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

Parasitology 2020 May

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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.

Citation

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, Álvaro Ferreira Júnior, Julia Maria Costa-Cruz. Use of polyclonal IgY antibodies to detect serum immune complexes in patients with active hookworm infection. Parasitology. 2020 May;147(6):715-720