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Photoelectrochemical biosensor for highly sensitive detection of microRNA based on duplex-specific nuclease-triggered signal amplification

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Abstract

A selective and sensitive photoelectrochemical assay was proposed for microRNA detection based on the isothermal cycle hybridization amplification of microRNA-21 that originated from the specific cleavage activity of duplex-specific nuclease (DSN) toward a DNA probe in DNA-RNA double helix and in situ enzymatic production of electron donor of ascorbic acid. After the biotin-functionalized DNA probe hybridized with complementary target microRNA-21, the hybridized DNA probe could be cleaved by DSN and microRNA-21 was released back to the incubation solution. The released microRNA could be further hybridized with the remaining single-strand DNA probe; thus, the isothermal cycle hybridization amplification would be formed. As a result, little avidin-alkaline phosphatase conjugate could be captured due to the release of biotin originating from the DSN cleavage. The distinct photocurrent change between a control biosensor and the DSN cleavage biosensor achieved label-free microRNA-21 detection with the linear range from 1 to 500 fM. The fabricated biosensor showed high detection selectivity even for one-base mismatched sequence. The attempt was carried out to directly assay the expression level change of microRNA-21 in total RNA extracted from chicken fibroblast cells infected with subgroup J avian leukosis virus.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Nos. 21105056, 21375079), the Foundation of State Key Laboratory of Crop Biology (No. 2014KF12), and the Project of Development of Science and Technology of Shandong Province, China (No. 2013GZX20109).

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Authors and Affiliations

  1. College of Chemistry and Material Science, Shandong Agricultural University, Tai’an, 271018, Shandong, People’s Republic of China

    Bingchen Li, Xue Li, Mo Wang, Zhiqing Yang, Huanshun Yin & Shiyun Ai

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  1. Bingchen Li
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  2. Xue Li
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  3. Mo Wang
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  4. Zhiqing Yang
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  5. Huanshun Yin
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  6. Shiyun Ai
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Correspondence to Huanshun Yin or Shiyun Ai.

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Li, B., Li, X., Wang, M. et al. Photoelectrochemical biosensor for highly sensitive detection of microRNA based on duplex-specific nuclease-triggered signal amplification. J Solid State Electrochem 19, 1301–1309 (2015). https://doi.org/10.1007/s10008-015-2747-5

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  • DOI: https://doi.org/10.1007/s10008-015-2747-5

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