Establishment and application of a solid-phase blocking ELISA method for detection of antibodies against classical swine fever virus

Abstract

Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry.

Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples.

Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test.

Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies.

Conclusions: The spbELISA has practical applications in assessing the vaccination status of large pig herds.

Keywords: Classical swine fever virus; E2 protein; antibody detection; blocking ELISA.