title: Indirect-blocking ELISA for detecting antibodies against glycoprotein B (gB) of porcine cytomegalovirus (PCMV).
abstract: The major epitope region of the glycoprotein B (gB) gene of the porcine cytomegalovirus (PCMV), with a length of 270 bp, was cloned and expressed in Escherichia coli Rosetta (DE3). The major gB epitope was detected using an agar gel precipitation and Western blot analysis with the polyclonal antibodies specific for the major epitope. An indirect-blocking enzyme-linked immunosorbent assay (ELISA) was developed using the expressed major gB epitope as a coating antigen for the detection of PCMV antibodies. The results of the tests show that the indirect-blocking ELISA has 98% specificity and 97.8% sensitivity. No cross-reactions were observed between the major gB epitope and the antibodies against other virus, which indicates that the gB epitope is specific for PCMV antibodies. The indirect-blocking ELISA is a highly specific, sensitive method for detecting anti-PCMV gB antibodies.
authors.affiliation: Animal Biotechnology Center, College of Veterinary Medicine of Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.forename: Ling
authors.initials: L
authors.lastname: Zhu
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Shi
authors.forename: Zhiwen
authors.initials: Z
authors.lastname: Xu
authors.forename: Miao
authors.initials: M
authors.lastname: Mei
authors.forename: Weiwei
authors.initials: W
authors.lastname: Xu
authors.forename: Yuancheng
authors.initials: Y
authors.lastname: Zhou
authors.forename: Wanzhu
authors.initials: W
authors.lastname: Guo
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics
chemical_list: D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus