An indirect ELISA of classical swine fever virus based on quadruple antigenic epitope peptide expressed in E.coli.
In this study, a synthesized quadruple antigenic epitope gene region of the classical swine fever virus (CSFV) E2 glycoprotein was expressed in E. coli to a obtain target protein. This target protein was used as a coating antigen to establish an indirect ELISA for specifically detecting anti-CSFV antibodies in serum samples from pigs. The P/N cut-off value of this assay was 1.92 by receiver operating characteristic curve (ROC) analysis based on 30 negative sera and 80 positive samples. The test gave 97.5% sensitivity and 96.7% specificity compared with the indirect hemagglutination (IHA) test. The inter-assay and intra-assay coefficients of variation (CVs) for 16 sera were both ≤6.8%. No cross-reactivity between the coating antigen and anti-bovine viral diarrhoea virus (BVDV) antibodies was observed.
Key Laboratories of Grazing Animal Diseases and Animal Virology, Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China.
Guo-zhen
GZ
Lin
Fu-ying
FY
Zheng
Ji-zhang
JZ
Zhou
Xiao-an
XA
Cao
Xiao-wei
XW
Gong
Guang-hua
GH
Wang
Chang-qing
CQ
Qiu
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D006691:Classical Swine Fever / Q000175:diagnosis*; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D012372:ROC Curve; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology; D014773:Virology / Q000379:methods*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus