Abstract
Infectious bronchitis virus (IBV) causes substantial loss to the poultry industry despite extensive vaccination. Assessing the antibody response is important for the development and evaluation of effective vaccines. We have developed an enzyme-linked immunosorbent assay (ELISA) for the detection of IBV-specific antibodies, using a synthetic peptide based on a conserved sequence in the IBV spike protein. This peptide-based ELISA (pELISA) specifically detects antibodies to different genotypes of IBV but not antibodies against other common chicken viruses. This assay could detect IBV-specific antibody response on as early as day 7 postinfection. In the testing with field serum samples collected from chickens administered with IBV vaccines, the sensitivity, specificity, and accuracy of pELISA were 98.30, 94.12, and 98.8%, respectively, relative to indirect immunofluorescence assay. Our data demonstrate that the pELISA is of value for the detection of IBV antibody and the evaluation of IBV vaccines.
Highlights
Avian infectious bronchitis, a highly contagious disease, is caused by a coronavirus, that is, infectious bronchitis virus (IBV)
Serum samples that were used in our study included 100 serum samples collected from specific-pathogen-free (SPF) chickens (Spirax Ferrer Poultry Science and Technology Co., Ltd., Jinan, China), 250 serum samples collected from chickens that were vaccinated with IBV vaccines H120 and H52 (Lihua Animal Husbandry Co., LTD, Jiangsu, China), and sera against IBV strains Massachusetts 41 (M41), 4/91, H52, H120, and CK/CH/2010/JT1, which were prepared in our laboratory by infecting SPF chickens with 1,000 median egg infectious dose (EID50) of each strain
To optimize the peptide-based Enzyme-linked immunosorbent assays (ELISAs) (pELISA), two-fold serial dilutions of the synthetic peptide and different dilutions of the positive and negative chicken serum samples, which had been confirmed by immunofluorescence assay (IFA), were tested
Summary
A highly contagious disease, is caused by a coronavirus, that is, infectious bronchitis virus (IBV). The IBV genome encodes four structural proteins as well as at least 15 non-structural and accessory proteins [1]. Among these proteins, the surface spike (S) glycoprotein is the major antigen that induces protective immune response against IBV [3]. The S2 subunit is more conserved than S1 and plays a role in inducing protective immune response [5,6,7], as well as facilitating membrane fusion and viral entry [5, 8, 9]. It has been reported that S2 could produce cross-protection against strains that differ in their S1 subunits [7]
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