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http://krishi.icar.gov.in/jspui/handle/123456789/5976| Title: | Development of single-chain Fv against the nucleoprotein of type A influenza virus and its use in ELISA |
| Other Titles: | Not Available |
| Authors: | Sengupta D. Shaikh A. Bhatia S. Pateriya A.K. Khandia R. Sood R. Prakash A. Pattnaik B. Pradhan H.K. |
| Published/ Complete Date: | 2014-11-01 |
| Project Code: | Not Available |
| Keywords: | AIV ELISA Nucleoprotein Recombinant antibody ScFv |
| Publisher: | Elsevier |
| Citation: | Sengupta D, Shaikh A, Bhatia S, Pateriya AK, Khandia R, Sood R, Prakash A, Pattnaik B, Pradhan HK(2014), Development of single-chain Fv against the nucleoprotein of type A influenza virus and its use in ELISA.J Virol Methods. 2014 Nov;208:129-37. |
| Series/Report no.: | Not Available; |
| Abstract/Description: | Single chain fragment variable (ScFv) antibodies specific to the nucleoprotein (NP) of avian influenza virus (AIV) were developed using a phage display system. The variable heavy (VH) and the variable light (VL) chain gene fragments were derived from spleen cells of Balb/c mouse immunized with a recombinant NP (rNP) antigen (∼63 kDa) of H5N1 influenza virus. The VH and the VL DNA fragments were assembled through a flexible linker DNA to generate ScFv DNA that was cloned subsequently in a phagemid to express ScFv protein in Escherichia coli cells. The specific reactivity of the ScFv with the rNP antigen and viral antigen (H5N1) was confirmed by Western blot and ELISA. A competitive inhibition ELISA (CI-ELISA) was developed using the rNP and the anti-NP ScFv for detection of type-specific antibodies to AIV in chicken sera. The ScFv based CI-ELISA was compared with hemagglutination inhibition (HI) test and agar gel immunodiffusion (AGID) test over 850 sera. Sensitivity of the CI-ELISA was 100% with HI and AGID and specificity was 98.7% with HI and 100% with AGID. Copyright © 2014 Elsevier B.V. All rights reserved. |
| Description: | Not Available |
| ISSN: | 0166-0934 |
| Type(s) of content: | Research Paper |
| Sponsors: | Not Available |
| Language: | English |
| Name of Journal: | Journal of Virological Method |
| NAAS Rating: | 7.79 |
| Volume No.: | 208 |
| Page Number: | 129-37 |
| Source, DOI or any other URL: | 10.1016/j.jviromet.2014.08.009 |
| URI: | http://krishi.icar.gov.in/jspui/handle/123456789/5976 |
| Appears in Collections: | AS-NIHSAD-Publication |
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