basic design of the DNA-sensor relies on the use of a substrate that can be circularized by the activity of DNA-modifying enzymes like type IB Topoisomerases and subsequently amplified by a rolling circle amplification (RCA) mechanism. The RCA process can be followed in real-time by the addition of a molecular beacon with a fluorophore/quencher pair. Upon hybridization to the amplified product, the fluorophore/quencher pair is separated, giving rise to a fluorescent signal, measurable in pseudo real-time using a qPCR machine or in a fluorimeter. The RCA products in complex with the molecular beacon can subsequently be moved to microscopic slides and analyzed in a fluorescence microscope. We describe the proof of the principle of this molecular beacon-based method combining the qPCR readout format with the standard Rolling circle Enhanced Enzymatic Assay previously reported. Although the qPCR setup is less sensitive, it allows easy, fast, and high-throughput measurement of enzyme activities. Human Topoisomerase IB (TopIB) is a well-known target for the clinically used anticancer drugs of the camptothecin family. The cytotoxic effect of camptothecins correlates directly with the intracellular TopIB activity affecting reversibly the Topoisomerase/DNA cleavage complexes. Therefore, we envisioned that the presented method may find use for the prediction of cellular drug response and for drug screening to discover novel molecules that specifically inhibit TopIB or other DNA-modifying enzymes.

A linear DNA probe as an alternative to a molecular beacon for improving the sensitivity of a homogenous fluorescence biosensing platform for DNA detection using target-primed rolling circle amplification

RSC Advances ◽  
2015 ◽  
Vol 5 (6) ◽  
pp. 4019-4025 ◽  
Author(s):  
Fulin Zhou ◽  
Baoxin Li ◽  
Jiyuan Ma
Keyword(s):  
Fluorescence Quenching ◽  
Molecular Beacon ◽  
Dna Detection ◽  
Rolling Circle Amplification ◽  
Dna Probes ◽  
Dna Probe ◽  
Rolling Circle ◽  
Hairpin Probe ◽  
Linear Dna ◽  
Signal Probe

Linear single-labeled DNA probes are used in this RCA-based fluorescence strategy for DNA detection, which could effectively avoid the fluorescence quenching between neighboring signal probes using hairpin probe as signal probe.

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

Highly Sensitive MicroRNA Detection by Coupling Nicking-Enhanced Rolling Circle Amplification with MoS2 Quantum Dots

2020 ◽  
Vol 92 (19) ◽  
pp. 13588-13594
Author(s):  
Jia Ge ◽  
Yun Hu ◽  
Ruijie Deng ◽  
Zhaohui Li ◽  
Kaixiang Zhang ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Rolling Circle Amplification ◽  
Rolling Circle ◽  
Microrna Detection ◽  
Highly Sensitive

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.

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