Medical Microbiology and Immunology

, Volume 198, Issue 2, pp 83–91 | Cite as

Seroepidemiological study in a Puumala virus outbreak area in South-East Germany

  • Marc Mertens
  • Roman Wölfel
  • Katrin Ullrich
  • Kumiko Yoshimatsu
  • Jana Blumhardt
  • Ina Römer
  • Jutta Esser
  • Jonas Schmidt-Chanasit
  • Martin H. Groschup
  • Gerhard Dobler
  • Sandra S. Essbauer
  • Rainer G. Ulrich
Original Investigation


Puumala virus (PUUV) is the cause of the majority of haemorrhagic fever with renal syndrome cases in Germany. In 2004, a nephropathia epidemica outbreak was recorded in Lower Bavaria, South-East Germany. For a seroepidemiological study in this region including the resident population at four locations (n = 178) and soldiers from one location (n = 208) indirect immunoglobulin M (IgM) and immunoglobulin G (IgG) enzyme-linked immunosorbent assays (ELISAs) and immunoblot tests based on a yeast-expressed PUUV nucleocapsid protein were established. The validation using human serum panels originating from Germany revealed a diagnostic sensitivity and specificity of 98/100% for the IgM ELISA, 99/99% for the IgG ELISA, 99/100% for the IgM immunoblot test and 100/96% for the IgG immunoblot test. Using the novel IgG assays as well as a commercial IgG ELISA and an immunofluorescence assay for the resident population an average prevalence of 6.7% (12 of 178) with a range of 0% (0 of 21) to 11.9% (7 of 59) was observed. Positive serological results were equally distributed between males and females with an average age of 63 for males and 52 for females. The seroprevalence in the soldier group was found to be about 1% with one positive male of 203 (age 46 years) and one positive female of five (age 47 years). In conclusion, the PUUV seroprevalence in the residents of the outbreak region in Lower Bavaria was found to be up to fivefold higher than the average hantavirus seroprevalence of the German population.


Hantavirus Puumala virus Germany Nucleocapsid protein ELISA Immunoblot Seroprevalence 



We thank Rasa Petraityte, Rita Vorobjoviene, Ausra Razanskiene, Kestutis Sasnauskas (Vilnius), Claudia Dettmer (Wusterhausen) for support and Bernd Köllner for helpful discussions. Human sera for test establishment and validation were kindly provided by Hans Quast (Schledehausen), Regina Allwinn, Holger F. Rabenau (Frankfurt), Silvia Dorn (Neuried) and Petra Emmerich (Hamburg). TULV, strain Moravia, and PUUV, strain CG18-20, were obtained from Alexander Plyusnin (Helsinki) and the WHO collaboration centre (Seoul). We also thank the soldiers of the Mobile Medical Biological Reconnaissance and Verification Unit from the Bundeswehr Institute of Microbiology (Munich) as well as Rainer Stern (Geiersthal), Wolfgang Blank (Kirchberg), Irmengard Schoder (Schöfweg) and Karl-Heinz Haberer (Spiegelau) who conducted the serosurvey. The excellent technical assistance of Peter Klein, Anne Grumbach and Astrid Thomas is acknowledged. The work was supported by Research-Project M/SAB1/5/A017 for the Bundeswehr Medical Service. Jonas Schmidt-Chanasit acknowledges support by Förderverein of the Friedrich-Loeffler-Institut and Paul und Ursula Klein-Stiftung.


