title: Recombinant Ebola virus nucleoprotein and glycoprotein (Gabon 94 strain) provide new tools for the detection of human infections.
abstract: After cloning and sequencing the glycoprotein (GP) gene of one of the Gabonese strains of Ebola virus isolated during the 1994-1996 outbreak, it was shown that the circulating virus was of the Zaire subtype. This was confirmed in this study by cloning and sequencing the nucleoprotein (NP) gene of this strain. These two structural proteins were also expressed as recombinant proteins and used in ELISA tests. NP was expressed as a His-tagged fusion protein in Escherichia coli and was purified on resins charged with nickel ions. GP was expressed by means of recombinant baculoviruses in Spodoptera frugiperda cells. Both recombinant proteins reacted positively in ELISAs for the detection of IgG antibodies in convalescent human sera from Gabon and Zaire. The difference in the relative titres of anti-NP and -GP antibodies was variable, depending on the sera. In addition, the recombinant NP reacted with heterologous sera from Côte d'Ivoire and was used successfully to detect IgM antibodies by mu-capture ELISA in sera from Gabonese patients.
authors.affiliation: Unité des Arbovirus et virus des Fièvres Hémorragiques, Institut Pasteur, Paris, France.
authors.forename: C
authors.initials: C
authors.lastname: Prehaud
authors.forename: E
authors.initials: E
authors.lastname: Hellebrand
authors.forename: D
authors.initials: D
authors.lastname: Coudrier
authors.forename: V E
authors.initials: VE
authors.lastname: Volchkov
authors.forename: V A
authors.initials: VA
authors.lastname: Volchkova
authors.forename: H
authors.initials: H
authors.lastname: Feldmann
authors.forename: B
authors.initials: B
authors.lastname: Le Guenno
authors.forename: M
authors.initials: M
authors.lastname: Bouloy
mesh_terms: D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D029043:Ebolavirus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D005814:Genes, Viral; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008969:Molecular Sequence Data; D009698:Nucleoproteins / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D007074:Immunoglobulin G; D009698:Nucleoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins