Recombinant nucleocapsid protein is potentially an inexpensive, effective serodiagnostic reagent for infectious bronchitis virus

The nucleocapsid protein of the Gray strain of infectious bronchitis virus (IBV) is highly immunogenic and cross-reactive among various distinct serotypes. Recombinant nucleocapsid polypeptide expressed in bacteria with a histidine tag at the amino terminus has been used as antigen for developing an...

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Bibliographic Details
Published in:	Journal of virological methods Vol. 70; no. 1; pp. 37 - 44
Main Authors:	Ndifuna, Abdul, Waters, Anna K., Zhou, Minglong, Collisson, Ellen W.
Format:	Journal Article
Language:	English
Published:	London Elsevier B.V 1998 Amsterdam Elsevier New York, NY Elsevier Science B.V
Subjects:	Animals Antibodies, Viral - blood Antigens, Viral - genetics Antigens, Viral - immunology Biological and medical sciences Blotting, Western - veterinary Chickens Coronavirus Infections - diagnosis Coronavirus Infections - veterinary ELISA Enzyme-Linked Immunosorbent Assay - veterinary Escherichia coli - genetics Fundamental and applied biological sciences. Psychology IBV Immunoblotting - veterinary Infectious bronchitis virus - immunology Microbiology Nucleocapsid - genetics Nucleocapsid - immunology Poultry Diseases - diagnosis Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - immunology Recombinant nucleocapsid protein Serodiagnosis Techniques used in virology Virology Recombinant nucleocapsid protein Serodiagnosis IBV ELISA Nucleocapsid Avian infectious bronchitis virus Infection Virus Avian infectious bronchitis Coronavirus Viral disease Serological method Immunoblotting assay Diagnosis Recombinant protein Coronaviridae Veterinary Nidovirales
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Summary:	The nucleocapsid protein of the Gray strain of infectious bronchitis virus (IBV) is highly immunogenic and cross-reactive among various distinct serotypes. Recombinant nucleocapsid polypeptide expressed in bacteria with a histidine tag at the amino terminus has been used as antigen for developing an assay to detect IBV-specific antibody. This fusion protein was produced readily in bacteria and easily purified with a nickel column which bound to the histidine tag. Conditions were optimized for using these preparations for an IBV-specific ELISA. Although differences in optical densities could be detected between pre-immune and positive sera for the Ark, Mass, and Gray strains with antigen concentrations between 50 and 0.1 μg per well, the greatest differences could be detected with 3 and 1.5 μg of protein per well. Maximum differences in optical densities between pre-immune and positive sera were obtained using 2.4 μg per well of protein and sera diluted between 1:80 and 1:160. In addition, as little as 30 ng/dot of recombinant nucleocapsid consistently detected IBV-specific sera in immunoblot assays which have convenient field applications.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0166-0934 1879-0984
DOI:	10.1016/S0166-0934(97)00170-5