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Isothermal cycling and cascade signal amplification strategy for ultrasensitive colorimetric detection of nucleic acids

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Abstract

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.

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.

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Acknowledgments

Support from the “973” National Basic Research Program of China (2010CB732403), the NSFC of China (41176079), the NSF of Fujian Province (2011J06003), and the Program for Changjiang Scholars and Innovative Research Team in University (IRT1116) is gratefully acknowledged.

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Authors and Affiliations

  1. Institute of Nanomedicine and Nanobiosensing, Department of Chemistry, Fuzhou University, Fuzhou 350108, People’s Republic of China

    Mingdi Xu, Yu He, Zhuangqiang Gao, Guonan Chen & Dianping Tang

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  1. Mingdi Xu
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  2. Yu He
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  3. Zhuangqiang Gao
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  4. Guonan Chen
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  5. Dianping Tang
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Correspondence to Yu He or Dianping Tang.

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Xu, M., He, Y., Gao, Z. et al. Isothermal cycling and cascade signal amplification strategy for ultrasensitive colorimetric detection of nucleic acids. Microchim Acta 182, 449–454 (2015). https://doi.org/10.1007/s00604-014-1385-4

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  • DOI: https://doi.org/10.1007/s00604-014-1385-4

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