A Label-free Aptasensor for Rapid Detection of H1N1 Virus based on Graphene Oxide and Polymerase-aided Signal Amplification
- *Corresponding Author:
- Le Deng
Department of Microbiology
College of Life Science
Hunan Normal University
Changsha, Hunan, 410081, P.R China
Fax: 86-0731 88883310
E-mail: [email protected]
Received Date: March 09, 2015; Accepted Date: April 13, 2015; Published Date: May 05, 2015
Citation: Feng X, Liu K, Ning Y, Chen L, Deng L (2015) A Label-free Aptasensor for Rapid Detection of H1N1 Virus based on Graphene Oxide and Polymeraseaided Signal Amplification. J Nanomed Nanotechnol 6:288. doi:10.4172/2157-7439.1000288
Copyright: © 2015 Feng X, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Single-stranded DNA aptamers specific to the hemagglutinin (HA) protein of avian influenza virus (A/Puerto Rico/8/1934) were obtained by SELEX process in 13 cycles. The aptamer with the highest affinity and specificity was applied to an affinity bioassay. An aptamer hairpin (aHP) was prepared that consists of two DNA regions, viz. (a) the aptamer for the HA protein and (b) an oligonucleotide designed to form a stem-loop structure. In the absence of target, the aHP maintains its hairpin structure and was adsorbed to the Graphene Oxide (GO). The fluorescence of SYBR Green I (SGI) is almost quenched. On addition of the target, aHP unfolds and the GO is no longer attached but released to the solution. By applying a polymerase elongation reaction, a long dsDNA product is generated when SGI is added. The proposed method could detect HA and H1N1 virus with a limit of 2.5 μg/mL and 1×102 TCID50, respectively. Consequently, this paves the way for influenza virus detection and is employed in basic research and medical diagnosis.