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. 1998 Apr;36(4):1105-8.
doi: 10.1128/JCM.36.4.1105-1108.1998.

Baculovirus expression of the fusion protein gene of bovine respiratory syncytial virus and utility of the recombinant protein in a diagnostic enzyme immunoassay

M K Pastey  1 , S K Samal
Affiliations

Baculovirus expression of the fusion protein gene of bovine respiratory syncytial virus and utility of the recombinant protein in a diagnostic enzyme immunoassay

M K Pastey et al. J Clin Microbiol. 1998 Apr.

Abstract

The fusion (F) protein of bovine respiratory syncytial virus (BRSV) was expressed by using a baculovirus vector. Antigenicity was tested by immunofluorescence analysis with F-specific monoclonal and polyclonal antibodies. Antibodies to recombinant F protein raised in a rabbit neutralized BRSV and human respiratory syncytial virus infectivity when tested in a plaque reduction assay. The recombinant F protein was evaluated as a source of antigen in an enzyme-linked immunosorbent assay (ELISA), and this ELISA was compared with the virus neutralization (VN) test for detecting BRSV antibodies in 10 consecutive serum samples from four calves vaccinated with a live modified BRSV vaccine and from two nonvaccinated control calves. The ELISA with the baculovirus-expressed F protein as an antigen compared favorably with the VN test and is a rapid, sensitive, and specific method for detecting serum antibodies to BRSV.

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Figures

FIG. 1
FIG. 1
Expression of F protein by recombinant baculovirus and confirmation of the authenticity of recombinant F protein. Lanes: A, molecular weight standards; B, recombinant baculovirus-infected cell lysate immunoprecipitated with BRSV-specific antiserum; C, BRSV-infected BTu cell lysate immunoprecipitated with BRSV antiserum; D, recombinant baculovirus-infected cell lysate immunoprecipitated with baculovirus-expressed F-specific antiserum; E, recombinant baculovirus-infected cell lysate; F, BRSV-infected BTu cell lysate; G, wild-type AcNPV-infected cell lysate; H, molecular weight standards. Numbers are molecular weights, in thousands.
FIG. 2
FIG. 2
ELISA and VN test results obtained with serum samples from four vaccinated calves. Calves were inoculated intramuscularly on day 0 with an attenuated BRSV vaccine, and a booster vaccination was given after 4 weeks. The ELISA and the VN test were performed as described in the text.
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