Nonenzymatic detection of bacterial genomic DNA using the bio bar code assay

被引:113
作者
Hill, Haley D. [1 ]
Vega, Rafael A. [1 ]
Mirkin, Chad A. [1 ]
机构
[1] Northwestern Univ, Int Inst Nanotechnol, Dept Chem, Evanston, IL 60208 USA
关键词
D O I
10.1021/ac701626y
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The detection of bacterial genomic DNA through a nonenzymatic nanomaterials-based amplification method, the bio bar code assay, is reported. The assay utilizes oligonucleotide-functionalized magnetic microparticles to capture the target of interest from the sample. A critical step in the new assay involves the use of blocking oligonucleotides during heat denaturation of the double-stranded DNA. These blockers bind to specific regions of the target DNA upon cooling and prevent the duplex DNA from rehybridizing, which allows the particle probes to bind. Following target isolation using the magnetic particles, oligonucleotide-functionalized gold nanoparticles act as target recognition agents. The oligonucleotides on the nanoparticle (bar codes) act as amplification surrogates. The bar codes are then detected using the Scanomettic method. The limit of detection for this assay was determined to be 2.5 fM, and this is the first demonstration of a bar code-type assay for the detection of double-stranded, genomic DNA.
引用
收藏
页码:9218 / 9223
页数:6
相关论文
共 43 条
[11]   Colorimetric screening of DNA-binding molecules with gold nanoparticle probes [J].
Han, MS ;
Lytton-Jean, AKR ;
Oh, BK ;
Heo, J ;
Mirkin, CA .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2006, 45 (11) :1807-1810
[12]   Colloidal Au-enhanced surface plasmon resonance for ultrasensitive detection of DNA hybridization [J].
He, L ;
Musick, MD ;
Nicewarner, SR ;
Salinas, FG ;
Benkovic, SJ ;
Natan, MJ ;
Keating, CD .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2000, 122 (38) :9071-9077
[13]   Real time quantitative PCR [J].
Heid, CA ;
Stevens, J ;
Livak, KJ ;
Williams, PM .
GENOME RESEARCH, 1996, 6 (10) :986-994
[14]   KINETIC PCR ANALYSIS - REAL-TIME MONITORING OF DNA AMPLIFICATION REACTIONS [J].
HIGUCHI, R ;
FOCKLER, C ;
DOLLINGER, G ;
WATSON, R .
BIO-TECHNOLOGY, 1993, 11 (09) :1026-1030
[15]   The bio-barcode assay for the detection of protein and nucleic acid targets using DTT-induced ligand exchange [J].
Hill, Haley D. ;
Mirkin, Chad A. .
NATURE PROTOCOLS, 2006, 1 (01) :324-336
[16]   Aptamer-modified gold nanoparticles for colorimetric determination of platelet-derived growth factors and their receptors [J].
Huang, CC ;
Huang, YF ;
Cao, ZH ;
Tan, WH ;
Chang, HT .
ANALYTICAL CHEMISTRY, 2005, 77 (17) :5735-5741
[17]   Maximizing DNA loading on a range of gold nanoparticle sizes [J].
Hurst, Sarah J. ;
Lytton-Jean, Abigail K. R. ;
Mirkin, Chad A. .
ANALYTICAL CHEMISTRY, 2006, 78 (24) :8313-8318
[18]   What controls the melting properties of DNA-linked gold nanoparticle assemblies? [J].
Jin, RC ;
Wu, GS ;
Li, Z ;
Mirkin, CA ;
Schatz, GC .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (06) :1643-1654
[19]  
JOCHENWILHELM AP, 2003, CHEMBIOCHEM, V4, P1120
[20]  
KARY BM, 1994, ANGEW CHEM INT EDIT, V33, P1209