Enzyme-linked immunosorbent assays for detection of antibodies to Ebola and Marburg viruses using recombinant nucleoproteins

M. Saijo, M. Niikura, S. Morikawa, Thomas Ksiazek, R. F. Meyer, C. J. Peters, I. Kurane

Research output: Contribution to journalArticle

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Abstract

The full-length nucleoprotein (NP) of Ebola virus (EBO) was expressed as a His-tagged recombinant protein (His-EBO-NP) by a baculovirus system. Carboxy-terminal halves of NPs of EBO and Marburg virus (MBG) were expressed as glutathione S-transferase-tagged recombinant proteins in an Escherichia coli system. The antigenic regions on the NPs of EBO and MBG were determined by both Western blotting and enzyme-linked immunosorbent assay (ELISA) to be located on file C-terminal halves. The C-terminal 110 and 102 amino acids of the NPs of EBO and MBG, respectively, possess strong antigenicity. The full-length NP of EBO was strongly expressed in insect cells upon infection with the recombinant baculovirus, while expression of the full-length NP of MBG was weak. We developed an immunoglobulin G (IgG) ELISA using His-EBO-NP and the C-terminal halves of the NPs of EBO and MBG as antigens. We evaluated the IgG ELISA for the ability to detect IgG antibodies to EBO and MBG, using human sera collected from EBO and MBG patients. The IgG ELISA with the recombinant NPs showed high sensitivity and specificity in detecting EBO and MBG antibodies. The results indicate that ELISA systems prepared with the recombinant NPs of EBO and MBG are valuable tools for the diagnosis of EBO and MBG infections and for seroepidemiological field studies.

Original languageEnglish (US)
Pages (from-to)1-7
Number of pages7
JournalJournal of Clinical Microbiology
Volume39
Issue number1
DOIs
StatePublished - 2001
Externally publishedYes

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Marburgvirus
Ebolavirus
Nucleoproteins
Enzyme-Linked Immunosorbent Assay
Antibodies
Immunoglobulin G
Baculoviridae
Recombinant Proteins
Seroepidemiologic Studies
Virus Diseases

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Enzyme-linked immunosorbent assays for detection of antibodies to Ebola and Marburg viruses using recombinant nucleoproteins. / Saijo, M.; Niikura, M.; Morikawa, S.; Ksiazek, Thomas; Meyer, R. F.; Peters, C. J.; Kurane, I.

In: Journal of Clinical Microbiology, Vol. 39, No. 1, 2001, p. 1-7.

Research output: Contribution to journalArticle

Saijo, M. ; Niikura, M. ; Morikawa, S. ; Ksiazek, Thomas ; Meyer, R. F. ; Peters, C. J. ; Kurane, I. / Enzyme-linked immunosorbent assays for detection of antibodies to Ebola and Marburg viruses using recombinant nucleoproteins. In: Journal of Clinical Microbiology. 2001 ; Vol. 39, No. 1. pp. 1-7.
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