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Clinical and Diagnostic Laboratory Immunology, July 2003, p. 658-663, Vol. 10, No. 4
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.4.658-663.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Expression of the Nucleoprotein of the Puumala Virus from the Recombinant Semliki Forest Virus Replicon: Characterization and Use as a Potential Diagnostic Tool

A. Billecocq,1 D. Coudrier,1 F. Boué,2 B. Combes,3 H. Zeller,4 M. Artois,5 and M. Bouloy1*

Unité de Génétique Moléculaire des Bunyaviridés, Institut Pasteur, 75724 Paris Cedex 15,1 AFSSA Nancy,2 Entente Interdépartementale de Lutte contre la Rage et Autres Zoonoses, 54220 Malzéville Cedex,3 Centre National de Référence des Arbovirus et des Fièvres Hémorragiques Virales, Institut Pasteur, 69365 Lyon Cedex 7,4 Unité Pathologie Infectieuse, Ecole Nationale Vétérinaire de Lyon, 69280 Marcy l'Etoile, France5

Received 23 January 2003/ Returned for modification 17 April 2003/ Accepted 5 May 2003

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), µ-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.

* Corresponding author. Mailing address: Unité Postulante de Génétique Moléculaire des Bunyaviridés, Institut Pasteur, 25-28, rue du Docteur Roux, 75724 Paris Cedex 15, France. Phone: 33 0 1 40 61 31 57. Fax: 33 0 1 40 61 31 51. E-mail: mbouloy{at}pasteur.fr.

Clinical and Diagnostic Laboratory Immunology, July 2003, p. 658-663, Vol. 10, No. 4
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.4.658-663.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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