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Journal of Clinical Microbiology, June 2000, p. 2324-2329, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Diagnostic Potential of Puumala Virus Nucleocapsid Protein Expressed in Drosophila melanogaster Cells

Katarina Brus Sjölander,1 Irina Golovljova,2 Alexander Plyusnin,3,4 and Åke Lundkvist1,3,*

Swedish Institute for Infectious Disease Control, S-171 82 Stockholm,1 and Microbiology and Tumor Biology Center, Karolinska Institute, S-171 77 Stockholm,3 Sweden; Institute of Preventive Medicine, Virus Ecology Laboratory, Tallinn EE0006, Estonia2; and Haartman Institute, Department of Virology, FIN-00014 University of Helsinki, Helsinki, Finland4

Received 2 December 1999/Returned for modification 7 February 2000/Accepted 25 March 2000

Puumala virus (PUU) nucleocapsid protein (N) was expressed in insect cells by using the Drosophila Expression System (DES; Invitrogen BV, Groningen, The Netherlands). Stable transfectants were established by hygromycin B selection and showed continuous expression of the recombinant protein (DES-PUU-N) for at least 5 months. The antigenic property of DES-PUU-N was shown to be identical to that of native PUU N when examined with a panel of hantavirus-specific monoclonal antibodies. Enzyme-linked immunosorbent assays (ELISAs) for detection of human immunoglobulin M (IgM) and IgG antibodies were established by using DES-PUU-N as antigen and were compared to assays based on native N. The ELISAs were evaluated for patient diagnosis and seroepidemiological purposes with panels of sera collected from patients with hemorrhagic fever with renal syndrome (HFRS) and from healthy blood donors. Equally high sensitivities and specificities for detection of PUU-specific IgM in acute-phase HFRS patient sera were obtained by the ELISA based on DES-PUU-N and the assay based on the native antigen. For detection of PUU-specific IgG, the ELISA based on monoclonal antibody-captured DES-PUU-N antigen showed optimal sensitivity and specificity.

* Corresponding author. Mailing address: Swedish Institute for Infectious Disease Control, Department of Virology, Tomtebodavägen 12B, S-171 82 Stockholm, Sweden. Phone: 46-8-4572641. Fax: 46-8-314744. E-mail: akelun{at}mbox.ki.se.

Journal of Clinical Microbiology, June 2000, p. 2324-2329, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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