tetano
Editor, Senior Moderator
J Immunoassay Immunochem. 2016 Feb 1. [Epub ahead of print]
[h=1]EXPRESSION AND PURIFICATION OF RECOMBINANT H5HA1 PROTEIN OF H5N1 AVIAN INFLUENZA VIRUS IN E. coli AND ITS APPLICATION IN INDIRECT ELISA.[/h] Redda YT[SUP]1[/SUP], Venkatesh G[SUP]1[/SUP], Kalaiyarasu S[SUP]1[/SUP], Bhatia S[SUP]1[/SUP], Senthil Kumar D[SUP]1[/SUP], Nagarajan S[SUP]1[/SUP], Pillai A[SUP]1[/SUP], Tripathi S[SUP]1[/SUP], Kulkarni DD[SUP]1[/SUP], Dubey SC[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] The PCR amplified HA1 fragment of H5N1 (H5HA1) avian influenza virus (AIV) haemagglutinin gene was cloned into pET28a (+) expression vector and expressed in Rosetta Blue (DE3) pLysS cells. The recombinant H5HA1 (rH5HA1) protein purified by passive gel elution after SDS-PAGE of the inclusion bodies reacted specifically with H5N1 serum in Western blot analysis. A subtype specific indirect enzyme linked immunosorbent assay (iELISA) using the rH5HA1 protein as the coating antigen was developed for detecting antibodies to H5 subtype of AIV. The assay had 89.04% sensitivity and 95.95% specificity when compared with haemagglutination inhibition test. The Kappa value of 0.842 indicated a perfect agreement between the tests. The iELISA developed can be used for serosurveillance of avian influenza in chickens.
[h=4]KEYWORDS:[/h] AIV H5N1; characterization; expression; haemagglutinin; indirect ELISA
PMID: 26829111 [PubMed - as supplied by publisher]
[h=1]EXPRESSION AND PURIFICATION OF RECOMBINANT H5HA1 PROTEIN OF H5N1 AVIAN INFLUENZA VIRUS IN E. coli AND ITS APPLICATION IN INDIRECT ELISA.[/h] Redda YT[SUP]1[/SUP], Venkatesh G[SUP]1[/SUP], Kalaiyarasu S[SUP]1[/SUP], Bhatia S[SUP]1[/SUP], Senthil Kumar D[SUP]1[/SUP], Nagarajan S[SUP]1[/SUP], Pillai A[SUP]1[/SUP], Tripathi S[SUP]1[/SUP], Kulkarni DD[SUP]1[/SUP], Dubey SC[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] The PCR amplified HA1 fragment of H5N1 (H5HA1) avian influenza virus (AIV) haemagglutinin gene was cloned into pET28a (+) expression vector and expressed in Rosetta Blue (DE3) pLysS cells. The recombinant H5HA1 (rH5HA1) protein purified by passive gel elution after SDS-PAGE of the inclusion bodies reacted specifically with H5N1 serum in Western blot analysis. A subtype specific indirect enzyme linked immunosorbent assay (iELISA) using the rH5HA1 protein as the coating antigen was developed for detecting antibodies to H5 subtype of AIV. The assay had 89.04% sensitivity and 95.95% specificity when compared with haemagglutination inhibition test. The Kappa value of 0.842 indicated a perfect agreement between the tests. The iELISA developed can be used for serosurveillance of avian influenza in chickens.
[h=4]KEYWORDS:[/h] AIV H5N1; characterization; expression; haemagglutinin; indirect ELISA
PMID: 26829111 [PubMed - as supplied by publisher]