Enhanced expression of the Erns protein of classical swine fever virus in yeast and its application in an indirect enzyme-linked immunosorbent assay for antibody differentiation of infected from vaccinated animals
Enhanced expression of the Erns protein of classical swine fever virusin yeast and its application in an indirect enzyme-linkedimmunosorbent assay for antibody differentiation of infected fromvaccinated animals.pdf
[en] tClassical swine fever (CSF), caused by classical swine fever virus (CSFV), is a devastating disease of swineworldwide. Although a mandatory vaccination with the modified live vaccine C-strain has been imple-mented in China for decades, CSF remains a serious threat to the swine industry. To facilitate the controland eradication of CSF in China, the E2-based marker vaccine rAdV-SFV-E2, an adenovirus-delivered,alphavirus replicon-vectored vaccine, has been developed. Accordingly, an accompanying discriminatorytest that allows differentiating infected from vaccinated animals (DIVA) is required. Here, the enhancedexpression of Ernsprotein of CSFV was achieved in the methyltropic yeast Pichia pastoris by codon-optimization of the Ernsgene, and an indirect enzyme-linked immunosorbent assay (iELISA) based on theyeast-expressed Erns(yErns) was developed and evaluated. The optimized iELISA was able to detect CSFV-specific antibodies in the serum samples from the CSFV-infected pigs as early as 6 days post-infection,and discriminate the CSFV-infected pigs from those vaccinated with rAdV-SFV-E2. The iELISA was evalu-ated using a panel of swine sera, and showed comparable sensitivity (94.6%) and specificity (97.1%), andthe consistence rates with the virus neutralization test were 96.8% for CSFV-infected swine sera, 83.3%for C-strain-vaccinated swine sera, and 95.0% for field swine sera. In addition, the iELISA showed highersensitivity (90.4%) compared with PrioCHECK CSFV Erns(59.6%). Taken together, the yErns-based iELISA isspecific and sensitive, representing a promising DIVA test for E2-based marker vaccines against CSF.
Research center :
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Luo, Yuzi ✱
Li, Lin ✱; Université de Liège - ULiège > Doct. sc. agro. & ingé. biol. (Paysage)
✱ These authors have contributed equally to this work.
Enhanced expression of the Erns protein of classical swine fever virus in yeast and its application in an indirect enzyme-linked immunosorbent assay for antibody differentiation of infected from vaccinated animals
Publication date :
21 May 2015
Journal title :
Journal of Virological Methods
ISSN :
0166-0934
eISSN :
1879-0984
Publisher :
Elsevier, Netherlands
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
Development of Livestock Genetic Engineering Vaccine
Funders :
National 863Projects of China (No. 2011AA10A208) and the European Union’sSeventh Framework Programme LinkTADs (No. 613804).
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