Development and application of an indirect ELISA for the detection of antibodies against encephalomyocarditis virus.

Abstract

Encephalomyocarditis virus (EMCV) can cause acute myocarditis in young pigs or reproductive failure in sows. It has been recognized worldwide as a pathogen infecting many species and causes substantial economic losses. In the present study, an indirect ELISA was developed for the detection of antibodies to EMCV. The VP1 gene of EMCV was amplified by reverse transcription-quantitative polymerase chain reaction and expressed in Escherichia coli with 49.3 kDa under the condition of isopropyl-β-d-thiogalactoside. Following this, the authors obtained the recombinant protein VP1 as a coating antigen. The antigen concentration and serum dilution were optimized using a checkerboard titration. Compared with viral neutralization tests, the sensitivity and specificity of the indirect ELISA was 95.7% and 92.9%, respectively. A total of 265 clinical swine serum samples from different pig farms in China were used to a serological survey. The seropositive rate of the serum samples was 81.9%. In conclusion, the developed indirect ELISA assay is sensitive and specific, which will be useful for large-scale serological survey in EMCV infection and monitoring antibodies titers against EMCV.