Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro

Abstract

Toxoplasmosis is caused by T. gondii and can create serious health problems in humans and also worldwide economic harm. Because of the clinical and veterinary importance of toxoplasmosis, its timely and accurate diagnosis has a major impact on disease-fighting strategies. T. gondii surface antigen 1 (SAG1), an immunodominant-specific antigen, is often used as a diagnostic tool. Therefore, the aim of this study was the optimization of novel fluorescein isothiocyanate (FITC) labeling of the SAG1-specific IgY antibody to show the potential for immunofluorescence imaging of T. gondii in vitro. The specificity of IgY antibodies was controlled by an enzyme-linked immunosorbent assay (ELISA), and the concentration of the IgY antibody was detected using a spectrophotometer. The optimum incubation time and FITC concentration were determined with a fluorescence spectrometer. The obtained FITC-labeled IgY was used for marking T. gondii tachyzoites, which were cultured in vitro and viewed using light microscopy. The interaction of the fluorescence-labeled antibody and the T. gondii tachyzoites was examined with a fluorescence microscope. In this study, for the first time, a FITC-labeled anti-SAG1 IgY antibody was developed according to ELISA, fluorescence spectrometer, and fluorescence imaging of cell culture. In the future, the obtained FITC-labeled T. gondii tachyzoites’ specific IgY antibodies may be used as diagnostic tools for the detection of T. gondii infections in different samples.

Egg yolk antibodies (IgY) and their applications in human and veterinary health: A review

“…Therefore, this study revealed a potential diagnostic test for toxoplasmosis. More recently, Anti-SAG1 IgY conjugated to fluorescein isothiocyanate detected T. gondii tachyzoites in a colony, by means of immunofluorescence, and may be used to detect the protozoan in other types of samples [114].…”

Enhancing the detection of Toxoplasma gondii via an anti-SAG1 scFv-alkaline phosphatase immunoconjugate

“…Similar ascertainment was recently reported by Sert et al . in using a fluorescein isothiocyanate (FITC) labeling of the SAG1-specific chicken egg yolk antibody (IgY) to show the potential for immunofluorescence imaging of T. gondii in vitro [54]. Therefore, the recombinant conjugate scFvSG15(V L T8P)-AP presented here has the potential to be used as diagnostic tools to confirm the presence of T. gondii parasites in biological samples by means of direct immunofluorescence assay.…”

The Spectrophotometric Characteristic of Immunoglobulin Conjugates for Diagnostics of Causative Agents of Especially Dangerous Infections

“…This dye, the molecular formula of which is C 21 H 11 NO 5 S, causes a green fluorescence; it bonds covalently to biomolecules via the NCS functional group and has a high quantum yield [15,16]. The maximums of the absorption and fluorescence wavelengths are 495 and 520 nm, respectively [17]; owing to this, the background fluorescence of biological specimens is reduced to the minimum.…”