Isothermal cycling and cascade signal amplification strategy for ultrasensitive colorimetric detection of nucleic acids
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- 作者：Mingdi Xu ; Yu He ; Zhuangqiang Gao ; Guonan Chen ; Dianping Tang
- 关键词：Nucleic acid ; Colorimetric assay ; Isothermal cycling signal amplification
- 刊名：Microchimica Acta
- 出版年：2015
- 出版时间：January 2015
- 年：2015
- 卷：182
- 期：1-2
- 页码：449-454
- 全文大小：1,002 KB
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- 刊物主题：Chemistry
Analytical Chemistry
Inorganic Chemistry
Physical Chemistry
Characterization and Evaluation Materials
Monitoring, Environmental Analysis and Environmental Ecotoxicology- 出版者：Springer Wien
- ISSN：1436-5073
文摘
We have designed a novel isothermal cascade signal-amplification strategy for ultrasensitive colorimetric determination of nucleic acids. It is based on double-cycling amplification with formation of DNAzyme via a polymerase-induced strand-displacement reaction and nicking endonuclease-assisted recycling. The assay makes use of a hairpin DNA, a short primer, KF-polymerase, and nicking endonuclease. The presence of a target DNA triggers the strand-displacement and polymerization reaction with the formation of numerous DNAzyme molecules. Upon addition of H2O2 to the resulting mixture, the H2O2 reacts with 2,2-azino-bis (3-ethylbenzothiozoline)-6-sulfonate to form a colored product in the aid of DNAzyme, which is quantified by photometry at 415?nm. Under optimal conditions, the assay allows target DNA to be determined at concentration as low as 0.6?aM. Graphical Abstract Isothermal cycling and cascade signal amplification strategy has been developed for ultrasensitive colorimetric determination of nucleic acids by using DNA polymerase-induced strand-displacement and nicking endonuclease-assisted recycling.