Ultrasensitive Detection of DNA and RNA Based on Enzyme-Free Click Chemical Ligation Chain Reaction on Dispersed Gold Nanoparticles

被引:66
作者
Kato, Daiki [1 ]
Oishi, Motoi [1 ]
机构
[1] Univ Tsukuba, Fac Pure & Appl Sci, Div Mat Sci, Tsukuba, Ibaraki 3058573, Japan
基金
日本学术振兴会;
关键词
click chemistry; DNA; gold nanoparticles; ligation chain reaction; magnetic beads; RNA; COLORIMETRIC DETECTION; POLYNUCLEOTIDE LIGASE; OPTICAL-PROPERTIES; COPPER-FREE; OLIGONUCLEOTIDES; AMPLIFICATION; CHEMISTRY; BIOLOGY; ACID;
D O I
10.1021/nn503150w
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
An ultrasensitive colorimetric DNA and RNA assay using a combination of enzyme-free click chemical ligation chain reaction (CCLCR) on dispersed gold nanoparticles (GNPs) and a magnetic separation process has been developed. The click chemical ligation between an azide-containing probe DNA-modified GNP and a dibenzocyclooctyne-containing probe biotinyl DNA occurred through hybridization with target DNA (RNA) to form the biotinyl-ligated GNPs (ligated products). Eventually, both the biotinyl-ligated GNPs and target DNA (RNA) were amplified exponentially using thermal cycling. After separation of the biotinyl-ligated GNPs using streptavidin-modified magnetic beads, the change in intensity of the surface plasmon band at 525 nm in the supernatants was observed by UV/vis measurement and was also evident visually. The CCLCR assay provides ultrasensitive detection (50 zM: several copies) of target DNA that is comparable to PCR-based approaches. Note that target RNA could also be detected with similar sensitivity without the need for reverse transcription to the corresponding cDNA. The amplification efficiency of the CCLCR assay was as high as 82% due to the pseudohomogeneous reaction behavior of CCLCR on dispersed GNPs. In addition, the CCLCR assay was able to discriminate differences in single-base mismatches and to specifically detect target DNA and target RNA from the cell lysate.
引用
收藏
页码:9988 / 9997
页数:10
相关论文
共 37 条
[31]   What controls the optical properties of DNA-linked gold nanoparticle assemblies? [J].
Storhoff, JJ ;
Lazarides, AA ;
Mucic, RC ;
Mirkin, CA ;
Letsinger, RL ;
Schatz, GC .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2000, 122 (19) :4640-4650
[32]   One-pot colorimetric differentiation of polynucleotides with single base imperfections using gold nanoparticle probes [J].
Storhoff, JJ ;
Elghanian, R ;
Mucic, RC ;
Mirkin, CA ;
Letsinger, RL .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1998, 120 (09) :1959-1964
[33]   p24 antigen detection assay modified with a booster step for diagnosis and monitoring of human immunodeficiency virus type 1 infection [J].
Sutthent, R ;
Gaudart, N ;
Chokpaibulkit, K ;
Tanliang, N ;
Kanoksinsombath, C ;
Chaisilwatana, P .
JOURNAL OF CLINICAL MICROBIOLOGY, 2003, 41 (03) :1016-1022
[34]   Wnt Signalling Pathway Parameters for Mammalian Cells [J].
Tan, Chin Wee ;
Gardiner, Bruce S. ;
Hirokawa, Yumiko ;
Layton, Meredith J. ;
Smith, David W. ;
Burgess, Antony W. .
PLOS ONE, 2012, 7 (02)
[35]   Development of an aggregation-based immunoassay for anti-protein A using gold nanoparticles [J].
Thanh, NTK ;
Rosenzweig, Z .
ANALYTICAL CHEMISTRY, 2002, 74 (07) :1624-1628
[36]   Gold-Nanoparticle-Based Colorimetric Discrimination of Cancer-Related Point Mutations with Picomolar Sensitivity [J].
Valentini, Paola ;
Fiammengo, Roberto ;
Sabella, Stefania ;
Gariboldi, Manuela ;
Maiorano, Gabriele ;
Cingolani, Roberto ;
Pompa, Pier Paolo .
ACS NANO, 2013, 7 (06) :5530-5538
[37]   Ligation Chain Reaction based gold nanoparticle assembly for ultrasensitive DNA detection [J].
Yin, Honghong ;
Huang, Xin ;
Ma, Wei ;
Xu, Liguang ;
Zhu, Shuifang ;
Kuang, Hua ;
Xu, Chuanlai .
BIOSENSORS & BIOELECTRONICS, 2014, 52 :8-12