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Development of single-chain Fv against the nucleoprotein of type A influenza virus and its use in ELISA

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Title Development of single-chain Fv against the nucleoprotein of type A influenza virus and its use in ELISA
Not Available
 
Creator Sengupta D.
Shaikh A.
Bhatia S.
Pateriya A.K.
Khandia R.
Sood R.
Prakash A.
Pattnaik B.
Pradhan H.K.
 
Subject AIV
ELISA
Nucleoprotein
Recombinant antibody
ScFv
 
Description Not Available
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.
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Date 2018-03-22T07:48:59Z
2018-03-22T07:48:59Z
2014-11-01
 
Type Research Paper
 
Identifier 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.
0166-0934
http://krishi.icar.gov.in/jspui/handle/123456789/5976
 
Language English
 
Relation Not Available;
 
Publisher Elsevier