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Dual-aptamer-based voltammetric biosensor for the Mycobacterium tuberculosis antigen MPT64 by using a gold electrode modified with a peroxidase loaded composite consisting of gold nanoparticles and a Zr(IV)/terephthalate metal-organic framework

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Abstract

An ultrasensitive aptasensor is described for the voltammetric determination of the Mycobacterium tuberculosis antigen MPT64 in human serum. Firstly, an amino-modified Zr(IV) based metal-organic framework (MOF; type UiO-66-NH2; made up from Zr6O32 units and 2-amino-terephthalate linkers) with a high specific surface was synthesized and used as the carrier of the gold nanoparticles and the aptamers. Then the signalling nanoprobe was fabricated after the horseradish peroxidase was cast on the nanomaterials. The two aptamers with synergistic effect on binding MPT64 were anchored on the gold electrode. Differential pulse voltammetry indicated that the peak current is highest if the ratio of the two aptamers is 1:1. The assay has a wide linear response range (0.02 to 1000 pg·mL−1 of MPT64) and a 10 fg·mL−1 detection limit at a working potential of around −96 mV (vs Ag/AgCl). The results show this biosensor to be a viable tool for detection of tuberculosis at an early stage.

Schematic presentation of the construction of the nanoprobe and biosensor. Firstly, the surface of UiO-66-NH2 was anchored to gold nanoparticles (AuNPs). A dual-aptamer and HRP were added to form the signalling nanoprobe (Aptamer/HRP/AuNPs/UiO-66-NH2). Then, the aptamers I and II were attached on the surface of gold electrode and 6-mercapto-1-hexanol was used to block the uncovered active site of the gold electrode. Finally, after incubation with MPT64, the signalling nanoprobe was dropped on the modified electrode and the DPV measurements was used for the analysis of Mycobacterium tuberculosis antigen MPT64. (PVP: poly(vinyl pyrrolidone); HRP: horseradish peroxidase; MCH: 6-Mercapto-1-hexanol; HQ: hydroquinone; BQ: benzoquinone).

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 21675177), the Science and Technology Planning Project of Guangdong Province (No. 2016B030303002), the Natural Science Foundation of Guangdong Province (No. 2018A030310142), and the Medical Scientific Research Foundation of Guangdong Province (No. A2017033).

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou, 510006, China

    Ningning Li, Xing Huang, Duanping Sun & Zuanguang Chen

  2. Center for Drug Research and Development, Guangdong Pharmaceutical University, Guangzhou, 510006, China

    Duanping Sun

  3. Shenzhen Center for Chronic Disease Control, Shenzhen, 518020, China

    Weiye Yu & Weiguo Tan

  4. The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, 518071, China

    Zhaofan Luo

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  1. Ningning Li
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  2. Xing Huang
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Correspondence to Duanping Sun or Zuanguang Chen.

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Li, N., Huang, X., Sun, D. et al. Dual-aptamer-based voltammetric biosensor for the Mycobacterium tuberculosis antigen MPT64 by using a gold electrode modified with a peroxidase loaded composite consisting of gold nanoparticles and a Zr(IV)/terephthalate metal-organic framework. Microchim Acta 185, 543 (2018). https://doi.org/10.1007/s00604-018-3081-2

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