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Detection and sequencing of new french Puumala virus isolates (PALADIN project)
Guillaume Castel, S. Murri, F. Chevenet, Sarah Madrières, J. Vulin, Caroline Tatard, Maxime Galan, Laure Benoit, Anne Loiseau, Philippe Marianneau, et
al.
To cite this version:
Guillaume Castel, S. Murri, F. Chevenet, Sarah Madrières, J. Vulin, et al.. Detection and sequencing of new french Puumala virus isolates (PALADIN project). 11. International Conference on Hantaviruses, Sep 2019, Leuven, Belgium. 2019. �hal-02790435�
Detec%on and sequencing of new french Puumala virus isolates (PALADIN project)
Guillaume Castel
1, Séverine Murri
2, François Chevenet
3, Sarah Madrières
1,2, Johann Vulin
2, Caroline Tatard
1, Maxime Galan
1, Laure Benoit
1, Anne Loiseau
1, Philippe Marianneau
2, Nathalie Charbonnel
11CBGP, INRA, CIRAD, IRD, Montpellier SupAgro, Univ Montpellier, Montpellier, France; 2Laboratoire de Lyon, ANSES, Unité de virologie, Lyon, France; 3MIVEGEC & LIRMM, Université de Montpellier, CNRS, IRD, France.
Context Methods
• Puumala virus (PUUV) is the causa%ve agent of nephropathia epidemica (NE). The PALADIN project aims to understand the evolu%on and current diversity of PUUV circula%ng in France in bank vole popula%ons.
• Samples from Franche-‐Comté, Jura, Aube, Alsace, Ardennes (endemic regions for NE) and Loiret (non endemic region) are regularly harvested since 2009. Dis%nct PUUV
"sublineages" circulate in these regions.
• We sample bank voles in in-‐between forests where PUUV has not been previously inves%gated whereas recent human cases have been reported in their vicinity (Reynes et al, 2019).
• We inves%gate some of the gene%c features accompanying / favoring PUUV pathogenicity in humans, and their poten%al geographic varia%ons.
• We present here the first results of this project concerning the forest of Morvan.
• Analyses :
1) Detec%on: Serology & qRT-‐PCR (lung)
2) Nested RT-‐PCR (N gene) + Sanger sequencing 3) Phylogene%c studies
Ø PUUV circulates within bank vole popula%ons in the forest of Morvan (considered as a non endemic region).
Ø Isolates circula%ng in bank voles from Morvan correspond to the strain isolated from a recent human NE case and are phylogene%cally closed from the isolates of the non endemic Loiret region.
Ø Isolates from Morvan share one of the AA signature specific of isolates from Loiret otherwise they are similar to those of Jura/Vosges/
Alsace/Aube. The role of these specific AA in the pathogenicity of the different isolates needs more inves%ga%ons.
Conclusions
Results - Forest of Morvan
Email: [email protected]
Figure.1: phylogeny (Maximum Likelihood) of french PUUV isolates (S segment).
Branch supports > 0.8 are indicated by a red point.
2) PUUV isolates from bank voles of Morvan and from the recent first human case in this region (★) are phylogene%caly closed to those from the non endemic region of Loiret (Fig. 1).
1) PUUV detec%on: 23% of the sampled bank voles were posi%ve (serology & qRT-‐PCR) for PUUV (3/13).
3) Isolates from Morvan share one of the three AA signatures specific of Loiret (non endemic) and none of those specific of Jura/Vosges/
Alsace/Aube and of Ardennes (endemic).
Table.1: Specific Amino-‐acids signatures of PUUV french
isolates on the N protein.
Acknowledgments (sampling):
-‐ Sylvain Piry
-‐ Nicolas Leménager -‐ Emmanuelle ArBge -‐ Philippe Audiot -‐ Julien Pradel
• Rodent trapping with blood and organs sampling
64 79 233 258 294 305
Ardennes Q T A Q S A
Jura/Vosges/Alsaces/Aube R T T Q S S
Morvan R T A K S S
Loiret R A A K A S
Isolates
AminoFacidsGsignaturesGonGNGprotein
