An ELISA for antibodies to infectious bronchitis virus based on nucleocapsid protein produced in Escherichia coli
نویسنده
چکیده
The nucleocapsid (N) gene of infectious bronchitis virus (IBV) strain X isolated in China was expressed in E. coli and was purified as a recombinant protein. An indirect ELISA assay (N-ELISA) for antibody detection was established using the purified recombinant nucleocapsid protein. Antigen coating conditions and serum dilution for the N-ELISA were optimized. The S/P ratio of the absorbency value was calculated in the N-ELISA to evaluate the antibody level of chicken serum. In an experiment to test field samples for antibody detection, the N-ELISA assay shared 95.7% identity of total positive ratio with the commercial ELISA kit. It indicated that the N-ELISA assay, which was safer and easier to prepare than traditional methods, was a good candidate for evaluation of IB vaccine efficiency and virus exposure.
منابع مشابه
Development of a recombinant protein-based dot-blot hybridization assay for the detection of antibody to chicken infectious bronchitis virus
Nucleocapsid (N) protein of infectious bronchitis virus (IBV), one of the viral structural proteins, inducesstrong antibody response in natural infection. In this study, a simple, recombinant N protein-based dot-blottest was developed to serologically examine chicken serum samples for the presence of IBV antibody.Initially, 72 serum samples were tested for the presence of IBV antibody using a c...
متن کاملMolecular analysis of the nucleocapsid gene and 3' untranslated region of two infectious Bronchitis Virus field isolates from Iranian poultry farms
Infectious bronchitis (IB) is an economically important disease of chickens. Due to the emergence of new variants of infectious bronchitis virus (IBV), the control of IB has become a serious problem for the poultry industry worldwide. In the present study, the nucleocapsid gene (N) and 3' untranslated region (UTR) of two IBVs isolated from Iranian poultry farms were sequenced and compared with ...
متن کاملMolecular Cloning and Expression of Nucleocapsid Gene of Chicken Infectious Bronchitis Virus Strain Massachusetts H120
Chicken infectious bronchitis virus (IBV) causes severe respiratory and renal dysfunction in chickens. In the present study, we used reverse transcription-polymerase chain reaction (RT-PCR) to amplify the nucleocapsid (N) protein gene of strain Massachusetts (Mass) H120 of IBV that is commonly used in the vaccine production in Iran. The PCR product with the expected size of 1.2 kb was cloned in...
متن کاملCloning and expression of fragment of the rabies virus nucleoprotein gene in Escherichia coli and evaluation of antigenicity of the expression product
Rabies virus nucleoprotein (N protein) encapsidates genomic RNA of the virus and forms the viral ribonucleoprotein complex. These N proteins represent highly organized structures which activate proliferation of B cells and production antibodies against the N protein. In addition to the B cell, the rabies virus N protein has been shown to induce potent T helper cell responses resulting in a long...
متن کاملDevelopment and application of a Saccharomyces cerevisiae-expressed nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibodies against infectious bronchitis virus.
A Saccharomyces cerevisiae-expressed nucleocapsid (N) polypeptide of the M41 strain of infectious bronchitis virus (IBV) was used as antigen in a recombinant yeast-expressed N protein-based enzyme-linked immunosorbent assay (Y-N-ELISA). The Y-N-ELISA was rapid, sensitive, and specific for detecting chicken serum antibodies to IBV, and it compared favorably with a commercial ELISA.
متن کامل