Development of monoclonal antibody-linked ELISA for sero-diagnosis of vesicular stomatitis virus (VSV-IN) using baculovirus expressed glycoprotein.
The gene encoding the envelope glycoprotein (GP) of vesicular stomatitis virus serotype, Indiana (VSV-IN), was expressed under the polyhedron promoter of baculovirus. The recombinant GP was applied as a diagnostic antigen for the detection of cattle and horse antibodies to VSV. In addition, the neutralizing monoclonal antibody (Mab) to GP of VSV-IN was used as trapping antibody in a Mab-linked indirect ELISA (MLI-ELISA) or detecting antibody in a Mab-linked competitive ELISA (MLC-ELISA). The diagnostic efficiencies of MLI-ELISA and MLC-ELISA were evaluated with currently available C-ELISA from OIE reference laboratory for vesicular stomatitis as a gold standard by using VSV-positive equine sera and negative bovine sera vaccinated against foot-and-mouth disease (FMD) in the field. When naturally infected equine sera and FMDV vaccinated bovine sera were tested, MLI-ELISA and MLC-ELISA showed relative sensitivities of 80% and 95% with relative specificity of 97% and 99%, respectively. However, both ELISAs cross-reacted with equine sera against New Jersey (VSV-NJ) serotype. The comparison of the two ELISAs revealed that MLC-ELISA was relatively more sensitive and specific than MLI-ELISA, indicating that MLC-ELISA can be applied to sero-diagnosis for VSV-IN infection.
Kweon
Chang Hee
CH
Animal Disease Diagnosis Division, National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry, 480 Anyang, 6 Dong, Gyunggi Do, Republic of Korea. kweonch@nvrqs.go.kr
Kwon
Byung Joon
BJ
Kim
In Joong
IJ
Lee
Se Young
SY
Ko
Young Joon
YJ
D000818:Animals; D000911:Antibodies, Monoclonal; D016367:Baculoviridae / Q000378:metabolism; D002417:Cattle; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D006736:Horses; D011994:Recombinant Proteins / Q000096:biosynthesis; D018353:Rhabdoviridae Infections / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods / Q000662:veterinary*; D014721:Vesicular stomatitis Indiana virus / Q000276:immunology*; D018116:Vesiculovirus; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins