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Use of polyclonal IgY antibodies to detect serum immune complexes in patients with active hookworm infection

Published online by Cambridge University Press:  13 February 2020

Dayane Costa de Souza ,
Lucas Silva de Faria ,
José Eduardo Neto de Sousa ,
Camila de Almeida Lopes ,
Vanessa da Silva Ribeiro ,
Vivian Jordania da Silva ,
Raphaella Paula Ribeiro ,
Élida Mara Leite Rabelo ,
Veridiana Melo Rodrigues Ávila  and
Álvaro Ferreira Júnior
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Dayane Costa de Souza
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Av. Amazonas s/n, bloco 4C, Uberlândia, Minas Gerais, 38400-902, Brasil
Lucas Silva de Faria
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Av. Amazonas s/n, bloco 4C, Uberlândia, Minas Gerais, 38400-902, Brasil
José Eduardo Neto de Sousa
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Av. Amazonas s/n, bloco 4C, Uberlândia, Minas Gerais, 38400-902, Brasil
Camila de Almeida Lopes
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Av. Amazonas s/n, bloco 4C, Uberlândia, Minas Gerais, 38400-902, Brasil
Vanessa da Silva Ribeiro
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Av. Amazonas s/n, bloco 4C, Uberlândia, Minas Gerais, 38400-902, Brasil
Vivian Jordania da Silva
Affiliation:
Laboratório de Parasitologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, Minas Gerais, 31270-901, Brasil
Raphaella Paula Ribeiro
Affiliation:
Programa de Pós-Graduação em Sanidade e Produção Animal nos Trópicos, Universidade de Uberaba – Uniube, Campus Aeroporto. Av. Nenê Sabino, sala 2D05, Uberaba, Minas Gerais, 38055-500, Brasil
Élida Mara Leite Rabelo
Affiliation:
Laboratório de Parasitologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, Minas Gerais, 31270-901, Brasil
Veridiana Melo Rodrigues Ávila
Affiliation:
Laboratório de Bioquímica e Toxinas Animais, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia – UFU, Av. Pará 1720, Uberlândia, Minas Gerais, 38400-902, Brasil
Álvaro Ferreira Júnior
Affiliation:
Programa de Pós-Graduação em Sanidade e Produção Animal nos Trópicos, Universidade de Uberaba – Uniube, Campus Aeroporto. Av. Nenê Sabino, sala 2D05, Uberaba, Minas Gerais, 38055-500, Brasil
Julia Maria Costa-Cruz*
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Av. Amazonas s/n, bloco 4C, Uberlândia, Minas Gerais, 38400-902, Brasil
*
Author for correspondence: Julia Maria Costa-Cruz, E-mail: costacruz@ufu.br
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Abstract

Definitive diagnosis of hookworm infection is usually based on the microscopic detection of eggs in a stool sample; however, several cases display a low or irregular egg output. Serodiagnosis can be a useful tool to identify these cases, but conventional tests do not differentiate past from active infections. The aim of this study was to obtain and apply egg yolk polyclonal immunoglobulin (IgY) antibodies to detect immune complexes (ICs) in serum samples from patients infected with hookworm. Hens were immunized with Ancylostoma ceylanicum saline extract, their eggs were collected and then IgY antibodies were extracted and purified. Antibody purity was tested by 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis and specificity was assessed by immunoblotting and immunofluorescence. IgY production was evaluated by kinetics enzyme-linked immunosorbent assay (ELISA). Sandwich ELISA tested the ability of IgY to detect ICs in serum samples, from which diagnostic parameters were calculated. Antibody responses increased steadily from day 7 to 42. In the immunoblotting assay, IgY recognized two protein complexes. The immunofluorescence assay showed no staining in control samples. The sandwich ELISA presented a very high diagnostic value, with a sensitivity of 90% and a specificity of 86.7%. Our pioneer strategy highlights the potential use of egg yolk IgY as a diagnostic test to detect active hookworm infection.

Keywords

Active infectionhookwormimmune complexesIgY
Type
Research Article
Information
Parasitology , Volume 147 , Issue 6 , May 2020 , pp. 715 - 720
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Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

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