Issue 3, 2012

Signal amplification by rolling circle amplification on universal flaps yielded from target-specific invasive reaction

Abstract

Detection of nucleic acids with signal amplification is preferable in clinical diagnosis. A novel approach was developed for signal amplification by coupling invasive reaction with hyperbranched rolling circle amplification (HRCA). Invasive reaction, which does not rely on specific recognition sequences in a target but a specific structure formed by the specific binding of an upstream probe and a downstream probe to a target DNA, can generate thousands of flaps from one target DNA; then the flaps are ligated with padlock probes to form circles, which are the templates of HRCA. As HRCA amplicon sequence is free of target DNA sequence, signal amplification is achieved. Because flap sequence is the same to any target of interest, HRCA is universal; the detection cost is hence greatly reduced. The sensitivity of the proposed method is less than 1 fM artificial DNA targets; and the specificity of the method is high enough to discriminate one base difference in the target sequence. The feasibility was verified by detecting real biological samples from HBV carriers, indicating that the method is highly sensitive, cost-effective, and has a low risk of cross-contamination from amplicons. These properties should give great potential in clinical diagnosis.

Graphical abstract: Signal amplification by rolling circle amplification on universal flaps yielded from target-specific invasive reaction

Supplementary files

Article information

Article type
Paper
Submitted
22 Sep 2011
Accepted
25 Oct 2011
First published
07 Dec 2011

Analyst, 2012,137, 729-734

Signal amplification by rolling circle amplification on universal flaps yielded from target-specific invasive reaction

B. Zou, Y. Ma, H. Wu and G. Zhou, Analyst, 2012, 137, 729 DOI: 10.1039/C1AN15886D

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements