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Molecular beacon based biosensor for the sequence-specific detection of DNA using DNA-capped gold nanoparticles-streptavidin conjugates for signal amplification

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Abstract

We describe a highly sensitive and selective molecular beacon-based electrochemical impedance biosensor for the sequence-specific detection of DNA. DNA-capped conjugates between gold nanoparticles (Au-NPs) and streptavidin are used for signal amplification. The molecular beacon was labeled with a thiol at its 5′ end and with biotin at its 3′ end, and then immobilized on the surface of a bare gold electrode through the formation of Au-S bonds. Initially, the molecular beacon is present in the “closed” state, and this shields the biotin from being approached by streptavidin due to steric hindrance. In the presence of the target DNA, the target DNA molecules hybridize with the loop and cause a conformational change that moves the biotin away from the surface of the electrode. The biotin thereby becomes accessible for the reporter (the DNA-streptavidin capped Au-NPs), and this results in a distinct increase in electron transfer resistance. Under optimal conditions, the increase in resistance is linearly related to the logarithm of the concentration of complementary target DNA in the range from 1.0 fM to 0.1 μM, with a detection limit of 0.35 fM (at an S/N of 3). This biosensor exhibits good selectivity, and acceptable stability and reproducibility.

We fabricated a novel sensitive electrochemical DNA biosensor based on the molecular beacon and conjugates composed of report DNA, Au-NPs and streptavidin (DAS) amplification signal protocol. The biosensor exhibits high sensitivity and good specificity even for single-mismatched DNA detection.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (No. 20675002).

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Authors and Affiliations

  1. College of Chemistry and Materials Science, Anhui Key Laboratory of Chemo-Biosensing, Anhui Normal University, Wuhu, 241000, People’s Republic of China

    Xian Fang, Wei Jiang, Xiaowei Han & Yuzhong Zhang

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  1. Xian Fang
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  2. Wei Jiang
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  3. Xiaowei Han
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  4. Yuzhong Zhang
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Correspondence to Yuzhong Zhang.

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Fang, X., Jiang, W., Han, X. et al. Molecular beacon based biosensor for the sequence-specific detection of DNA using DNA-capped gold nanoparticles-streptavidin conjugates for signal amplification. Microchim Acta 180, 1271–1277 (2013). https://doi.org/10.1007/s00604-013-1044-1

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  • DOI: https://doi.org/10.1007/s00604-013-1044-1

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