Expression of the nucleoprotein of the Puumala virus from the recombinant Semliki Forest virus replicon: characterization and use as a potential diagnostic tool.
Puumala virus (Bunyaviridae family, Hantavirus genus) causes a mild form of hemorrhagic fever with renal syndrome (HFRS) called nephropathia epidemica in northern and central Europe. Serological tests are used for diagnosis, but antigen production is difficult because the virus grows poorly in tissue culture. We expressed the N protein (nucleoprotein) of Puumala virus via the Semliki Forest virus (SFV) replicon in mammalian cells and compared its antigenic properties with those of the native antigen derived from Puumala virus-infected cells. Detection of immunoglobulin G or immunoglobulin M by enzyme-linked immunosorbent assay (ELISA), micro -capture ELISA, and indirect immunofluorescence assay was (at least) as effective with the recombinant antigen as with the native antigen when HFRS patient sera or organ washes from wild rodents were tested. No nonspecific reaction was observed. Thus, the SFV-expressed N protein of Puumala virus appears as a valid antigen, specific and sensitive for serological investigations.
Billecocq
A
A
Unité de Génétique Moléculaire des Bunyaviridés, Institut Pasteur, 75724 Paris Cedex 15, France.
Coudrier
D
D
Boué
F
F
Combes
B
B
Zeller
H
H
Artois
M
M
Bouloy
M
M
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D003411:Arvicolinae / Q000821:virology; D006224:Cricetinae; D004197:Disease Reservoirs; D004797:Enzyme-Linked Immunosorbent Assay; D006480:Hemorrhagic Fever with Renal Syndrome / Q000097:blood / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis* / Q000276:immunology; D007075:Immunoglobulin M / Q000032:analysis* / Q000276:immunology; D008856:Microscopy, Fluorescence; D019251:Nucleocapsid / Q000096:biosynthesis* / Q000235:genetics; D019590:Nucleocapsid Proteins; D029262:Puumala virus / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012093:Replicon; D012672:Semliki forest virus / Q000235:genetics*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; C100886:nucleocapsid protein, Puumala virus