Cargando…

Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens

BACKGROUND: Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis. To detect RVF virus (RVFV) infection, indirect immunoglobulin G (IgG) and immunoglobulin M (IgM) enzyme linked immunosorbent assays (ELISAs) which utilize recombinant RVFV nucleocapsid (RVFV-N) protein as assay antigen, have rep...

Descripción completa

Detalles Bibliográficos
Autores principales: Yu, Fuxun, Adungo, Ferdinard, Konongoi, Samson Limbaso, Inoue, Shingo, Sang, Rosemary, Ashur, Salame, Kwallah, Allan ole, Uchida, Leo, Buerano, Corazon C, Mwau, Matilu, Zha, Yan, Nie, Yingjie, Morita, Kouichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6249750/
https://www.ncbi.nlm.nih.gov/pubmed/30466469
http://dx.doi.org/10.1186/s12985-018-1071-y
_version_ 1783372809467068416
author Yu, Fuxun
Adungo, Ferdinard
Konongoi, Samson Limbaso
Inoue, Shingo
Sang, Rosemary
Ashur, Salame
Kwallah, Allan ole
Uchida, Leo
Buerano, Corazon C
Mwau, Matilu
Zha, Yan
Nie, Yingjie
Morita, Kouichi
author_facet Yu, Fuxun
Adungo, Ferdinard
Konongoi, Samson Limbaso
Inoue, Shingo
Sang, Rosemary
Ashur, Salame
Kwallah, Allan ole
Uchida, Leo
Buerano, Corazon C
Mwau, Matilu
Zha, Yan
Nie, Yingjie
Morita, Kouichi
author_sort Yu, Fuxun
collection PubMed
description BACKGROUND: Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis. To detect RVF virus (RVFV) infection, indirect immunoglobulin G (IgG) and immunoglobulin M (IgM) enzyme linked immunosorbent assays (ELISAs) which utilize recombinant RVFV nucleocapsid (RVFV-N) protein as assay antigen, have reportedly been used, however, there is still a need to develop more sensitive and specific methods of detection. METHODS: RVFV-N protein was expressed in Escherichia coli (E. coli) and purified by histidine-tag based affinity chromatography. This recombinant RVFV-N (rRVFV-N) protein was then used as antigen to develop an IgG sandwich ELISA and IgM capture ELISAs for human sera. Ninety six serum samples collected from healthy volunteers during the RVF surveillance programme in Kenya in 2013, and 93 serum samples collected from RVF-suspected patients during the 2006–2007 RVF outbreak in Kenya were used respectively, to evaluate the newly established rRVFV-N protein-based IgG sandwich ELISA and IgM capture ELISA systems in comparison with the inactivated virus-based ELISA systems. RESULTS: rRVFV-N protein-based-IgG sandwich ELISA and IgM capture ELISA for human sera were established. Both the new ELISA systems were in 100% concordance with the inactivated virus-based ELISA systems, with a sensitivity and specificity of 100%. CONCLUSIONS: Recombinant RVFV-N is a safe and affordable antigen for RVF diagnosis. Our rRVFV-N-based ELISA systems are safe and reliable tools for diagnosis of RVFV infection in humans and especially useful in large-scale epidemiological investigation and for application in developing countries.
format Online
Article
Text
id pubmed-6249750
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-62497502018-11-26 Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens Yu, Fuxun Adungo, Ferdinard Konongoi, Samson Limbaso Inoue, Shingo Sang, Rosemary Ashur, Salame Kwallah, Allan ole Uchida, Leo Buerano, Corazon C Mwau, Matilu Zha, Yan Nie, Yingjie Morita, Kouichi Virol J Research BACKGROUND: Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis. To detect RVF virus (RVFV) infection, indirect immunoglobulin G (IgG) and immunoglobulin M (IgM) enzyme linked immunosorbent assays (ELISAs) which utilize recombinant RVFV nucleocapsid (RVFV-N) protein as assay antigen, have reportedly been used, however, there is still a need to develop more sensitive and specific methods of detection. METHODS: RVFV-N protein was expressed in Escherichia coli (E. coli) and purified by histidine-tag based affinity chromatography. This recombinant RVFV-N (rRVFV-N) protein was then used as antigen to develop an IgG sandwich ELISA and IgM capture ELISAs for human sera. Ninety six serum samples collected from healthy volunteers during the RVF surveillance programme in Kenya in 2013, and 93 serum samples collected from RVF-suspected patients during the 2006–2007 RVF outbreak in Kenya were used respectively, to evaluate the newly established rRVFV-N protein-based IgG sandwich ELISA and IgM capture ELISA systems in comparison with the inactivated virus-based ELISA systems. RESULTS: rRVFV-N protein-based-IgG sandwich ELISA and IgM capture ELISA for human sera were established. Both the new ELISA systems were in 100% concordance with the inactivated virus-based ELISA systems, with a sensitivity and specificity of 100%. CONCLUSIONS: Recombinant RVFV-N is a safe and affordable antigen for RVF diagnosis. Our rRVFV-N-based ELISA systems are safe and reliable tools for diagnosis of RVFV infection in humans and especially useful in large-scale epidemiological investigation and for application in developing countries. BioMed Central 2018-11-22 /pmc/articles/PMC6249750/ /pubmed/30466469 http://dx.doi.org/10.1186/s12985-018-1071-y Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Yu, Fuxun
Adungo, Ferdinard
Konongoi, Samson Limbaso
Inoue, Shingo
Sang, Rosemary
Ashur, Salame
Kwallah, Allan ole
Uchida, Leo
Buerano, Corazon C
Mwau, Matilu
Zha, Yan
Nie, Yingjie
Morita, Kouichi
Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens
title Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens
title_full Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens
title_fullStr Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens
title_full_unstemmed Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens
title_short Comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens
title_sort comparison of enzyme-linked immunosorbent assay systems using rift valley fever virus nucleocapsid protein and inactivated virus as antigens
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6249750/
https://www.ncbi.nlm.nih.gov/pubmed/30466469
http://dx.doi.org/10.1186/s12985-018-1071-y
work_keys_str_mv AT yufuxun comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT adungoferdinard comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT konongoisamsonlimbaso comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT inoueshingo comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT sangrosemary comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT ashursalame comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT kwallahallanole comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT uchidaleo comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT bueranocorazonc comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT mwaumatilu comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT zhayan comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT nieyingjie comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens
AT moritakouichi comparisonofenzymelinkedimmunosorbentassaysystemsusingriftvalleyfevervirusnucleocapsidproteinandinactivatedvirusasantigens