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J Clin Microbiol ; 38(11): 4010-4, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11060061

ABSTRACT

The matrix (M) protein of avian pneumovirus (APV) was evaluated for its antigenicity and reliability in an enzyme-linked immunosorbent assay (ELISA) for diagnosis of APV infection, a newly emergent disease of turkeys in United States. Sera from APV-infected turkeys consistently contained antibodies to a 30-kDa protein (M protein). An ELISA based on recombinant M protein generated in Escherichia coli was compared with the routine APV ELISA that utilizes inactivated virus as antigen. Of 34 experimentally infected turkeys, 33 (97.1%) were positive by M protein ELISA whereas only 18 (52.9%) were positive by routine APV ELISA 28 days after infection. None of the serum samples from 41 uninfected experimental turkeys were positive by M protein ELISA. Of 184 field sera from turkey flocks suspected of having APV infection, 133 (72.3%) were positive by M protein ELISA whereas only 99 (53.8%) were positive by routine APV ELISA. Twelve serum samples, which were negative by M protein ELISA but positive by routine APV ELISA, were not reactive with either recombinant M protein or denatured purified APV proteins by Western analysis. This indicates that the samples had given false-positive results by routine APV ELISA. The M protein ELISA was over six times more sensitive than virus isolation (11.5%) in detecting infections from samples obtained from birds showing clinical signs of APV infection. Taken together, these results show that ELISA based on recombinant M protein is a highly sensitive and specific test for detecting antibodies to APV.


Subject(s)
Antibodies, Viral/blood , Pneumovirus Infections/veterinary , Pneumovirus/immunology , Turkeys , Viral Matrix Proteins/immunology , Animals , Blotting, Western , Enzyme-Linked Immunosorbent Assay/methods , Pneumovirus/isolation & purification , Pneumovirus Infections/diagnosis , Pneumovirus Infections/virology , Poultry Diseases/diagnosis , Poultry Diseases/virology , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and Specificity , Viral Matrix Proteins/genetics
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