Novel protein chip for the detection of antibodies against infectious bronchitis virus

Abstract

BackgroundInfectious bronchitis (IB) caused by the IB virus (IBV) can cause acute damage to chickens around the world. Therefore, rapid diagnosis and immune status determination are critical for controlling IBV outbreaks. Enzyme-linked immunosorbent assays (ELISAs) have been widely used in the detection of IBV antibodies in the early infection and continuous infection of IB because they are more sensitive and quicker than other diagnostic methods.ResultsWe have developed two indirect microarray methods to detect antibodies against IBV: a chemiluminescent immunoassay test (CIT) and a rapid diagnostic test (RDT). IBV nonstructural protein 5 (nsp5) was expressed, purified from Escherichia coli, and used to spot the initiator integrated poly(dimethylsiloxane), which can provide a near “zero” background for serological assays. Compared with the IDEXX IBV Ab Test kit, CIT and RDT have a sensitivity and specificity of at least 98.88% and 91.67%, respectively. No cross-reaction was detected with antibodies against avian influenza virus subtypes (H5, H7, and H9), Newcastle disease virus, Marek’s disease virus, infectious bursal disease virus, and chicken anemia virus. The coefficients of variation of the reproducibility of the intra- and inter-assays for CIT ranged from 0.8 to 18.63%. The reproducibility of RDT was consistent with the original results. The application of the IBV nsp5 protein microarray showed that the positive rate of the CIT was 96.77%, that of the nsp5 ELISA was 91.40%, and that of the RDT was 90.32%. Furthermore, the RDT, which was visible to the naked eye, could be completed within 15 min. Our results indicated that compared with nsp5 ELISA, the CIT was more sensitive, and the RDT had similar positive rates but was faster. Furthermore, the two proposed methods were specific and stable.ConclusionsTwo microarray assays, which were rapid, specific, sensitive, and relatively simple, were developed for the detection of an antibody against IBV. These methods can be of great value for the surveillance of pathogens and monitoring the efficiency of vaccination.

Infectious Bronchitis Virus Evolution, Diagnosis and Control

“…Aside from the described methods, other serological assays have been recently developed, including methods based on Unknown multi-analyte profiling (xMap) [ 61 ], microarray [ 62 ] and strip-based [ 63 ] technologies, but they are not commercially available yet.…”

Multiplex gradient immunochip for detection of post-vaccinal antibodies in poultry

“…Some new methods based on microarrays for anti-pathogen chicken antibodies detection are proposed, however, the number of papers regarding this topic is minimal. To detect antibodies to the chicken infectious bursal disease virus, a membrane microchip with chemiluminescent detection was developed (Yan et al 2018).…”

Efficacy of Recombinant S1 Protein Expressed in Pichia pastoris in Chicken & Using for Detection Titer Antibodies against Avian Infectious Bronchitis by ELISA

“…It encodes four major structural proteins, named, glycosylated spike protein (S), membrane protein (M), phosphorylated nucleoprotein (N), and envelope protein (E), and 15 nonstructural proteins (nsp2-nsp16). Generally, nonstructural proteins are present in infected cells but not in the virus, and they only play a role in the process of virus infection and replication [6].…”