Amperometric enzyme immunoassay sensors in determination of Fusarium oxysporum antigens
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Amperometric enzyme immunoassay sensors in determination of Fusarium oxysporum antigens

R.M. Beilinson a*, E.P. Medyantseva a**, E.A. Kirillova a***, E.V. Khaldeeva b****, H.C. Budnikov a*****

aKazan Federal University, Kazan, 420008 Russia

bKazan Research Institute of Epidemiology and Microbiology, Kazan, 420015 Russia

E-mail: *rvarlamo@mail.ru, **emedyant@gmail.com, ***elena.al.kirillova@gmail.com,

****mycology-kazan@yandex.ru, *****Herman.Budnikov@kpfu.ru

Received September 24, 2021


ORIGINAL ARTICLE

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DOI: 10.26907/2542-064X.2022.1.22-35

For citation: Beilinson R.M., Medyantseva E.P., Kirillova E.A., Khaldeeva E.V., Budnikov H.C. Amperometric enzyme immunoassay sensors in determination of Fusarium oxysporum antigens. Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki, 2022, vol. 164, no. 1, pp. 22–35. doi: 10.26907/2542-064X.2022.1.22-35. (In Russian)

Abstract

Enzyme immunoassay sensors (EIS) with L-cysteine desulfhydrase (CDH) as a label based on planar graphite electrodes were created to determine Fusarium oxysporum antigen (Ag). The EIS action relies on the combined effect of immunological, enzymatic, and electrochemical reactions. The immunosensors developed allow the determination of Fusarium oxysporum Ag in the concentration range of 5?10–10 – 5?10–6 mg/mL with the limit of quantification of 1?10–10 mg/mL. The binding constant of the Ab-Ag immune complexes was estimated as Ка1 = (6.2 ? 0.2)?109 (mg/mL)–1. No cross-reaction with Ag of such pathogenic fungi as Alternaria alternata, Cladosporium herbarum, Aspergillus fumigatus, and Aspergillus niger was confirmed. Here the quality of wheat groats was assessed to test the methods for the determination of Fusarium oxysporum Ag using the developed CDH immunosensor.

Keywords: Fusarium oxysporum, enzyme immunoassay sensor, L-cysteine desulfhydrase, fungal pathogens

Figure Captions

Fig. 1. Schematic representation of the action of enzyme immunoassay sensors based on immobilized Ab against Fusarium oxysporum Ag and the enzyme (E) includes: co-immobilized enzyme and Ab (a), Ag in solution (b), immune complex that is formed and variants of the substrate access to the active enzyme surface (c).

Fig. 2. Voltammograms of the electrooxidation of a 1?10–3 M cysteine solution in phosphate buffer (pH 7.6) (1), in the presence of L-cysteine desulfhydrase (2), in the absence of L-cysteine desulfhydrase (4), in the presence of Fusarium oxysporum Ag and L-cysteine desulfhydrase (3) at the planar graphite electrode.

Fig. 3. Scatchard plot for determining the binding constant of the Ab-Ag immune complex of Fusarium oxysporum.

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