title: Use of a Japanese quail fibrosarcoma cell line (QT-35) in serologic assays to determine the antigenic relationship of avian metapneumoviruses.
abstract: The ability of a Japanese quail fibrosarcoma cell line (QT-35) to support the replication of avian metapneumoviruses belonging to the 3 subgroups A (14/1 virus), B (Colorado virus), and C (Hungary virus) enabled the development of assays for the detection and evaluation of virus-specific antibodies. On the basis of the results of enzyme-linked immunosorbent assay (ELISA), plaque reduction neutralization assay (PRNA), immunofluorescent assay (IFA), and Western blot analysis, some degree of antigenic cross-reactivity was observed between prototype viruses belonging to each of the 3 subgroups A, B, and C. The antigen produced in QT-35 cells was found to be superior with respect to its reactivity with virus-specific antibodies, as determined when used in ELISA and IFA. Standardization of both the input virus and the virus-specific antibodies in PRNA enabled a more detailed analysis of the antigenic relationship between these viruses. Specifically, it was observed that 14/1 virus shared more neutralizing regions with Hungary and Colorado viruses than did either of these viruses with 14/1 virus. In addition, Hungary virus shared comparatively fewer neutralizing epitopes with the Colorado virus than did 14/1 virus. Western blot analysis of the reactivity patterns of virus antigen, produced in QT-35 cells, with subgroup-specific antibodies identified a cross-reactive protein migrating at approximately 18 kD. These assays and the information from the Western blot will enable further analysis of avian metapneumovirus isolates to determine antigenic relationships.
authors.affiliation: Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, Manitoba R3E 3M4, Canada.
authors.forename: Marta I
authors.initials: MI
authors.lastname: Sabara
authors.forename: June E
authors.initials: JE
authors.lastname: Larence
authors.forename: Stacey
authors.initials: S
authors.lastname: Halayko
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D002522:Chlorocebus aethiops; D003370:Coturnix; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005354:Fibrosarcoma / Q000276:immunology* / Q000662:veterinary*; D029121:Metapneumovirus / Q000145:classification / Q000276:immunology*; D014407:Tumor Cells, Cultured; D014709:Vero Cells
chemical_list: D000956:Antigens, Viral