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Anti-Ascaris suum immunoglobulin Y as a novel biotechnological tool for the diagnosis of human ascariasis

Published online by Cambridge University Press:  14 August 2019

C.A. Lopes
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
L.S. de Faria
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
J.E.N. de Sousa
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
I.P. Borges
Affiliation:
Laboratório de Bioquímica e Toxinas Animais, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
R.P. Ribeiro
Affiliation:
Programa de Pós-Graduação em Sanidade e Produção Animal nos Trópicos, Universidade de Uberaba, Campus Aeroporto. Av. Nenê Sabino, 1802, sala 2D05, Uberaba, Minas Gerais 38055-500, Brazil
L.L. Bueno
Affiliation:
Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Belo Horizonte, Minas Gerais 31270-901, Brazil
V.M. Rodrigues Ávila
Affiliation:
Laboratório de Bioquímica e Toxinas Animais, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
Á. Ferreira Júnior
Affiliation:
Programa de Pós-Graduação em Sanidade e Produção Animal nos Trópicos, Universidade de Uberaba, Campus Aeroporto. Av. Nenê Sabino, 1802, sala 2D05, Uberaba, Minas Gerais 38055-500, Brazil
J.M. Costa-Cruz*
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
*
Author for correspondence: J.M. Costa-Cruz, E-mail: [email protected]

Abstract

Human ascariasis is a neglected tropical disease of great relevance to public health and is considered the most frequent helminthiasis in poor regions. Accurately diagnosing this parasite has been challenging due to limitations of current diagnostic methods. Immunoglobulin Y (IgY) technology is a very effective alternative for the production of highly specific and profitable antibodies. This study aimed to produce and apply anti-Ascaris suum IgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract from A. suum adult life forms were given at 14-day intervals to Gallus gallus domesticus hens of the Isa Brown line. Eggs and blood samples were collected weekly and fortnightly, respectively, to monitor the production of antibodies. The specificity of antibodies was confirmed by dot-blot, kinetic enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence antibody tests. The application for disease diagnosis was performed through the detection of immune complexes in human serum samples by sandwich ELISA. Peaks of IgY anti-A. suum production occurred at weeks 6 and 8. IgY showed high avidity levels after the second dose of immunization, ranging from 64% to 93%, with a mean avidity index of 78.30%. Purified IgY recognized 12 bands of proteins from A. suum saline extract. Eggs, the uterine portion and cuticles of A. suum female adult are reactive in immunofluorescence. The detection of immune complexes showed diagnostic values of 80% sensitivity and 90% specificity. In conclusion, specific IgY have been shown to be a potential immunodiagnostic tool with promising future applications in human ascariasis.

Type
Research Paper
Copyright
Copyright © Cambridge University Press 2019 

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