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A Colloidal Gold Immunochromatographic Strip Based on a Conserved Epitope Peptide for Rapid Detection of Antibodies
Ling Liu1, Kang Zhao1, Chang-Run Zhao1
1College of Animal Science and Technology, Guangxi University, Nanning 530004, China.
Abstract:
Avian infectious bronchitis virus (IBV) is widely distributed worldwide and causes substantial economic losses to the poultry industry. Because IBV undergoes frequent mutation, prevention and control of infection remain challenging. Immunization is an important measure for the prevention and control of IB. Therefore, there is an urgent need for a rapid, sensitive, specific, and convenient method for the detection of antibodies against IBV. In this study, we firstly developed an indirect colloidal gold immunochromatographic strip for the rapid detection of antibodies against IBV based on a conserved epitope peptide. The recombinant epitope peptide recognized by N2D5 monoclonal antibody (mAb) against the N protein of IBV was expressed as a GST fusion protein (GST-N2D5) based on the conserved antigenic epitope previously identified in our laboratory. Colloidal gold-labeled GST-N2D5 was used as the detection reagent to generate visual signals. Rabbit anti-chicken IgY and mouse anti-GST mAb were immobilized on the nitrocellulose membrane as the test line (T line) and control line (C line), respectively. The optimal pH and optimal protein concentration for conjugation of gold nanoparticles (AuNPs) with GST-N2D5 were pH 8.5 and 72 µg/mL, respectively. Specificity was evaluated using common avian pathogens, and no cross-reactivity was observed. The detection limit of the strip for IBV-positive serum was 1:180. In addition, the assay showed good reproducibility and stability, and results could be observed within 5 min without any specialized equipment. Clinical chicken serum samples were tested using both the developed strip and an enzyme-linked immunosorbent assay (ELISA), and the strip showed high agreement with the ELISA. In conclusion, the established immunochromatographic strip is rapid, sensitive, specific, and easy to operate, and therefore has potential as an on-site tool for the rapid detection of antibodies against IBV, particularly in resource-limited settings.
