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Impedimetric Biosensor for Bovine Herpesvirus Type 1‐Antigen Detection
Author(s) -
Garcia Luane Ferreira,
Silvio Batista Rodrigues Edson,
Rocha Lino de Souza Guilherme,
Jubé Wastowski Isabela,
Mota de Oliveira Fernando,
Torres Pio dos Santos Wallans,
Souza Gil Eric
Publication year - 2020
Publication title -
electroanalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 128
eISSN - 1521-4109
pISSN - 1040-0397
DOI - 10.1002/elan.201900606
Subject(s) - bovine herpesvirus 1 , antigen , biosensor , detection limit , antibody , dielectric spectroscopy , nasal administration , immune system , pathogen , chromatography , chemistry , virology , biology , electrode , microbiology and biotechnology , immunology , electrochemistry , virus , herpesviridae , biochemistry , viral disease
The bovine herpesvirus type 1 (BHV‐1) is a pathogen of great economic impact for livestock, which is related multi‐systemic infections that leads to mortality or morbidity of cattles. Thus, the search for cheap and practical methodologies that allow the selective detection of BHV‐1 antigen (BHV‐1 AG) is of utmost relevance. Therefore, an impedimetric label‐free immunosensor was herein, developed and its performance evaluated in biological samples enriched with BHV‐1 AG. Briefly, the biosensor construction was based on the immobilization of BHV‐1 antibody (BHV‐1 AB) and casein on the activated glassy carbon electrode surface. The BHV‐1 AB was isolated from egg yolk of immunized chickens, which is a less stressful protocol. The bio sensing principle was based on Electrochemical Impedance Spectroscopy by using Fe(CN) 6 4−/3− probe, which were also used to check variation of charge transfer resistance (▵ R ct ), when the electrode surface was increasingly blocked by immune complex. A linear relationship between ▵ R ct and BHV‐1 AG concentration was verified in the range from 10 to 50 TCID 50 mL −1 , with LOQ of 2.00 TCID 50 mL −1 and LOD of 0.66 TCID 50 mL −1 . Besides the suitable sensitivity, the immunosensor displayed accuracy, stability, and specificity to detect BHV‐1 AG in biological samples of serum, nasal secretions, semen and urine. Moreover, to the best of our knowledge this is the first immunosensor applied to BHV‐1 diagnostic.