Virus Genes

, 43:177 | Cite as

Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein

  • Marc Mertens
  • Eveline Kindler
  • Petra Emmerich
  • Jutta Esser
  • Christiane Wagner-Wiening
  • Roman Wölfel
  • Rasa Petraityte-Burneikiene
  • Jonas Schmidt-Chanasit
  • Aurelija Zvirbliene
  • Martin H. Groschup
  • Gerhard Dobler
  • Martin Pfeffer
  • Gerald Heckel
  • Rainer G. Ulrich
  • Sandra S. Essbauer


Puumala virus (PUUV) is the predominant hantavirus species in Germany causing large numbers of mild to moderate cases of haemorrhagic fever with renal syndrome (HFRS). During an outbreak in South-East Germany in 2004 a novel PUUV subtype designated Bavaria was identified as the causative agent of HFRS in humans [1]. Here we present a molecular characterization of this PUUV strain by investigating novel partial and almost entire nucleocapsid (N) protein-encoding small (S-) segment sequences and partial medium (M-) segment sequences from bank voles (Myodes glareolus) trapped in Lower Bavaria during 2004 and 2005. Phylogenetic analyses confirmed their classification as subtype Bavaria, which is further subdivided into four geographical clusters. The entire N protein, harbouring an amino-terminal hexahistidine tag, of the Bavarian strain was produced in yeast Saccharomyces cerevisiae and showed a slightly different reactivity with N-specific monoclonal antibodies, compared to the yeast-expressed N protein of the PUUV strain Vranica/Hällnäs. Endpoint titration of human sera from different parts of Germany and from Finland revealed only very slight differences in the diagnostic value of the different recombinant proteins. Based on the novel N antigen indirect and monoclonal antibody capture IgG-ELISAs were established. By using serum panels from Germany and Finland their validation demonstrated a high sensitivity and specificity. In summary, our investigations demonstrated the Bavarian PUUV strain to be genetically divergent from other PUUV strains and the potential of its N protein for diagnostic applications.


Hantavirus Puumala virus Phylogeny Recombinant nucleocapsid protein ELISA Diagnostics 



We would like to thank Ausra Razanskiene and Kestutis Sasnauskas (Vilnius) and Bernd Köllner (Riems) for continous support and helpful discussions, Åke Lundkvist (Stockholm), Kumiko Yoshimatsu and Jiro Arikawa (Sapporo) for provision of N protein-specific mAbs and Olli Vapalahti (Helsinki), Regina Allwinn and Holger F. Rabenau (Frankfurt) for provision of human serum panels. We kindly acknowledge Harald Weber, Aileene Lorber, Rahime Terzioglu, Elmar Schröpfer, Ralf Hagen, Mirko Köhler, Anne Grumbach, Peter Klein and Kerstin Weiss for their support during rodent trapping, rodent necropsy and collection of human sera in South-East Germany. The excellent technical lab assistance of Jana Blumhardt, Ina Römer, Dörte Kaufmann, Sabine Fink and Astrid Thomas is gratefully acknowledged. The work was supported by Research-Project M/SAB1/5/A017 for the Bundeswehr Medical Service to RGU.

Supplementary material

11262_2011_620_MOESM1_ESM.pptx (64 kb)
Supplementary Figure. Bayesian reconstruction of phylogenetic relationships based on 968 bp mtDNA (cyt b) from 120 Myodes glareolus with M. rufocanus as outgroup. Only support values for main nodes that connect major evolutionary lineages are displayed. Posterior probabilities are indicated above the major branches and percentage of bootstrap support for neighbour-joining (NJ) algorithms below the branches. * indicates a different topology based on NJ algorithms, ns refers to posterior probabilities <0.50 and percentages of bootstrap support <50%. Arrows depict cyt b sequences from bank voles from Bavaria where PUUV sequences were detected in (PPTX 64 kb)


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Marc Mertens
    • 1
  • Eveline Kindler
    • 2
  • Petra Emmerich
    • 3
  • Jutta Esser
    • 4
  • Christiane Wagner-Wiening
    • 5
  • Roman Wölfel
    • 6
  • Rasa Petraityte-Burneikiene
    • 7
  • Jonas Schmidt-Chanasit
    • 3
  • Aurelija Zvirbliene
    • 7
  • Martin H. Groschup
    • 1
  • Gerhard Dobler
    • 8
  • Martin Pfeffer
    • 9
  • Gerald Heckel
    • 2
    • 10
  • Rainer G. Ulrich
    • 1
  • Sandra S. Essbauer
    • 8
  1. 1.Institute for Novel and Emerging Infectious DiseasesFriedrich-Loeffler-Institut, Federal Research Institute for Animal HealthGreifswald-Insel RiemsGermany
  2. 2.Computational and Molecular Population Genetics (CMPG), Institute of Ecology and EvolutionUniversity of BernBernSwitzerland
  3. 3.Diagnostic Virology LaboratoryBernhard-Nocht-Institute for Tropical MedicineHamburgGermany
  4. 4.Labor Enzenauer und Kollegen, Abteilung SerologieOsnabrückGermany
  5. 5.Landesgesundheitsamt Baden-Württemberg, Regierungspräsidium StuttgartStuttgartGermany
  6. 6.Department of Medical Bioreconnaissance and VerificationBundeswehr Institute of MicrobiologyMunichGermany
  7. 7.VU Institute of BiotechnologyVilniusLithuania
  8. 8.Department of Virology & RickettsiologyBundeswehr Institute of MicrobiologyMunichGermany
  9. 9.Institute of Animal Hygiene and Veterinary Public Health, Veterinary FacultyUniversity of LeipzigLeipzigGermany
  10. 10.Swiss Institute of Bioinformatics (SIB)LausanneSwitzerland

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