Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae

West Nile virus (WNV), a Flavivirus distributed most widely, is presenting lately variable epidemiological and ecological patterns, including an increasing virulence that has already caused over 1000 human deaths in USA. Currently, diagnosis of WNV is achieved mainly by enzyme-linked immunoassays (ELISAs) based on the use of inactivated whole WNV (iWNV) as antigen, although results have to be confirmed by plaque reduction neutralization tests (PRNTs). Expression of WNV envelope recombinant E (rE) protein and its usefulness as ELISA antigen are described. Production of rE was achieved upon infection of Trichoplusia ni insect larvae with a recombinant baculovirus. Once optimized, the rE-based ELISA was validated with a battery of mouse and equine sera characterized previously. Concordance with the iWNV-based ELISA used routinely was good (95%), as it was with the reference PRNT (90%), with specificity of 94.4% and sensitivity of 88.1%. Production of rE protein in insect larvae allows for an easy, low cost and quite large-scale yield of partially purified antigen which is suitable for serological diagnosis of WNV, without the need for manipulation of large quantities of infective virus. © 2010 Elsevier B.V.

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Main Authors: Alonso-Padilla, J., Jiménez De Oya, Nereida, Blázquez Martín, Ana Belén, Loza-Rubio, E., Escribano Romero, Estela, Saiz Calahorra, Juan Carlos
Format: artículo biblioteca
Language:English
Published: Elsevier 2010
Subjects:WNV, E protein, Baculovirus, Larvae, ELISA,
Online Access:http://hdl.handle.net/20.500.12792/6048
http://hdl.handle.net/10261/290761
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spelling dig-inia-es-10261-2907612023-02-17T12:29:07Z Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae Alonso-Padilla, J. Jiménez De Oya, Nereida Blázquez Martín, Ana Belén Loza-Rubio, E. Escribano Romero, Estela Saiz Calahorra, Juan Carlos Escribano Romero, Estela WNV E protein Baculovirus Larvae ELISA West Nile virus (WNV), a Flavivirus distributed most widely, is presenting lately variable epidemiological and ecological patterns, including an increasing virulence that has already caused over 1000 human deaths in USA. Currently, diagnosis of WNV is achieved mainly by enzyme-linked immunoassays (ELISAs) based on the use of inactivated whole WNV (iWNV) as antigen, although results have to be confirmed by plaque reduction neutralization tests (PRNTs). Expression of WNV envelope recombinant E (rE) protein and its usefulness as ELISA antigen are described. Production of rE was achieved upon infection of Trichoplusia ni insect larvae with a recombinant baculovirus. Once optimized, the rE-based ELISA was validated with a battery of mouse and equine sera characterized previously. Concordance with the iWNV-based ELISA used routinely was good (95%), as it was with the reference PRNT (90%), with specificity of 94.4% and sensitivity of 88.1%. Production of rE protein in insect larvae allows for an easy, low cost and quite large-scale yield of partially purified antigen which is suitable for serological diagnosis of WNV, without the need for manipulation of large quantities of infective virus. © 2010 Elsevier B.V. 2023-02-17T12:29:07Z 2023-02-17T12:29:07Z 2010 artículo Journal of Virological Methods 166: 37-41 (2010) 0166-0934 http://hdl.handle.net/20.500.12792/6048 http://hdl.handle.net/10261/290761 10.1016/j.jviromet.2010.02.013 en none Elsevier
institution INIA ES
collection DSpace
country España
countrycode ES
component Bibliográfico
access En linea
databasecode dig-inia-es
tag biblioteca
region Europa del Sur
libraryname Biblioteca del INIA España
language English
topic WNV
E protein
Baculovirus
Larvae
ELISA
WNV
E protein
Baculovirus
Larvae
ELISA
spellingShingle WNV
E protein
Baculovirus
Larvae
ELISA
WNV
E protein
Baculovirus
Larvae
ELISA
Alonso-Padilla, J.
Jiménez De Oya, Nereida
Blázquez Martín, Ana Belén
Loza-Rubio, E.
Escribano Romero, Estela
Saiz Calahorra, Juan Carlos
Escribano Romero, Estela
Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae
description West Nile virus (WNV), a Flavivirus distributed most widely, is presenting lately variable epidemiological and ecological patterns, including an increasing virulence that has already caused over 1000 human deaths in USA. Currently, diagnosis of WNV is achieved mainly by enzyme-linked immunoassays (ELISAs) based on the use of inactivated whole WNV (iWNV) as antigen, although results have to be confirmed by plaque reduction neutralization tests (PRNTs). Expression of WNV envelope recombinant E (rE) protein and its usefulness as ELISA antigen are described. Production of rE was achieved upon infection of Trichoplusia ni insect larvae with a recombinant baculovirus. Once optimized, the rE-based ELISA was validated with a battery of mouse and equine sera characterized previously. Concordance with the iWNV-based ELISA used routinely was good (95%), as it was with the reference PRNT (90%), with specificity of 94.4% and sensitivity of 88.1%. Production of rE protein in insect larvae allows for an easy, low cost and quite large-scale yield of partially purified antigen which is suitable for serological diagnosis of WNV, without the need for manipulation of large quantities of infective virus. © 2010 Elsevier B.V.
format artículo
topic_facet WNV
E protein
Baculovirus
Larvae
ELISA
author Alonso-Padilla, J.
Jiménez De Oya, Nereida
Blázquez Martín, Ana Belén
Loza-Rubio, E.
Escribano Romero, Estela
Saiz Calahorra, Juan Carlos
Escribano Romero, Estela
author_facet Alonso-Padilla, J.
Jiménez De Oya, Nereida
Blázquez Martín, Ana Belén
Loza-Rubio, E.
Escribano Romero, Estela
Saiz Calahorra, Juan Carlos
Escribano Romero, Estela
author_sort Alonso-Padilla, J.
title Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae
title_short Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae
title_full Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae
title_fullStr Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae
title_full_unstemmed Evaluation of an enzyme-linked immunosorbent assay for detection of West Nile virus infection based on a recombinant envelope protein produced in Trichoplusia ni larvae
title_sort evaluation of an enzyme-linked immunosorbent assay for detection of west nile virus infection based on a recombinant envelope protein produced in trichoplusia ni larvae
publisher Elsevier
publishDate 2010
url http://hdl.handle.net/20.500.12792/6048
http://hdl.handle.net/10261/290761
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