Serological detection of bovine ephemeral fever virus using an indirect ELISA based on antigenic site G1 expressed in Pichia pastoris.
An indirect ELISA for the serological detection of bovine ephemeral fever virus (BEFV) infection in cattle is described in which a glycosylated protein of approximately 25 kDa (including the G1 antigenic site of the virus glycoprotein) expressed in Pichia pastoris GS115 was used as the coating antigen. The optimal concentration of coated antigen was 0.3 microg/well at a serum dilution of 1:40. The optimal positive threshold value of the assay was 1.88, as derived from receiver operating characteristic curve analysis. The test had 100% sensitivity and 96.7% specificity when compared with a micro-neutralisation test using 336 positive and 180 negative sera to BEFV, respectively. The inter-assay and intra-assay coefficients of variation for 15 sera were both <5.8% and there was no evidence of cross-reactivity between the tested coating antigen and antibodies to related rabies virus. The ELISA is an inexpensive and rapid serological detection method that would be suitable for screening for BEFV infection on a large scale.
Zheng
Fu-Ying
FY
Key Laboratories of Grazing Animal Diseases and Animal Virology, Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu 730046, China.
Lin
Guo-Zhen
GZ
Qiu
Chang-Qing
CQ
Zhou
Ji-Zhang
JZ
Cao
Xiao-An
XA
Gong
Xiao-Wei
XW
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards / Q000662:veterinary*; D004810:Ephemeral Fever / Q000175:diagnosis*; D018115:Ephemeral Fever Virus, Bovine / Q000276:immunology*; D010843:Pichia / Q000276:immunology; D012372:ROC Curve; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D000956:Antigens, Viral