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A novel engineered label-free Zn-based MOF/CMC/AuNPs electrochemical genosensor for highly sensitive determination of Haemophilus Influenzae in human plasma samples

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Abstract

An innovative label-free DNA genosensing assay based on a direct hybridization followed by DPASV in the presence of [Fe(CN)6]4−/3− was developed for recognizing the H. influenza genome in human plasma samples. To attain this objective, Zn-based MOF was synthesized and combined with carboxymethyl cellulose (CMC), which were immobilized on the surface of Au electrode and AuNPs were immobilized on the Zn-based MOF/CMC/Au-modified electrode surface. The genosensing bio-assay provides high specificity, sensitivity, and good performance for the determination of L-fuculokinase gene from the Haemophilus influenza genome. Various characterization techniques were applied including Fe-SEM, EDS, FT-IR, and XRD for investigation of morphological features and particle size. Under optimal conditions LOD and LOQ were 1.48 fM and 3.23 fM, respectively. Moreover, a wide linear range was obtained ranging from 0.1 pM–10 nM for t-DNA. The recoveries and RSDs were 98.4–103% and 2.2–3.2, respectively. The fabricated biosensing assay presented high selective ability of one, two, and three-base mismatched sequences. In addition, negative control of the genosensing assay for investigation of the selectivity was performed by the t-DNAs of Salmonella typhimurium and Shigella flexneri bacteria. Likewise, reproducibility and repeatability of the related bio-assay were investigated. It is to be noted that the organized genosensing bio-assay can be straightforwardly reused and regenerated to assess the hybridization process.

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Funding

The authors wish to thank the financial support from the University of Tabriz, Tabriz, Iran. The authors are grateful for financial support from the Immunology Research Center, Tabriz University of Medical Sciences (Grants number: 60811).

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Authors and Affiliations

  1. Department of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, 51666 16471, Iran

    Hessamaddin Sohrabi, Mir Reza Majidi & Karim Asadpour-Zeynali

  2. Department of Biochemistry, Higher Education Institute of Rab-Rashid, Tabriz, 51664, Iran

    Fatemeh Nami

  3. Research Laboratory of Advanced Water and Wastewater Treatment Processes, Department of Applied Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, 51666-16471, Iran

    Alireza Khataee

  4. Department of Environmental Engineering, Gebze Technical University, 41400, Gebze, Turkey

    Alireza Khataee

  5. Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Ahad Mokhtarzadeh

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  1. Hessamaddin Sohrabi
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  2. Mir Reza Majidi
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Correspondence to Mir Reza Majidi or Ahad Mokhtarzadeh.

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Sohrabi, H., Majidi, M.R., Nami, F. et al. A novel engineered label-free Zn-based MOF/CMC/AuNPs electrochemical genosensor for highly sensitive determination of Haemophilus Influenzae in human plasma samples. Microchim Acta 188, 100 (2021). https://doi.org/10.1007/s00604-021-04757-6

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  • DOI: https://doi.org/10.1007/s00604-021-04757-6

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