Expression of recombinant small hydrophobic protein for serospecific detection of avian pneumovirus subgroup C.
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.
Luo
Lizhong
L
Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Winnipeg, Manitoba R3E 3M4, Canada. luol@inspection.gc.ca
Sabara
Marta I
MI
Li
Yan
Y
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
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; C056150:small hydrophobic protein, virus