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Multiplexed fluorometric determination for three microRNAs in acute myocardial infarction by using duplex-specific nuclease and MoS2 nanosheets

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Abstract

Circulating microRNAs are of diagnostic value for acute myocardial infarction (AMI). This study describes a fluorometric assay for multiplexed detection of the AMI biomarkers microRNA-499, microRNA-133a and microRNA-1. The assay involves the following two steps: (a) duplex-specific nuclease (DSN)-mediated signal amplification using aptamer-based sensor; (b) MoS2 nanosheets-based multiplexed fluorometric signal detection, and fluorometric signals have excitation/emission maxima at 492/518 nm for microRNA-499, 565/580 nm for microRNA-133a and 649/663 nm for microRNA-1. The assay has detection limits of around 100 fM for all three microRNAs. The assay is highly specific and rapid. It demonstrates that the expressions of microRNA-499, microRNA-133a and microRNA-1 are significantly higher in AMI patients. The ROC curves allow a clear distinction to be made between AMI group and non-AMI group.

Schematic representation of a multiplexed fluorometric method based on duplex-specific nuclease (DSN)-mediated signal amplification and MoS2 nanosheets-based fluorometric signal detection (DSN-MoS2) for rapid, sensitive and specific detection of microRNAs in AMI.

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Author contribution statement

Chengjian Yang and Zhijun Han design experiments; Yan Jin, Shuya Wang, Xiaoxiao Liu and Haohao Liu carry out experiments; Xue Zhu, Ke Wang and Peiling Zhou analyze experimental results; Xue Zhu and Ke Wang write the manuscript.

Funding

This work was supported by grant from Natural Science Foundation of Jiangsu Province (No. BK20171147), Jiangsu Young Medical Talents (QNRC2016167), Six talent peaks project in Jiangsu Province (WSW-047), The Science and Technology Projects of Wuxi City (WX18IVJN016) and The Major Science and Technology Projects of Wuxi City (Z201804, MS201653).

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  1. Xue Zhu and Ke Wang contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063, Jiangsu Province, People’s Republic of China

    Xue Zhu & Ke Wang

  2. Department of Cardiology, The Affiliated Wuxi NO.2 People’s Hospital of Nanjing Medical University, Wuxi, 214002, Jiangsu Province, People’s Republic of China

    Yan Jin, Shuya Wang, Xiaoxiao Liu, Peiling Zhou & Chengjian Yang

  3. Department of Laboratory Medicine, The Affiliated Wuxi NO.2 People’s Hospital of Nanjing Medical University, Wuxi, 214002, Jiangsu Province, People’s Republic of China

    Haohao Liu & Zhijun Han

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  1. Xue Zhu
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Zhu, X., Wang, K., Jin, Y. et al. Multiplexed fluorometric determination for three microRNAs in acute myocardial infarction by using duplex-specific nuclease and MoS2 nanosheets. Microchim Acta 187, 15 (2020). https://doi.org/10.1007/s00604-019-3896-5

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