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Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli

BACKGROUND: Antigens for Hantavirus serological tests have been produced using DNA recombinant technology for more than twenty years. Several different strategies have been used for that purpose. All of them avoid the risks and difficulties involved in multiplying Hantavirus in the laboratory. In Br...

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Autores principales: Machado, Alex M, Machado, Aline RSR, Moreli, Marcos L, Ribeiro, Bergmann M, Figueiredo, Luiz TM, Wolff, Jose LC
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3114775/
https://www.ncbi.nlm.nih.gov/pubmed/21569341
http://dx.doi.org/10.1186/1743-422X-8-218
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author Machado, Alex M
Machado, Aline RSR
Moreli, Marcos L
Ribeiro, Bergmann M
Figueiredo, Luiz TM
Wolff, Jose LC
author_facet Machado, Alex M
Machado, Aline RSR
Moreli, Marcos L
Ribeiro, Bergmann M
Figueiredo, Luiz TM
Wolff, Jose LC
author_sort Machado, Alex M
collection PubMed
description BACKGROUND: Antigens for Hantavirus serological tests have been produced using DNA recombinant technology for more than twenty years. Several different strategies have been used for that purpose. All of them avoid the risks and difficulties involved in multiplying Hantavirus in the laboratory. In Brazil, the Araraquara virus is one of the main causes of Hantavirus Cardio-Pulmonary Syndrome (HCPS). METHODS: In this investigation, we report the expression of the N protein of the Araraquara Hantavirus in a Baculovirus Expression System, the use of this protein in IgM and IgG ELISA and comparison with the same antigen generated in E. coli. RESULTS: The protein obtained, and purified in a nickel column, was effectively recognized by antibodies from confirmed HCPS patients. Comparison of the baculovirus generated antigen with the N protein produced in E. coli showed that both were equally effective in terms of sensitivity and specificity. CONCLUSIONS: Our results therefore indicate that either of these proteins can be used in serological tests in Brazil.
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spelling pubmed-31147752011-06-15 Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli Machado, Alex M Machado, Aline RSR Moreli, Marcos L Ribeiro, Bergmann M Figueiredo, Luiz TM Wolff, Jose LC Virol J Research BACKGROUND: Antigens for Hantavirus serological tests have been produced using DNA recombinant technology for more than twenty years. Several different strategies have been used for that purpose. All of them avoid the risks and difficulties involved in multiplying Hantavirus in the laboratory. In Brazil, the Araraquara virus is one of the main causes of Hantavirus Cardio-Pulmonary Syndrome (HCPS). METHODS: In this investigation, we report the expression of the N protein of the Araraquara Hantavirus in a Baculovirus Expression System, the use of this protein in IgM and IgG ELISA and comparison with the same antigen generated in E. coli. RESULTS: The protein obtained, and purified in a nickel column, was effectively recognized by antibodies from confirmed HCPS patients. Comparison of the baculovirus generated antigen with the N protein produced in E. coli showed that both were equally effective in terms of sensitivity and specificity. CONCLUSIONS: Our results therefore indicate that either of these proteins can be used in serological tests in Brazil. BioMed Central 2011-05-11 /pmc/articles/PMC3114775/ /pubmed/21569341 http://dx.doi.org/10.1186/1743-422X-8-218 Text en Copyright ©2011 Machado et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Machado, Alex M
Machado, Aline RSR
Moreli, Marcos L
Ribeiro, Bergmann M
Figueiredo, Luiz TM
Wolff, Jose LC
Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli
title Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli
title_full Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli
title_fullStr Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli
title_full_unstemmed Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli
title_short Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli
title_sort expression of recombinant araraquara hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in escherichia coli
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3114775/
https://www.ncbi.nlm.nih.gov/pubmed/21569341
http://dx.doi.org/10.1186/1743-422X-8-218
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