Ultrasensitive DNA Microarray Biosensing via in Situ RNA Transcription-Based Amplification and Nanoparticle-Enhanced SPR Imaging
- 10 March 2011
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 133 (12), 4271-4273
- https://doi.org/10.1021/ja2005576
Abstract
DNA microarrays are invaluable tools for the detection and identification of nucleic acids in biosensing applications. The sensitivity and selectivity of multiplexed single-stranded DNA (ssDNA) surface bioaffinity sensing can be greatly enhanced when coupled to a surface enzymatic reaction. Herein we describe a novel method where the specific sequence-dependent adsorption of a target ssDNA template molecule onto an ssDNA-modified gold microarray is followed with the generation of multiple copies of ssRNA via in situ surface transcription by RNA polymerase. The RNA created on this “generator” element is then detected by specific adsorption onto a second adjacent “detector” element of ssDNA that is complementary to one end of the ssRNA transcript. SPR imaging is then used to detect the subsequent hybridization of cDNA-coated gold nanoparticles with the surface-bound RNA. This RNA transcription-based, dual element amplification method is used to detect ssDNA down to a concentration of 1 fM in a volume of 25 μL (25 zeptomoles).This publication has 18 references indexed in Scilit:
- Attomole Detection of Mesophilic DNA Polymerase Products by Nanoparticle-Enhanced Surface Plasmon Resonance Imaging on Glassified Gold SurfacesJournal of the American Chemical Society, 2010
- Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrationsNature Biotechnology, 2010
- Fabrication of Silica-Coated Gold Nanorods Functionalized with DNA for Enhanced Surface Plasmon Resonance Imaging Biosensing ApplicationsLangmuir, 2009
- Fabrication of DNA Microarrays with Poly(l-glutamic acid) Monolayers on Gold Substrates for SPR Imaging MeasurementsLangmuir, 2009
- Nanoparticle diffraction gratings for DNA detection on photopatterned glass substratesBiointerphases, 2008
- Microarray methods for protein biomarker detectionThe Analyst, 2008
- Multiplexed Detection Methods for Profiling MicroRNA Expression in Biological SamplesAngewandte Chemie International Edition, 2008
- Attomole Microarray Detection of MicroRNAs by Nanoparticle-Amplified SPR Imaging Measurements of Surface Polyadenylation ReactionsJournal of the American Chemical Society, 2006
- Creating Advanced Multifunctional Biosensors with Surface Enzymatic TransformationsLangmuir, 2006
- Single-Nucleotide Polymorphism Genotyping by Nanoparticle-Enhanced Surface Plasmon Resonance Imaging Measurements of Surface Ligation ReactionsAnalytical Chemistry, 2006