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Efficiency of a Single well IgG, IgM and IgA Anti T. gondii Fluorimetric Assay for Pre-natal Screening for Congenital Toxoplasmosis

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Abstract

Toxoplasmosis, worldwide protozoan disease, is usually benign, except when acute disease occurs in pregnant women, resulting in fetal infection with deaths or high morbidity after birth. Treatment blocks fetal infection or damage after infection, imposing a quick and effective diagnosis. Maternal infection is mostly asymptomatic thus regular serology are the main tool for detect seroconversion and acute infection in prenatal care. Screening test for specific anti T. gondii IgG, IgM and IgA must be quick, cheaper and available for the prenatal care. Fluorescent solid phase assays appears as a good alternative as they allow one well detection of IgG and IgM aside to allow high throughput in 384 wells. Here, we standardize and analyze a single well anti-T. gondii IgG, IgM and IgA immunosorbent fluorescent assay in a large sample of a public hospital. We construct conjugates for each immunoglobulin with specific fluorophores, which allows concomitant detection in a microplate fluorimeter, with stability and reproducibility, allowing cheaper 384 wells use. Tested in our 600 mother samples from a large public hospital, they presented the same reactivity as standard routine tests, but with adequate IgM and IgA screening, as adequately standardized in house ELISA, while the design of most commercial assays give false positive results. The few TFISA positive IgG, IgM and IgA samples also had low avidity IgG, confirming recent infection. TFISA will help a screening toxoplasmosis in pregnancy program in large cities, with , allowing testing large numbers of samples at low cost and must be considered for other serological purposes.

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Data and Code Availability

Sample and their data are stored at the biorepository of the Laboratório de Protozoologia do Instituto de Medicina Tropical de São Paulo, da Faculdade de Medicina da Universidade de São Paulo. The data that support the findings of this study are not openly available due to ethical restrictions and confidentiality (Plataforma Brasil project no. 01539512.8.0000.0068). The corresponding author stored informed consent forms of each participant. Sample or data of patients with confidentiality restrictions could be available from the corresponding author upon reasonable request at hfandrad@usp.br.

Abbreviations

IFA:

Indirect immunofluorescence assay

ELISA:

Enzyme linked immunosorbent assay

TFISA:

Triple fluorescent immunosorbent assay

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Acknowledgements

We gratefully thanks of Andrea da Costa, Nahiara E.Zorgi, Camila A Carvalho, A.J. Galisteo Jr. & Luciana R. Meireles for discussions and technical advice during the project. We also thanks to Elizama C. Bezerra & Giselle P. Sartori for contacting participants for the project. JP Rodrigues was supported by fellowship from CAPES for her Ph.D. thesis in Tropical Medicine. LIMHCFMUSP(49) and CNPq indirectly supported this project.

Funding

This research was supported by Fundação de Amparo a Pesquisa no Estado de São Paulo (FAPESP), a public Brazilian funding foundation at process no. 2013/04676-9. JPR used data for her Ph.D. thesis in Tropical Medicine at Universidade de São Paulo, supported by fellowship from Coordenadoria de Apoio ao Pessoal de Ensino Superior(CAPES), also public Brazilian funding agency. LIMHCFMUSP(49) and CNPq indirectly supported this project.

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Authors and Affiliations

Authors

Contributions

J.P. Rodrigues is the main author, responsible for the design of the study, standardization an execution of assays, control data bank and stored samples. She also performed most data analysis and serological indexes determinations, and correct this manuscript. She use this data for the experimental part of her Ph.D. thesis in Tropical Medicine. HFAJr was the PhD advisor of JPR and oriented the design of the project, ethical issues, grants and funding aside to data analysis and discussing solutions for problems and writing the manuscript.

Corresponding author

Correspondence to Heitor Franco de Andrade Junior.

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Ethics Approval

Brazilian ethics committees, both at university and federal councils, approved this work. Documentation of this process were deposited in transparence site as project no. 01539512.8.0000.0068 (Plataforma Brasil site https://plataformabrasil.saude.gov.br). This ethical committee also monitored the development of the project.

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Rodrigues, J.P., Junior, H.F.d. Efficiency of a Single well IgG, IgM and IgA Anti T. gondii Fluorimetric Assay for Pre-natal Screening for Congenital Toxoplasmosis. J Fluoresc 32, 661–667 (2022). https://doi.org/10.1007/s10895-022-02892-8

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