  1. 1.
    Aberle SW, Ecker M, Aberle JH, Arneitz K, Khanakah G, Radda A, Radda I, Popow-Kraupp T, Kunz C, Heinz FX (1999) Nephropathia epidemica and Puumala virus in Austria. Eur J Clin Microbiol Infect Dis 18:467–472. doi:10.1007/s100960050325 PubMedCrossRefGoogle Scholar
  2. 2.
    Antoniadis A, Pilaski J, Klein H, Zöller L, Gorschewski O (1985) Serologic evidence for Korean Haemorrhagic Fever in Western Germany. Zbl Bakt Hyg A 260:459Google Scholar
  3. 3.
    Araki K, Yoshimatsu K, Ogino M, Ebihara H, Lundkvist A, Kariwa H, Takashima I, Arikawa J (2001) Truncated hantavirus nucleocapsid proteins for serotyping Hantaan, Seoul, and Dobrava hantavirus infections. J Clin Microbiol 39:2397–2404. doi:10.1128/JCM.39.7.2397-2404.2001 PubMedCrossRefGoogle Scholar
  4. 4.
    Clement J, Underwood P, Ward D, Pilaski J, LeDuc J (1996) Hantavirus outbreak during military manoeuvres in Germany. Lancet 347:336. doi:10.1016/S0140-6736(96)90519-X PubMedCrossRefGoogle Scholar
  5. 5.
    Dargeviciute A, Brus SK, Sasnauskas K, Krüger DH, Meisel H, Ulrich R, Lundkvist Å (2002) Yeast-expressed Puumala hantavirus nucleocapsid protein induces protection in a bank vole model. Vaccine 20:3523–3531. doi:10.1016/S0264-410X(02)00341-9 PubMedCrossRefGoogle Scholar
  6. 6.
    Essbauer S, Schmidt J, Conraths FJ, Friedrich R, Koch J, Hautmann W, Pfeffer M, Wölfel R, Finke J, Dobler G, Ulrich R (2006) A new Puumala hantavirus subtype in rodents associated with an outbreak of Nephropathia epidemica in South-East Germany in 2004. Epidemiol Infect 134:1333–1344. doi:10.1017/S0950268806006170 PubMedCrossRefGoogle Scholar
  7. 7.
    Essbauer S, Schmidt-Chanasit J, Madeja EL, Wegener W, Friedrich R, Petraityte R, Sasnauskas K, Jacob J, Koch J, Dobler G, Conraths FJ, Pfeffer M, Pitra C, Ulrich RG (2007) Nephropathia epidemica outbreak in a metropolitan area, Germany. Emerg Infect Dis 13:1271–1273PubMedGoogle Scholar
  8. 8.
    Feldmann H, Sanchez A, Morzunov S, Spiropoulou CF, Rollin PE, Ksiazek TG, Peters CJ, Nichol ST (1993) Utilization of autopsy RNA for the synthesis of the nucleocapsid antigen of a newly recognized virus associated with hantavirus pulmonary syndrome. Virus Res 30:351–367. doi:10.1016/0168-1702(93)90101-R PubMedCrossRefGoogle Scholar
  9. 9.
    Golovljova I, Sjölander KB, Lindegren G, Vene S, Vasilenko V, Plyusnin A, Lundkvist A (2002) Hantaviruses in Estonia. J Med Virol 68:589–598. doi:10.1002/jmv.10231 PubMedCrossRefGoogle Scholar
  10. 10.
    Heyman P, Vervoort T, Colson P, Chu YK, Avsic-Zupanc T, Lundkvist A (1999) A major outbreak of hantavirus infection in Belgium in 1995 and 1996. Epidemiol Infect 122:447–453. doi:10.1017/S0950268899002344 PubMedCrossRefGoogle Scholar
  11. 11.
    Hofmann J, Meisel H, Klempa B, Vesenbeckh SM, Beck R, Michel D, Schmidt-Chanasit J, Ulrich RG, Grund S, Enders G, Krüger DH (2008) Molecular epidemiology of a large hantavirus outbreak in Germany, 2007. Emerg Infect Dis 14:850–852PubMedCrossRefGoogle Scholar
  12. 12.
    Kimmig P, Silva-González R, Backe H, Brockmann S, Oehme R, Ernst E, Mackenstedt U (2001) Epidemiology of hantaviruses in Baden-Wurttemberg. Gesundheitswesen 63:107–112. doi:10.1055/s-2001-10961 PubMedCrossRefGoogle Scholar
  13. 13.
    Klempa B, Meisel H, Räth S, Bartel J, Ulrich R, Krüger DH (2003) Occurrence of renal and pulmonary syndrome in a region of North-East Germany where Tula hantavirus circulates. J Clin Microbiol 41:4894–4897. doi:10.1128/JCM.41.10.4894-4897.2003 PubMedCrossRefGoogle Scholar
  14. 14.
