A colorimetric method for H1N1 DNA detection using rolling circle amplification

被引：41

作者：

Xing, Yasi ^{[1
,2
]}

Wang, Ping ^{[1
]}

Zang, Yucui ^{[1
,3
]}

Ge, Yuqing ^{[1
]}

Jin, Qinghui ^{[1
]}

Zhao, Jianlong ^{[1
]}

Xu, Xia ^{[2
]}

Zhao, Guoqiang ^{[3
]}

Mao, Hongju ^{[1
]}

机构：

[1] Chinese Acad Sci, Shanghai Inst Microsyst & Informat Technol, State Key Lab Transducer Technol, Shanghai 200050, Peoples R China

[2] Zhengzhou Univ, Coll Pharm, Zhengzhou 450001, Peoples R China

[3] Zhengzhou Univ, Dept Basic Med Coll, Zhengzhou 450052, Peoples R China

来源：

ANALYST | 2013年 / 138卷 / 12期

关键词：

POLYMERASE-CHAIN-REACTION; GOLD NANOPARTICLE PROBES; NUCLEIC-ACIDS; GENOMIC DNA; IDENTIFICATION; SEQUENCES; VIRUS;

D O I：

10.1039/c3an00100h

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A highly sensitive and specific colorimetry-based rolling circle amplification (RCA) assay has been successfully developed as a method for the effective detection of H1N1 DNA. Specific oligonucleotide and reporter primer probes were designed together with a circular template, and the oligonucleotide probes were attached to the surfaces of magnetic beads (MBs) to form functional MB-DNA conjugates as capture probes for the target H1N1 DNA molecules. Together with the addition of DNA targets and reporter primer probes to the MB-DNA conjugates, sandwiched hybrids were formed. The initiation of RCA amplification using the circular template in the presence of phi29 polymerase allowed for the amplification of a large number of repeat sequences of the single-stranded (ss)-DNA product. This RCA product accumulated gold nanoparticles (AuNPs), resulting in a colorimetric change that could be viewed by the naked eye or detected using UV-vis spectroscopy. According to this method, H1N1 DNA could be detected at the 1 pmol L-1 level. This platform exhibited design convenience, simplicity, and cost-effectiveness, and could be used to provide a new diagnostic assay for H1N1, and other infectious diseases.

引用

页码：3457 / 3462

页数：6

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