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Anal Biochem. 2019 Jan 1;564-565:116-122. doi: 10.1016/j.ab.2018.10.025. Epub 2018 Oct 28.

Serological diagnosis of Toxoplasmosis disease using a fluorescent immunosensor with chitosan-ZnO-nanoparticles.

Author information

1
INQUISAL, Departamento de Química, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis, Chacabuco 917, San Luis, Argentina.
2
Instituto Tecnológico de Chascomús (IIB-INTECH) CONICET-Universidad Nacional de General San Martín, Camino de Circunvalación Laguna Km. 6 CC 164, B7130IWA, Chascomús, Provincia de Buenos Aires, Argentina.
3
INQUISAL, Departamento de Química, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis, Chacabuco 917, San Luis, Argentina. Electronic address: messina@unsl.edu.ar.

Abstract

This article describes a microfluidic LIF immunosensor for the quantitative determination of anti-Toxoplasma gondii IgG (anti-T. gondii) specific antibodies. The serological detection of these antibodies plays a crucial role in the clinical diagnosis of toxoplasmosis. Zinc oxide nanoparticles (ZnO-NPs) obtained by wet chemical procedure were covered with chitosan and then used to conjugate T-gondii antigens into the central microfluidic channel. Serum samples containing anti-T-gondii IgG antibodies were injected into the immunosensor where they interact immunologically with T. gondii antigens. Bound antibodies were quantified by the addition of anti-IgG antibodies labeled whit alkaline phosphatase (ALP). ALP enzymatically converts the non-fluorescent 4-methylumbelliferyl phosphate (4-MUP) to soluble fluorescent methylumbelliferone that was measured using excitation at 355 nm and emission at 440 nm. The relative fluorescent response of methylumbelliferone is proportional to the concentration of anti-T. gondii IgG antibodies. The coefficients of variation are less than 4.73% for within-day assays and less than 6.34% for between-day assays. Results acquired by LIF immunosensor agree with those obtained by enzyme-linked immunosorbent assay method, suggesting that the designed sensor represents a promising tool for the quantitative determination of anti-T. gondii IgG antibodies of clinical samples.

KEYWORDS:

ALP; CH-ZnO-NPs; Immunosensor; Laser-induced fluorescence Toxoplasma gondii

PMID:
30393087
DOI:
10.1016/j.ab.2018.10.025

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