Detection of Infectious Bronchitis Virus and Specific Anti- Viral Antibodies Using a Concanavalin A–Sandwich–ELISA

Abstract

Concanavalin A-Sandwich ELISA (Con A-S-ELISA) was developed for the detection of infectious bronchitis virus (IBV) or chicken specific anti-viral antibodies. The antigen detection limit for the Con A-S-ELISA was 10 5,1 EID 50 /mL. Three homologous and four heterologous IBV strains were similarly detected. This assay was highly effective in detecting the virus after infected tissue homogenates were passed once in embryonated chicken eggs, showing a good agreement with virus isolation technique. The Con A-S-ELISA was also used to measure anti-IBV chicken antibodies and showed a high coefficient of correlation (r ‫؍‬ 0.85) and an agreement of k ‫؍‬ 0.80 with the commercially available Indirect-ELISA. The relative sensitivity and specificity between these two tests were, respectively, 92.86% and 95.65% with an accuracy of 93.39%. Thus, the Con A-S-ELISA proved to be able to detect alternatively homologous and heterologous IBV strains or specific chicken anti-IBV antibodies, using the Con A as capture reagent of this assay. 569

Genetic grouping of avian infectious bronchitis virus isolated in Brazil based on RT-PCR/RFLP analysis of the S1 gene

“…Seven of the poultry flocks sampled had been vaccinated with live or attenuated Mass viral strain (H120) vaccines; and three were not vaccinated; the vaccination status of the other two flocks was unknown (Table 1). The IBV isolates as well as the reference H120 IBV strain were propagated in 9-10 day old specific pathogen free (SPF) embryonated chicken eggs and identified as IBV by Con A-S-ELISA (Bronzoni et al 2005).…”

Development and application of nsp5-ELISA for the detection of antibody to infectious bronchitis virus

“…ELISA has been widely used in IBV serological profiling for its characteristics of simplicity, rapidness, sensitivity and being suitable for large-scale use. In previous established ELISA assays, the whole virion or recombinant subunits of structural proteins were used as the antigens for IBV antibody detection (Bronzoni et al, 2005;Lin et al, 2012;Gibertoni et al, 2005). In this study, we first developed a nonstructural protein nsp5-based ELISA for IBV antibody detection.…”

“…Gene sequences and phylogenetic analyses are currently applied to determine the variation of newly isolated IBV. ELISA based on whole virions as well as recombinant S1 (spike protein 1 subunit) and N proteins (Nucleoprotein) could provide a rapid and large-scale detection method for IBV infection (Bronzoni et al, 2005;Lin et al, 2012;Gibertoni et al, 2005). However, few IBV detection methods are developed based on the nonstructural proteins (nsps).…”

Exploration of the metal coordination region of concanavalin A for its interaction with human norovirus

“…Concanavalin A (Con A), a lectin from jack bean, binds specifically to the non-reducing mannosyl and glycosyl residues of polysaccharides, glycoproteins, and glycolipids. Preliminary reports have demonstrated that Con A can bind to a broad range of enveloped viruses, including varicella-zoster, infectious bronchitis, and dengue viruses by interacting with the glycoproteins present on the viral surface [21,22].…”