title: Chimeric tymovirus-like particles displaying foot-and-mouth disease virus non-structural protein epitopes and its use for detection of FMDV-NSP antibodies.
abstract: Expression of Physalis mottle tymovirus (PhMV) coat protein (CP) in Escherichia coli (E. coli) was earlier shown to self-assemble into empty capsids that are nearly identical to the capsids formed in vivo. Aminoacid substitutions were made at the N-terminus of wild-type PhMV CP with single or tandem repeats of infection related B-cell epitopes of foot-and-mouth disease virus (FMDV) non-structural proteins (NSPs) 3B1, 3B2, 3AB, 3D and 3ABD of lengths 48, 66, 49, 51 and 55, respectively to produce chimeras pR-Ph-3B1, pR-Ph-3B2, pR-Ph- 3AB, pR-Ph-3D and pR-Ph-3ABD. Expression of these constructs in E. coli resulted in chimeric proteins which self-assembled into chimeric tymovirus-like particles (TVLPs), Ph-3B1, Ph-3B2, Ph-3AB, Ph-3D and Ph-3ABD as determined by ultracentrifugation and electron microscopy. Ph-3B1, Ph-3B2, Ph-3AB and Ph-3ABD reacted with polyclonal anti-3AB antibodies in ELISA and electroblot immunoassay, while wild-type PhMV TVLP and Ph-3D antigens did not react. An indirect ELISA (I-ELISA) was developed using Ph-3AB to detect FMDV-NSP antibodies in sera of animals that showed clinical signs of FMD. Field serum samples from cattle, buffalos, sheep, goats and pigs were examined by using these chimeric TVLPs for the differentiation of FMDV infected animals from vaccinated animals (DIVA). The assay was demonstrated to be highly specific (100%) and reproducible with sensitivity levels (94%) comparable to the Ceditest kit (P>0.05).
authors.affiliation: Indian Immunologicals Limited, Rakshapuram, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: Masarapu
authors.initials: M
authors.lastname: Hema
authors.forename: Singanallur Balasubramanian
authors.initials: SB
authors.lastname: Nagendrakumar
authors.forename: Reddivari
authors.initials: R
authors.lastname: Yamini
authors.forename: Dev
authors.initials: D
authors.lastname: Chandran
authors.forename: Lingala
authors.initials: L
authors.lastname: Rajendra
authors.forename: Dorairajan
authors.initials: D
authors.lastname: Thiagarajan
authors.forename: Satya
authors.initials: S
authors.lastname: Parida
authors.forename: David James
authors.initials: DJ
authors.lastname: Paton
authors.forename: Villuppanoor Alwar
authors.initials: VA
authors.lastname: Srinivasan
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology; D002020:Buffaloes; D036022:Capsid Proteins / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli / Q000378:metabolism; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D006041:Goats; D006801:Humans; D007118:Immunoassay; D008854:Microscopy, Electron; D050506:Mutant Chimeric Proteins / Q000276:immunology; D012756:Sheep; D017806:Tymovirus / Q000235:genetics* / Q000276:immunology* / Q000648:ultrastructure; D017361:Viral Nonstructural Proteins / Q000032:analysis / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D000939:Epitopes; D050506:Mutant Chimeric Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins