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Clinical and Vaccine Immunology, December 2006, p. 1349-1357, Vol. 13, No. 12
1071-412X/06/$08.00+0     doi:10.1128/CVI.00208-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Development of Novel Immunoglobulin G (IgG), IgA, and IgM Enzyme Immunoassays Based on Recombinant Puumala and Dobrava Hantavirus Nucleocapsid Proteins{triangledown}

Helga Meisel,1* Anne Wolbert,1,{dagger} Ausra Razanskiene,2 Andreas Marg,1,{ddagger} Andris Kazaks,3 Kestutis Sasnauskas,2 Georg Pauli,4 Rainer Ulrich,1,§ and Detlev H. Krüger1

Institute of Virology, Helmut-Ruska-Haus, Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany,1 Institute of Biotechnology, Graiciuno 8, 02241 Vilnius, Lithuania,2 Biomedical Research and Study Centre, University of Latvia, Ratsupites Str. 1, 1067 Riga, Latvia,3 Robert-Koch-Institut, Nordufer 20, 13353 Berlin, Germany4

Received 31 May 2006/ Returned for modification 9 July 2006/ Accepted 28 September 2006

Human infections with Asian and European hantaviruses can result in hemorrhagic fever with renal syndromes of differing severities characterized by renal dysfunction and sometimes by pulmonary symptoms. For the serological detection of human infections by hantaviruses relevant for Europe, we developed monoclonal antibody capture immunoglobulin G (IgG) and IgA enzyme-linked immunosorbent assays (ELISAs) based on yeast-expressed nucleocapsid proteins of Puumala and Dobrava hantaviruses. Moreover, for diagnosis of acute infections, µ-capture IgM ELISAs were established with nucleocapsid proteins expressed in Drosophila melanogaster Schneider S2 cells. The cutoff values of the ELISAs were determined by investigation of up to 500 human anti-hantavirus-negative serum samples. The specificities of the Puumala and Dobrava virus-specific IgM, IgA, and IgG ELISAs were found to be 100%. The sensitivities of these ELISAs were determined to be 100% with panels of characterized anti-Puumala or anti-Dobrava virus-positive human serum samples. In most cases, Puumala and Dobrava virus infections could be differentiated by ELISA reactivity alone, i.e., endpoint titration with homologous and heterologous antigens.

* Corresponding author. Mailing address: Institute of Virology, Helmut-Ruska-Haus, Charité-Universitätsmedizin Berlin, Campus Charité Mitte, Charitéplatz 1, 10117 Berlin, Germany. Phone: 49-30-450-525141. Fax: 49-30-450-525907. E-mail: helga.meisel{at}charite.de.

{triangledown} Published ahead of print on 4 October 2006.

{dagger} Present address: Robert-Koch-Institut, Nordufer 20, 13353 Berlin, Germany.

{ddagger} Present address: Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Str. 10, 13125 Berlin, Germany.

§ Present address: Friedrich-Loeffler-Institut, Institute for Novel and Emerging Infectious Diseases, Boddenblick 5 a, 17493 Greifswald-Insel Riems, Germany.

Clinical and Vaccine Immunology, December 2006, p. 1349-1357, Vol. 13, No. 12
1071-412X/06/$08.00+0     doi:10.1128/CVI.00208-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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