Development of a novel immunoFET technology-based POC assay for detection of Leishmania donovani and Leishmania major.

Publisher: Wiley Country of Publication: England NLM ID: 7910948 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1365-3024 (Electronic) Linking ISSN: 01419838 NLM ISO Abbreviation: Parasite Immunol Subsets: MEDLINE

Original Publication: Oxford, Wiley

Leishmania donovani* ; Leishmania major* ; Leishmaniasis*/parasitology; Humans ; Point-of-Care Systems ; Ribosomal Proteins ; Antibodies, Protozoan ; Neglected Diseases

Leishmaniasis is considered as one of the 20 neglected tropical diseases. Current methods of leishmanial diagnosis depend on conventional laboratory-based techniques, which are time-consuming, costly and require special equipment and trained personnel. In this context, we aimed to provide an immuno field effect transistors (ImmunoFET) biosensor that matches the conventional standards for point-of-care (POC) monitoring and detection of Leishmania (L.) donovani/Leishmania major. Crude antigens prepared by repeated freeze thawing of L. donovani/L. major stationary phase promastigotes were used for ELISA and ImmunoFETs. Lesishmania-specific antigens were serially diluted in 1× PBS from a concentration of 10 ⁶ -10 ² parasites/mL. A specific polyclonal antibody-based sandwich ELISA was established for the detection of Leishmania antigens. An immunoFET technology-based POC novel assay was constructed for the detection of Leishmania antigens. Interactions between antigen-antibody at the gate surface generate an electrical signal that can be measured by semiconductor field-effect principles. Sensitivity was considered and measured as the change in current divided by the initial current. The final L. donovani/L. major crude antigen protein concentrations were measured as 1.50 mg/mL. Sandwich ELISA against the Leishmania 40S ribosomal protein detected Leishmania antigens could detect as few as 100 L. donovani/L. major parasites. An immunoFET biosensor was constructed based on the optimization of aluminium gallium nitride/gallium nitride (AlGaN/GaN) surface oxidation methods. The device surface was composed by an AlGaN/GaN wafer with a 23 nm AlGaN barrier layer, a 2 μm GaN layer on the silicon carbide (SiC) substrate for Leishmania binding, and coated with a specific antibody against the Leishmania 40S ribosomal protein, which was successfully detected at concentrations from 10 ⁶ to 10 ² parasites/mL in 1× PBS. At the concentration of 10 ⁴ parasites, the immunoFETs device sensitivities were 13% and 0.052% in the sub-threshold regime and the saturation regime, respectively. Leishmania parasites were successfully detected by the ImmunoFET biosensor at a diluted concentration as low as 150 ng/mL. In this study, the developed ImmunoFET biosensor performed well. ImmunoFET biosensors can be used as an alternative diagnostic method to ELISA. Increasing the sensitivity and optimization of immuno-FET biosensors might allow earlier and faster detection of leishmaniasis.
(© 2023 John Wiley & Sons Ltd.)

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The Scientific and Technological Research Council of Türkiye

Keywords: Leishmania species; biosensor; immuno field effect transistors; leishmaniasis; point-of-care

0 (aluminum gallium nitride)
0 (Ribosomal Proteins)
0 (Antibodies, Protozoan)

Date Created: 20230515 Date Completed: 20230626 Latest Revision: 20231116

20240829

10.1111/pim.12984

37183939