Development of recombinant nucleoprotein-based diagnostic systems for lassa fever

Diagnostic systems for Lassa fever (LF), a viral hemorrhagic fever caused by Lassa virus (LASV), such as enzyme immunoassays for the detection of LASV antibodies and LASV antigens, were developed using the recombinant nucleoprotein (rNP) of LASV (LASV-rNP). The LASV-rNP was expressed in a recombinan...

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Autores principales: Saijo, Masayuki, Georges-Courbot, Marie Claude, Marianneau, Philippe, Romanowski, Víctor, Fukushi, Shuetsu, Mizutani, Tetsuya, Georges, Alain Jean, Kurata, Takeshi, Kurane, Ichiro, Morikawa, Shigeru
Formato: Articulo
Lenguaje:Inglés
Publicado: 2007
Materias:
Bioquímica
Lassa fever
recombinant nucleoprotein
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/83208
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-83208
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Bioquímica
Lassa fever
recombinant nucleoprotein
spellingShingle Bioquímica
Lassa fever
recombinant nucleoprotein
Saijo, Masayuki
Georges-Courbot, Marie Claude
Marianneau, Philippe
Romanowski, Víctor
Fukushi, Shuetsu
Mizutani, Tetsuya
Georges, Alain Jean
Kurata, Takeshi
Kurane, Ichiro
Morikawa, Shigeru
Development of recombinant nucleoprotein-based diagnostic systems for lassa fever
topic_facet Bioquímica
Lassa fever
recombinant nucleoprotein
description Diagnostic systems for Lassa fever (LF), a viral hemorrhagic fever caused by Lassa virus (LASV), such as enzyme immunoassays for the detection of LASV antibodies and LASV antigens, were developed using the recombinant nucleoprotein (rNP) of LASV (LASV-rNP). The LASV-rNP was expressed in a recombinant baculovirus system. LASV-rNP was used as an antigen in the detection of LASV-antibodies and as an immunogen for the production of monoclonal antibodies. The LASV-rNP was also expressed in HeLa cells by transfection with the expression vector encoding cDNA of the LASV-NP gene. An immunoglobulin G enzyme-linked immunosorbent assay (ELISA) using LASV-rNP and an indirect immunofluorescence assay using LASV-rNP-expressing HeLa cells were confirmed to have high sensitivity and specificity in the detection of LASV-antibodies. A novel monoclonal antibody to LASV-rNP, monoclonal antibody 4A5, was established. A sandwich antigen capture (Ag-capture) ELISA using the monoclonal antibody and an anti-LASV-rNP rabbit serum as capture and detection antibodies, respectively, was then developed. Authentic LASV nucleoprotein in serum samples collected from hamsters experimentally infected with LASV was detected by the Ag-capture ELISA. The Ag-capture ELISA specifically detected LASV-rNP but not the rNPs of lymphocytic choriomeningitis virus or Junin virus. The sensitivity of the Ag-capture ELISA in detecting LASV antigens was comparable to that of reverse transcription-PCR in detecting LASV RNA. These LASV rNP-based diagnostics were confirmed to be useful in the diagnosis of LF even in institutes without a high containment laboratory, since the antigens can be prepared without manipulation of the infectious viruses.
format Articulo
Articulo
author Saijo, Masayuki
Georges-Courbot, Marie Claude
Marianneau, Philippe
Romanowski, Víctor
Fukushi, Shuetsu
Mizutani, Tetsuya
Georges, Alain Jean
Kurata, Takeshi
Kurane, Ichiro
Morikawa, Shigeru
author_facet Saijo, Masayuki
Georges-Courbot, Marie Claude
Marianneau, Philippe
Romanowski, Víctor
Fukushi, Shuetsu
Mizutani, Tetsuya
Georges, Alain Jean
Kurata, Takeshi
Kurane, Ichiro
Morikawa, Shigeru
author_sort Saijo, Masayuki
title Development of recombinant nucleoprotein-based diagnostic systems for lassa fever
title_short Development of recombinant nucleoprotein-based diagnostic systems for lassa fever
title_full Development of recombinant nucleoprotein-based diagnostic systems for lassa fever
title_fullStr Development of recombinant nucleoprotein-based diagnostic systems for lassa fever
title_full_unstemmed Development of recombinant nucleoprotein-based diagnostic systems for lassa fever
title_sort development of recombinant nucleoprotein-based diagnostic systems for lassa fever
publishDate 2007
url http://sedici.unlp.edu.ar/handle/10915/83208
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