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A polyaniline-reduced graphene oxide nanocomposite as a redox nanoprobe in a voltammetric DNA biosensor for Mycobacterium tuberculosis

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Abstract

This article describes the construction of a sandwich-type of DNA biosensor for detecting a specific DNA sequence of Mycobacterium tuberculosis (MTB). A polyaniline-reduced graphene oxide (PANI-rGO) composite with favorable electrochemical activity was used as a redox nanoprobe for the generation of the voltammetric signal. The composite was decorated with gold nanoparticles (AuNPs) onto which the signal probe was immobilized to form the tracer label. After hybridization between target DNA and tracer label, the voltammetric signal resulting from the polyaniline-reduced graphene oxide (PANI-rGO) redox probe can be apparently observed. The biosensor can detect the specific IS6110 DNA sequence of MTB in the 0.1 pM to 10 nM concentration range, and the detection limit is as low as 50 fM (at an S/N ratio of 3). The biosensor is also highly specific and does not recognize mismatches. It was applied to the determination of denatured PCR products in clinical samples such as sputum, and the results agreed with those obtained by gel electrophoresis. This assay provides a versatile and powerful tool for detection of MTB, and probably for other pathogens if appropriate molecular markers are available.

Schematic of an electrochemical DNA biosensor for the ultrasensitive detection of Mycobacterium tuberculosis (MTB). It uses a redox nanoprobe (rGO-PANI) for signal amplification. MCH: 6-mercapto-1-hexanol; PANI: polyaniline; rGO: reduced graphene oxide; rGO-PANI: polyaniline-reduced graphene oxide nanocomposite.

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Acknowledgements

This work is supported by National Natural Science Foundation of China (81601856), Funds for Young Science and Technology Talent Cultivation Plan of Chongqing City (cstc2014kjrc-qnrc00004), the National Key Clinical Specialist Construction Programs of China ([2012] No. 649) and Funds for Outstanding Young Scholars in Chongqing Medical University (CYYQ201405).

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  1. Yuhan Chen and Youlun Li contributed equally to this work

Authors and Affiliations

  1. Department of Respiratory Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Yuhan Chen, Youlun Li, Yang Yang, Fangni Wu & Jun Cao

  2. Pharmaceutical Engineering Research Center, College of Pharmacy, Chongqing Medical University, No.1 Yixueyuan Road, Chongqing, 400016, China

    Lijuan Bai

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  1. Yuhan Chen
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  2. Youlun Li
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  3. Yang Yang
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Correspondence to Lijuan Bai.

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No conflict of interest exits in the submission of this manuscript, and it is approved by all authors for publication. All procedures performed in this study were in accordance with the ethical standards.

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Chen, Y., Li, Y., Yang, Y. et al. A polyaniline-reduced graphene oxide nanocomposite as a redox nanoprobe in a voltammetric DNA biosensor for Mycobacterium tuberculosis . Microchim Acta 184, 1801–1808 (2017). https://doi.org/10.1007/s00604-017-2184-5

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  • DOI: https://doi.org/10.1007/s00604-017-2184-5

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