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Aptamer-mediated colorimetric and electrochemical detection of Pseudomonas aeruginosa utilizing peroxidase-mimic activity of gold NanoZyme

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Abstract

Despite of various advancements in biosensing, a rapid, accurate, and on-site detection of a bacterial pathogen is a real challenge due to the lack of appropriate diagnostic platforms. To address this unmet need, we herein report an aptamer-mediated tunable NanoZyme sensor for the detection of Pseudomonas aeruginosa, an infectious bacterial pathogen. Our approach exploits the inherent peroxidase-like NanoZyme activity of gold nanoparticles (GNPs) in combination with high affinity and specificity of a Pseudomonas aeruginosa–specific aptamer (F23). The presence of aptamer inhibits the inherent peroxidase-like activity of GNPs by simple adsorption on to the surface of GNPs. However, in the presence of cognate target (P. aeruginosa), owing to the high affinity for P. aeruginosa, the aptamer leaves the GNP surface, allowing GNPs to resume their peroxidase-like activity, resulting in oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB). As TMB is an electrochemically active species, we have been able to translate the NanoZyme-based method into an ultrasensitive electrochemical assay using disposable carbon screen-printed electrode. This approach is highly sensitive and allows us to rapidly detect P. aeruginosa with a low-end detection limit of ~ 60 CFU/mL in water within 10 min. This generic aptamer-NanoZyme-based electrochemical sensing strategy may, in principle, be applicable for the detection of various other bacterial pathogens.

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Acknowledgements

The authors acknowledge the generous support provided by Prof. Jaya S. Tyagi and Dr. H.K. Prasad in providing laboratory access to handle the bacterial cultures.

Funding

T.K.S. is thankful to THSTI Core grant and the Department of Biotechnology Govt. of India for Innovation Award and Innovative Young Biotechnologist Award (BT/010/IYBA/2016/10). V.B. thanks the Australian Research Council (ARC) for a Future Fellowship (FT140101285) and research support through an ARC Discovery (DP170103477) grant. V.B. also recognizes the generous support of the Ian Potter Foundation toward establishing the Sir Ian Potter NanoBioSensing Facility at RMIT University. A.D. is thankful to the Indian Council for Medical Research for providing Senior Research Fellowship.

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Author notes

  1. Ritu Das

    Present address: Indian Institute of Technology-Delhi (IIT-Delhi), New Delhi, 110016, India

  2. Ritu Das and Abhijeet Dhiman contributed equally to this work.

Authors and Affiliations

  1. AptaBharat Innovation Pvt. Ltd., Interim Incubator of Translational Health Science and Technology Institute (THSTI), Faridabad, Haryana, 121001, India

    Ritu Das & Tarun Kumar Sharma

  2. Department of Biotechnology, All India Institute of Medical Sciences (AIIMS), New Delhi, 110029, India

    Abhijeet Dhiman

  3. Faculty of Pharmacy, Uttarakhand Technical University (UTU), Dehradun, Uttarakhand, 248007, India

    Abhijeet Dhiman

  4. Department of Microbiology, All India Institute of Medical Sciences (AIIMS), New Delhi, 110029, India

    Arti Kapil

  5. Ian Potter NanoBioSensing Facility, NanoBiotechnology Research Lab (NBRL), School of Science, RMIT University, GPO Box 2476, Melbourne, VIC, 3001, Australia

    Vipul Bansal

  6. Centre for Biodesign and Diagnostics, Translational Health Science and Technology Institute (THSTI), Faridabad, Haryana, 121001, India

    Tarun Kumar Sharma

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  1. Ritu Das
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Correspondence to Vipul Bansal or Tarun Kumar Sharma.

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Das, R., Dhiman, A., Kapil, A. et al. Aptamer-mediated colorimetric and electrochemical detection of Pseudomonas aeruginosa utilizing peroxidase-mimic activity of gold NanoZyme. Anal Bioanal Chem 411, 1229–1238 (2019). https://doi.org/10.1007/s00216-018-1555-z

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