title: Production and characterization of single-chain antibody (scFv) against 3ABC non-structural protein in Escherichia coli for sero-diagnosis of Foot and Mouth Disease virus.
abstract: Differentiation of Foot-and-Mouth Disease infected from vaccinated animals is essential for effective implementation of vaccination based control programme. Detection of antibodies against 3ABC non-structural protein of FMD virus by immunodiagnostic assays provides reliable indication of FMD infection. Sero-monitoring of FMD in the large country like India is a big task where thousands of serum samples are annually screened. Currently, monoclonal or polyclonal antibodies are widely used in these immunodiagnostic assays. Considering the large population of livestock in the country, an economical and replenishable alternative of these antibodies was required. In this study, specific short chain variable fragment (scFv) antibody against 3B region of 3ABC poly-protein was developed. High level of scFv expression in Escherichia coli system was obtained by careful optimization in four different strains. Two formats of enzyme immunoassays (sandwich and competitive ELISAs) were optimized using scFv with objective to differentiate FMD infected among the vaccinated population. The assays were statistically validated by testing 2150 serum samples. Diagnostic sensitivity/specificity of sandwich and competitive ELISAs were determined by ROC method as 92.2%/95.5% and 89.5%/93.5%, respectively. This study demonstrated that scFv is a suitable alternate for immunodiagnosis of FMD on large scale.
authors.affiliation: Project Directorate on Foot and Mouth Disease, Indian Council of Agricultural Research, IVRI Campus, Mukteswar, Uttarakhand 263138, India.
authors.forename: Gaurav K
authors.initials: GK
authors.lastname: Sharma
authors.affiliation: Project Directorate on Foot and Mouth Disease, Indian Council of Agricultural Research, IVRI Campus, Mukteswar, Uttarakhand 263138, India.
authors.forename: Sonalika
authors.initials: S
authors.lastname: Mahajan
authors.affiliation: Project Directorate on Foot and Mouth Disease, Indian Council of Agricultural Research, IVRI Campus, Mukteswar, Uttarakhand 263138, India.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Matura
authors.affiliation: Project Directorate on Foot and Mouth Disease, Indian Council of Agricultural Research, IVRI Campus, Mukteswar, Uttarakhand 263138, India.
authors.forename: Saravanan
authors.initials: S
authors.lastname: Subramaniam
authors.affiliation: Project Directorate on Foot and Mouth Disease, Indian Council of Agricultural Research, IVRI Campus, Mukteswar, Uttarakhand 263138, India.
authors.forename: Jajati K
authors.initials: JK
authors.lastname: Mohapatra
authors.affiliation: Project Directorate on Foot and Mouth Disease, Indian Council of Agricultural Research, IVRI Campus, Mukteswar, Uttarakhand 263138, India. Electronic address: pattnaikb@gmail.com.
authors.forename: Bramhadev
authors.initials: B
authors.lastname: Pattnaik
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000914:Antibodies, Viral / Q000097:blood; D002020:Buffaloes; D002417:Cattle; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism*; D005536:Foot-and-Mouth Disease / Q000175:diagnosis*; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D006825:Hybridomas / Q000378:metabolism; D007124:Immunoenzyme Techniques; D051379:Mice; D009997:Osmotic Pressure; D012372:ROC Curve; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D057127:Single-Chain Antibodies / Q000737:chemistry*; D014765:Viral Vaccines
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014765:Viral Vaccines
keywords: 3ABC; DIVA; Foot-and-mouth disease; ROC; scFv