A diagnostic immunoassay for Newcastle disease virus based on the nucleocapsid protein expressed by a recombinant baculovirus.
A recombinant baculovirus containing a cDNA clone encoding the nucleocapsid (NP) protein of Newcastle disease virus (strain Ulster 2C) has been used to infect insect cells (Spodoptera frugiperda). High levels of overexpressed NP protein were observed, comprising up to 40% of total cellular protein, which were subsequently shown to be antigenic. Nucleoprotein derived from the crude soluble lysate of infected insect cells has been used in an indirect ELISA to detect the presence of anti-NDV antibodies in a cohort of chicken sera. Data produced from these tests indicated a good correlation between ELISA titre and haemagglutination inhibition test data. The test was not affected by interference from background cellular proteins nor by cross-reactivity with non-NDV poultry pathogens. Additionally, the test did not generate false-positive readings.
Errington
W
W
Department of Biochemistry and Genetics, Medical School, University of Newcastle upon Tyne, UK.
Steward
M
M
Emmerson
P T
PT
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005069:Evaluation Studies as Topic; D005822:Genetic Vectors / Q000235:genetics; D006385:Hemagglutination Inhibition Tests; D007074:Immunoglobulin G / Q000276:immunology; D009521:Newcastle Disease / Q000175:diagnosis; D009522:Newcastle disease virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011201:Poultry Diseases / Q000175:diagnosis / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D018411:Spodoptera / Q000166:cytology; D014758:Viral Core Proteins / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins