Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5517015 | Biologicals | 2016 | 8 Pages |
â¢Construction of hydrophobic domains deletion protein of NS3 of bluetongue virus.â¢Production of recombinant NS3ÎHD fusion protein from Escherichia coli.â¢Immuno-reactivity of rNS3ÎHD fusion protein to detect BTV-NS3 specific antibodies.â¢Diagnostic sensitivity and specificity of rNS3ÎHD based indirect-ELISA for bluetongue.
Serological diagnostics for bluetongue (BT), which is an infectious, non-contagious and arthropod-borne virus disease of ruminants, are primarily dependent on availability of high quality native or recombinant antigen(s) based on either structural/non-structural proteins in sufficient quantity. Non-structural proteins (NS1-NS4) of BT virus are presumed candidate antigens in development of DIVA diagnostics. In the present study, NS3 fusion gene encoding for NS3 protein containing the N- and C-termini with a deletion of two hydrophobic domains (118A to S141 aa and 162S to A182 aa) and intervening variable central domain (142D to K161 aa) of bluetongue virus 23 was constructed, cloned and over-expressed using prokaryotic expression system. The recombinant NS3ÎHD fusion protein (â¼38 kDa) including hexa-histidine tag on its both termini was found to be non-cytotoxic to recombinant Escherichia coli cells and purified by affinity chromatography. The purified rNS3ÎHD fusion protein was found to efficiently detect BTV-NS3 specific antibodies in indirect-ELISA format with diagnostic sensitivity (DSn = 94.4%) and specificity (DSp = 93.9%). The study indicated the potential utility of rNS3ÎHD fusion protein as candidate diagnostic reagent in developing an indirect-ELISA for sero-surveillance of animals for BTV antibodies under DIVA strategy, wherever monovalent/polyvalent killed BT vaccine formulations devoid of NS proteins are being practiced for immunization.