    Klempa B, Schütt M, Auste B, Ulrich R, Meisel H, Krüger DH (2004) First molecular identification of human Dobrava virus infection in Central Europe. J Clin Microbiol 42:1322–1325. doi:10.1128/JCM.42.3.1322-1325.2004 PubMedCrossRefGoogle Scholar
  15. 15.
    Klingström J, Lindgren T, Ahlm C (2008) Sex-dependent differences in plasma cytokine responses to hantavirus infection. Clin Vaccine Immunol 15:885–887. doi:10.1128/CVI.00035-08 PubMedCrossRefGoogle Scholar
  16. 16.
    Lledó L, Gegúndez MI, Saz JV, Alves MJ, Beltrán M (2002) Serological study of hantavirus in man in the autonomous community of Madrid, Spain. J Med Microbiol 51:861–865PubMedGoogle Scholar
  17. 17.
    Mailles A, Abu Sin M, Ducoffre G, Heyman P, Koch J, Zeller H (2005) Larger than usual increase in cases of hantavirus infections in Belgium, France and Germany, June 2005. Eurosurveillance Wkly 10:10–18Google Scholar
  18. 18.
    Meisel H, Wolbert A, Razanskiene A, Marg A, Kazaks A, Sasnauskas K, Pauli G, Ulrich R, Krüger DH (2006) Development of novel immunoglobulin G (IgG), IgA, and IgM enzyme immunoassays based on recombinant Puumala and Dobrava hantavirus nucleocapsid proteins. Clin Vaccine Immunol 13:1349–1357. doi:10.1128/CVI.00208-06 PubMedCrossRefGoogle Scholar
  19. 19.
    Mustonen J, Vapalahti O, Henttonen H, Pasternack A, Vaheri A (1998) Epidemiology of hantavirus infections in Europe. Nephrol Dial Transpl 13:2729–2731. doi:10.1093/ndt/13.11.2729 CrossRefGoogle Scholar
  20. 20.
    Niklasson B, Leduc J, Nyström K, Nyman L (1987) Nephropathia epidemica: incidence of clinical cases and antibody prevalence in an endemic area of Sweden. Epidemiol Infect 99:559–562PubMedCrossRefGoogle Scholar
  21. 21.
    Padula PJ, Rossi CM, Della Valle MO, Martínez PV, Colavecchia SB, Edelstein A, Miguel SD, Rabinovich RD, Segura EL (2000) Development and evaluation of a solid-phase enzyme immunoassay based on Andes hantavirus recombinant nucleoprotein. J Med Microbiol 49:149–155PubMedGoogle Scholar
  22. 22.
    Papa A, Antoniadis A (2001) Hantavirus infections in Greece—an update. Eur J Epidemiol 17:189–194. doi:10.1023/A:1017987104363 PubMedCrossRefGoogle Scholar
  23. 23.
    Petraityte R, Jin L, Hunjan R, Razanskiene A, Zvirbliene A, Sasnauskas K (2007) Use of Saccharomyces cerevisiae-expressed recombinant nucleocapsid protein to detect Hantaan virus-specific immunoglobulin G (IgG) and IgM in oral fluid. Clin Vaccine Immunol 14:1603–1608. doi:10.1128/CVI.00188-07 PubMedCrossRefGoogle Scholar
  24. 24.
    Petraityte R, Yang H, Hunjan R, Razanskiene A, Dhanilall P, Ulrich RG, Sasnauskas K, Jin L (2008) Development and evaluation of serological assays for detection of Hantaan virus-specific antibodies in human sera using yeast-expressed nucleocapsid protein. J Virol Methods 148:89–95. doi:10.1016/j.jviromet.2007.10.019 PubMedCrossRefGoogle Scholar
  25. 25.
    Pilaski J, Ellerich C, Kreutzer T, Benik W, Lewandowski B, Lang A, Autenrieth IB, Vanek E (1991) Endemisches Vorkommen des Hämorrhagischen Fiebers mit renalem Syndrom (HFRS) in der Bundesrepublik Deutschland. Z Arztl Fortbild (Jena) 85:869–874Google Scholar
  26. 26.
    Rose AM, Vapalahti O, Lyytikäinen O, Nuorti P (2003) Patterns of Puumala virus infection in Finland. Euro Surveill 8:9–13PubMedGoogle Scholar
  27. 27.
    Sandmann S, Meisel H, Razanskiene A, Wolbert A, Pohl B, Krüger DH, Sasnauskas K, Ulrich R (2005) Detection of human hantavirus infections in Lithuania. Infection 33:66–72. doi:10.1007/s15010-005-4058-8 PubMedCrossRefGoogle Scholar
