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Portable electrochemical immunosensor for detection of Mycobacterium tuberculosis secreted protein CFP10-ESAT6 in clinical sputum samples

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Abstract

An early detection of Mycobacterium tuberculosis is very important to reduce the number of fatal cases and allow for fast recovery. However, the interpretation of the result from smear microscopy requires skilled personnel due to the propensity of the method to produce false-negative results. In this work, a portable, rapid, and simple sandwich-type immunosensor reader has been developed that is able to detect the presence of M. tuberculosis in sputum samples. By using sandwich-type immunosensor, an anti-CFP10-ESAT6 antibody was immobilized onto the graphene/polyaniline (GP/PANI)-modified gold screen-printed electrode. After incubation with the target CFP10-ESAT6 antigen, the iron/gold magnetic nanoparticles (Fe3O4/Au MNPs) conjugated with anti-CFP10-ESAT6 antibody were used to complete the sandwich format. Differential pulse voltammetry (DPV) technique was used to detect the CFP10-ESAT6 antigen at the potential range of 0.0–1.0 V. The detection time is less than 2 h. Under optimal condition, CFP10-ESAT6 antigen was detected in a linear range from 10 to 500 ng mL−1 with a limit of detection at 1.5 ng mL−1. The method developed from this process was then integrated into a portable reader. The performance of the sensor was investigated and compared with the standard methods (culture and smear microscopy). It provides a good correlation (100% sensitivity and 91.7% specificity) with both methods of detection for M. tuberculosis in sputum samples henceforth, demonstrating the potential of the device as a more practical screening tool.

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Acknowledgments

The author would like to thank BP healthcare for providing the sputum samples.

Funding

This research was supported by NanoMalaysia Berhad, a CLG under the Ministry of Energy, Science, Technology, Environment and Climate Change (MESTECC), Malaysia (Project No. P17014), Malaysia Research University Network (MRUN) (UPM/800-4/11/MRUN/2018/5539230), and Putra Grant-Putra Graduate Initiative (GP-IPS) under Universiti Putra Malaysia (9528600).

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Authors and Affiliations

  1. Institute of Advanced Technology, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Umi Zulaikha Mohd Azmi, Nor Azah Yusof, Jaafar Abdullah, Shahrul Ainliah Alang Ahmad & Fatin Nabilah Mohd Faudzi

  2. Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Nor Azah Yusof, Jaafar Abdullah & Shahrul Ainliah Alang Ahmad

  3. School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor, Malaysia

    Nurul Hanun Ahmad Raston

  4. School of Medical Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia

    Siti Suraiya

  5. NanoMalaysia Berhad, a CLG under the Ministry of Energy, Science, Technology, Environment and Climate Change (MESTECC), 50450, Kuala Lumpur, Malaysia

    Poh Shing Ong & Devandran Krishnan

  6. School of Medicine, Trinity Biomedical Sciences Institute, Trinity College, Dublin 2, Ireland

    Nur Khairunnisa Sahar

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  1. Umi Zulaikha Mohd Azmi
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Correspondence to Umi Zulaikha Mohd Azmi or Nor Azah Yusof.

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Mohd Azmi, U.Z., Yusof, N.A., Abdullah, J. et al. Portable electrochemical immunosensor for detection of Mycobacterium tuberculosis secreted protein CFP10-ESAT6 in clinical sputum samples. Microchim Acta 188, 20 (2021). https://doi.org/10.1007/s00604-020-04669-x

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