Influenza A viral nucleoprotein detection in isolates from human and various animal species

• Published in: Archives of virology Vol. 100; no. 1/2; pp. 75 - 87
• Main Authors: Siebinga, J.T, De Boer, G.F
• Format: Journal Article
• Language: English
• Published: Wien Springer 01-01-1988; New York, NY
• Subjects: Allantois - microbiology; Animals; Antibodies, Monoclonal; antigens; Biological and medical sciences; Chickens; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Fundamental and applied biological sciences. Psychology; genome; hemagglutination; Hemagglutination Tests; horses; Humans; influenza A virus; Influenza A virus - analysis; Influenza A virus - isolation & purification; Influenza in Birds - diagnosis; influenzavirus; Lung - microbiology; Lung - pathology; Microbiology; nucleoproteins; Nucleoproteins - analysis; Orthomyxoviridae Infections - diagnosis; Orthomyxoviridae Infections - pathology; RNA; Swine; Techniques used in virology; Viral Proteins - analysis; Virology; Virus; Human origin; Influenzavirus A; Hemagglutination test; Animal origin; Orthomyxoviridae; Identification; Monoclonal antibody; Influenzavirus; ELISA assay; Nuclear protein; Immunological method
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/BF01310909

A double antibody sandwich, enzyme-linked immunosorbent assay (DAS-ELISA) was developed to detect influenza A viral antigen, employing a monoclonal antibody directed against type-specific influenza A nucleoprotein (McAb anti-NP). McAb anti-NP was used to coat ELISA plates as well as to prepare the peroxidase conjugate. Influenza A viruses of avian, equine, swine, and human origin were detected in allantoic fluids of inoculated eggs with higher sensitivity by the DAS-ELISA than by hemagglutination (HA) assays. Minimal concentrations of 8 ng/ml influenza virus protein were detected in Nonidet P40-treated virus preparations. Viral antigen detection in tissues of experimentally infected chickens and pigs was successful, but in pigs yielded a lower positive score than the conventional method of virus isolation in eggs. The test is sensitive, rapid, and easy to perform, but does not permit influenza A subtyping. In avian species, the McAb anti-NP DAS-ELISA differentiates between influenza and Newcastle disease viruses. In pigs, the test distinguishes between influenza and Aujeszky's disease.