title: Expression of recombinant small hydrophobic protein for serospecific detection of avian pneumovirus subgroup C.
abstract: The small hydrophobic (SH) gene of the avian pneumovirus (APV) Colorado isolate (CO), which belongs to subgroup C (APV/C), was expressed with a baculovirus vector. The recombinant SH protein was evaluated as a potential subgroup-specific diagnostic reagent in order to differentiate infections resulting from APV/C from those induced by APV/A, APV/B, and human metapneumovirus (hMPV). When the recombinant baculovirus was used to infect insect cells, a 31- to 38-kDa glycosylated form of the SH protein was produced and subsequently tested for reactivity with antibodies specific for APV/A, APV/B, APV/C, and hMPV. Western blot analysis showed that the expressed recombinant SH protein could only be recognized by APV/C-specific antibodies. This result was consistent with sequence analysis of the APV/C SH protein, which had very low (24%) amino acid identity with the corresponding protein of hMPV and no discernible identity with the SH protein of APV/A or APV/B. A recombinant SH protein-based enzyme-linked immunosorbent assay (ELISA) was developed, and it further confirmed the lack of reactivity of this protein with antisera raised to APV/A, APV/B, and hMPV and supported its designation as a subgroup-specific antigen. This finding indicated that the recombinant SH protein was a suitable antigen for ELISA-based detection of subgroup-specific antibodies in turkeys and could be used for serologically based differential diagnosis of APV and hMPV infections.
authors.affiliation: Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Winnipeg, Manitoba R3E 3M4, Canada. luol@inspection.gc.ca
authors.forename: Lizhong
authors.initials: L
authors.lastname: Luo
authors.forename: Marta I
authors.initials: MI
authors.lastname: Sabara
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D014644:Genetic Variation; D006801:Humans; D029121:Metapneumovirus / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D018184:Paramyxoviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary; D011201:Poultry Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015682:Retroviridae Proteins, Oncogenic / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D012698:Serologic Tests; D014422:Turkeys / Q000821:virology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; C056150:small hydrophobic protein, virus