ABSTRACT
Avian Influenza A virus (AIV) subtype H5 is divided into American and Eurasian lineages, according to hemagglutinin gene sequences. Although methods for detecting H5 AIVs have been described, no H5 strain-specific detection method has been reported. The purpose of the present study was to develop an antigen-capture enzyme-linked immunosorbent assay (ACE) to detect and differentiate between the American and the Eurasian H5 AIVs. Monoclonal antibodies (mAbs) against the HA fragment of a Eurasian H5N2 AIV were used as the capture antibodies as well as the detector antibodies after labeling with horseradish peroxidase to develop an ACE. One mAb was selected for detecting the American as well as the Eurasian H5 AIVs. The other mAb was used for detecting only the Eurasian H5N2 but not the American H5N2 AIVs, H6N1 AIVs, or Newcastle disease virus. The ACEs developed would be useful for detection and differentiation of H5 AIVs from the Eurasian and the American H5 AIVs.
Subject(s)
Antibodies, Monoclonal/chemistry , Enzyme-Linked Immunosorbent Assay/veterinary , Hemagglutinin Glycoproteins, Influenza Virus/analysis , Influenza A Virus, H5N2 Subtype/isolation & purification , Influenza in Birds/virology , Animals , Antibodies, Monoclonal/immunology , Chick Embryo , Chickens , Female , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Hemagglutinin Glycoproteins, Influenza Virus/immunology , Influenza A Virus, H5N2 Subtype/genetics , Influenza A Virus, H5N2 Subtype/immunology , Influenza in Birds/diagnosis , Influenza in Birds/immunology , Limit of Detection , Specific Pathogen-Free OrganismsABSTRACT
The H6 subtype of avian influenza virus (AIV) infection occurs frequently in wild and domestic birds. AIV antigen detection is preferred for controlling AIV as birds are infected before they produce antibodies. The purpose of this study was to develop an early diagnostic method for AIV detection. Six monoclonal antibodies (mAbs) developed from a field H6N1 AIV strain were tested for their ability to bind to viruses. The two that showed the greatest binding ability to AIVs were used for antigen detection. An antigen-capture enzyme-linked immunosorbent assay (ELISA) to detect H6 AIVs was developed using these mAbs. One mAb was coated onto an ELISA plate as the capture antibody. The other mAb was used as the detector antibody after labeling with horseradish peroxidase. The antigen-capture ELISA detected H6N1 AIVs but not H5 AIVs, human H1N1, H3N2 influenza or other viruses. This antigen-capture ELISA could be used to specifically detect H6N1 AIV.