Hybridization chain reaction-based fluorescence immunoassay using DNA intercalating dye for signal readout

doi:10.1039/c4an00190g

	Hybridization chain reaction-based fluorescence immunoassay using DNA intercalating dye for signal readout
	Zhou, Ying-Lin; Deng, Yan; Nie, Ji; Zhang, Xiao-hui; Zhao, Ming-Zhe; Zhang, Xin-Xiang
刊名	ANALYST
	2014
卷号	139 期号:13 页码:3378-3383
ISSN号	0003-2654
DOI	10.1039/c4an00190g
文献子类	Article
英文摘要	A novel format of fluorescence immunosorbent assay based on the hybridization chain reaction (HCR) using a DNA intercalating dye for signal readout was constructed for the sensitive detection of targets, both in competitive and sandwich modes. In this platform, the capture and recognition processes are based on immunoreactions and the signal amplification depends on the enzyme-free, isothermal HCR-induced labelling event. After a competitive or a sandwich immunoreaction, a biotinylated capture DNA was bound to a biotinylated signal antibody through avidin, and triggered the HCR by two specific hairpins into a nicked double helix. Gene Finder (GF), a fluorescent probe for double-strand DNA, was intercalated in situ into the amplified chain to produce the fluorescence signal. The limit of detection (LOD) for rabbit IgG in competitive mode by HCR/GF immunoassay was improved at least 100-fold compared with the traditional fluorescence immunoassay using the fluorescein isothiocyanate-labelledstreptavidin or fluorescein isothiocyanate-labelled second antibody as the signal readout. The proposed fluorescence immunoassay was also demonstrated by using alpha-fetoprotein as the model target in sandwich mode, and showed a wide linear range from 28 ng mL(-1) to 20 mu g mL(-1) with a LOD of 6.0 ng mL(-1). This method also showed satisfactory analysis in spiked human serum, which suggested that it might have great potential for versatile applications in life science and point-of-care diagnostics.
WOS关键词	LINKED-IMMUNOSORBENT-ASSAY ; ANTIGEN-DETECTION ; AMPLIFICATION ; ENZYME ; CONJUGATE ; SAMPLES
WOS研究方向	Chemistry
语种	英语
出版者	ROYAL SOC CHEMISTRY
WOS记录号	WOS:000337125600025
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/167354]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Zhou, Ying-Lin
作者单位	Peking Univ, Beijing Natl Lab Mol Sci, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Coll Chem, Beijing 100871, Peoples R China
推荐引用方式 GB/T 7714	Zhou, Ying-Lin,Deng, Yan,Nie, Ji,et al. Hybridization chain reaction-based fluorescence immunoassay using DNA intercalating dye for signal readout[J]. ANALYST,2014,139(13):3378-3383.
APA	Zhou, Ying-Lin,Deng, Yan,Nie, Ji,Zhang, Xiao-hui,Zhao, Ming-Zhe,&Zhang, Xin-Xiang.(2014).Hybridization chain reaction-based fluorescence immunoassay using DNA intercalating dye for signal readout.ANALYST,139(13),3378-3383.
MLA	Zhou, Ying-Lin,et al."Hybridization chain reaction-based fluorescence immunoassay using DNA intercalating dye for signal readout".ANALYST 139.13(2014):3378-3383.