  28. 28.
    Schilling S, Emmerich P, Klempa B, Auste B, Schnaith E, Schmitz H, Krüger DH, Günther S, Meisel H (2007) Hantavirus outbreak in Germany: limitations of routine serological diagnostics and clustering of virus sequences of human and rodent origin. J Clin Microbiol 45:3008–3014. doi:10.1128/JCM.02573-06 PubMedCrossRefGoogle Scholar
  29. 29.
    Schmidt J, Meisel H, Hjelle B, Krüger DH, Ulrich R (2005) Development and evaluation of serological assays for detection of human hantavirus infections caused by Sin Nombre virus. J Clin Virol 33:247–253PubMedGoogle Scholar
  30. 30.
    Schmidt J, Meisel H, Capria SG, Petraityte R, Lundkvist Å, Hjelle B, Vial PA, Padula P, Krüger DH, Ulrich R (2006) Serological assays for the detection of human Andes hantavirus infections based on its yeast-expressed nucleocapsid protein. Intervirology 49:173–184. doi:10.1159/000089378 PubMedCrossRefGoogle Scholar
  31. 31.
    Sibold C, Ulrich R, Labuda M, Lundkvist Å, Martens H, Schütt M, Gerke P, Leitmeyer K, Meisel H, Krüger DH (2001) Dobrava hantavirus causes hemorrhagic fever with renal syndrome in central Europe and is carried by two different Apodemus mice species. J Med Virol 63:158–167. doi:10.1002/1096-9071(20000201)63:2<158::AID-JMV1011>3.0.CO;2-#PubMedCrossRefGoogle Scholar
  32. 32.
    Ulrich R, Meisel H, Schütt M, Schmidt J, Kunz A, Klempa B, Niedrig M, Kimmig P, Pauli G, Krüger DH, Koch J (2004) Verbreitung von Hantavirusinfektionen in Deutschland. Bundesgesundheitsblatt 47:661–670. doi:10.1007/s00103-004-0858-8 CrossRefGoogle Scholar
  33. 33.
    Vapalahti O, Mustonen J, Lundkvist A, Henttonen H, Plyusnin A, Vaheri A (2003) Hantavirus infections in Europe. Lancet Infect Dis 3:653–661. doi:10.1016/S1473-3099(03)00774-6 PubMedCrossRefGoogle Scholar
  34. 34.
    Zöller L, Yang S, Gött P, Bautz EKF, Darai G (1993) Use of recombinant nucleocapsid proteins of the Hantaan and Nephropathia epidemica serotypes of hantaviruses as immunodiagnostic antigens. J Med Virol 39:200–207. doi:10.1002/jmv.1890390305 PubMedCrossRefGoogle Scholar
  35. 35.
    Zöller L, Faulde M, Meisel H, Ruh B, Kimmig P, Schelling U, Zeier M, Kulzer P, Becker C, Roggendorf M, Bautz EKF, Krüger DH, Darai G (1995) Seroprevalence of hantavirus antibodies in Germany as determined by a new recombinant enzyme immunoassay. Eur J Clin Microbiol Infect Dis 14:305–313. doi:10.1007/BF02116523 PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Marc Mertens
    • 1
  • Roman Wölfel
    • 2
  • Katrin Ullrich
    • 3
  • Kumiko Yoshimatsu
    • 4
  • Jana Blumhardt
    • 1
  • Ina Römer
    • 1
  • Jutta Esser
    • 5
  • Jonas Schmidt-Chanasit
    • 3
    • 6
    • 7
  • Martin H. Groschup
    • 1
  • Gerhard Dobler
    • 2
  • Sandra S. Essbauer
    • 2
  • Rainer G. Ulrich
    • 1
  1. 1.Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, OIE Collaborating Centre for Zoonoses in Europe, Institute for Novel and Emerging Infectious DiseasesGreifswald, Insel RiemsGermany
  2. 2.Bundeswehr Institute of MicrobiologyMunichGermany
  3. 3.Institute of EpidemiologyWusterhausenGermany
  4. 4.Institute for Animal Experimentation, Hokkaido University Graduate School of MedicineSapporoJapan
  5. 5.Labor Enzenauer und Kollegen, Abteilung SerologieOsnabrückGermany
  6. 6.Institute of Medical Virology, Hospital of the Johann Wolfgang Goethe UniversityFrankfurt am MainGermany
  7. 7.Department of VirologyBernhard-Nocht-Institute for Tropical MedicineHamburgGermany

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