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Detection of IgG Anti-Giardia duodenalis Antibodies in Sera by Indirect Immunofluorescence and Western Blotting Assays

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Abstract

Introduction

Serological assays are alternative laboratory tools for the diagnosis of parasitic infections. The aim of this work was to evaluate the performance of the indirect fluorescent antibody test (IFAT) and Western blotting (WB) for the detection of IgG anti-Giardia antibodies in human sera.

Methodology

Sera from individuals infected with Giardia duodenalis, other parasites or non-parasitized were selected for serological assays. Ninety-seven sera were tested by IFAT at 1:20 and 1:40 dilutions and of these, 40 samples were also analyzed by WB.

Results

The sensitivity and specificity of the IFAT was 97% and 46.9% at 1:20 sera dilution, and 39.4% and 59.4% at 1:40 sera dilution. The low molecular weight polypeptides fractions of 25 kDa, 27–31 kDa and 45–55 kDa were the most frequently identified by the sera of individuals infected with G. duodenalis, along with low cross-reactivity, presenting an individual sensitivity of 42.8%, 50.0% and 57.1%, and specificity of 83.3%, 83.3% and 91.7%, respectively. The highest overall sensitivity of WB (85.7%) was based on the immunoreactivity of sera with at least one of those proteins. The concordance between the detection of G. duodenalis in feces by microscopy and the WB results was considered substantial (Kappa = 0.61).

Conclusion

Constant exposure to Giardia infection throughout a lifetime can maintain high levels of specific antibodies in serum, even without active infection. Moreover, proteins found in intestinal amoebas may hinder the serological diagnosis of giardiasis in endemic areas due to cross-reactivity, which can be partially solved using Giardia low molecular weight proteins.

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Funding

This work was supported by the Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB—SUS0024/2018) and with graduation and post-graduation scholarships by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/MCT).

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Authors and Affiliations

  1. Programa de Pós-Graduação em Farmácia, Faculdade de Farmácia, Universidade Federal da Bahia, Salvador, Bahia, Brazil

    Samara Alves Santos, Neci Matos Soares & Márcia Cristina Aquino Teixeira

  2. Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Universidade Federal da Bahia, Salvador, Bahia, 40170-115, Brazil

    Joelma Nascimento de Souza, Mariana Conceição Santos, Darleide dos Santos Novais, Victoria Nascimento Suzart, Isabela dos Santos Guedes, Maena Honda Neves, Neci Matos Soares & Márcia Cristina Aquino Teixeira

  3. Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia, Salvador, Bahia, Brazil

    Flávia Thamiris Figueiredo Pacheco

  4. Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brasil

    Maria Aparecida Gomes

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  1. Samara Alves Santos
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Contributions

SAS: made substantial contributions to the acquisition and interpretation of data; wrote the manuscript with support of MCAT. FTFP, MCS, DdSN, VNS, IdSG, MHN, and MAG contributed to the acquisition and analysis of data for the work. JNdS and NMS: aided in interpreting the results; contributed to manuscript draft and revised it critically for scientific content. MCAT: designed the study and supervised the work; made substantial contributions to the interpretation of data for preparation of the manuscript and revised it critically for scientific content.

Corresponding author

Correspondence to Márcia Cristina Aquino Teixeira.

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The Committee of Ethics in Research of the Pharmacy College, Federal University of Bahia, Brazil, approved this study, under the registry number 1.904.340. The Ethics Committee of Nursing School, Federal University of Bahia, Brazil, approved the study (project approval No. 907.867).

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Santos, S.A., de Souza, J.N., Pacheco, F.T.F. et al. Detection of IgG Anti-Giardia duodenalis Antibodies in Sera by Indirect Immunofluorescence and Western Blotting Assays. Acta Parasit. 69, 351–357 (2024). https://doi.org/10.1007/s11686-023-00753-3

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