Background Infectious 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. Results We 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. Conclusions Two 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.
【저자키워드】 Antibody detection, Infectious bronchitis, Protein chip, 【초록키워드】 Anemia, vaccination, antibody, Variation, Infection, Test, Diagnosis, Influenza virus, virus, ELISA, Protein, enzyme-linked immunosorbent assay, Outbreaks, pathogen, Surveillance, Sensitivity and specificity, RDT, Rapid diagnostic test, serological assays, Microarray, chemiluminescent immunoassay, diagnostic methods, disease, Critical, positive rate, ELISAs, newcastle disease virus, Immune status, early infection, Efficiency, Escherichia coli, bronchitis, coefficient, reproducibility, subtype, IBV, Result, detect, caused, indicated, assays, faster, expressed, purified, ranged, CIT, 【제목키워드】 antibody, Protein, novel, Infectious Bronchitis Virus,