Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: First isolation and genetic characterization of Puumala orthohantavirus strains from France Puumala orthohantavirus (PUUV) causes a mild form of haemorrhagic fever with renal syndrome (HFRS) called nephropathia epidemica (NE), regularly diagnosed in Europe. France represents the western frontier of the expansion of NE in Europe with two distinct areas: an endemic area (north-eastern France) where PUUV circulates in rodent populations, with detection of many human NE cases, and a non-endemic area (south-western France) where the virus is not detected, with only a few human cases being reported. France is thus a relevant country in which to study the factors that influence the evolution of PUUV distribution. In this study, we describe for the first time the isolation of two PUUV strains from two distinct French geographical areas: Ardennes (endemic area) and Loiret (non-endemic area). To isolate PUUV efficiently, we selected wild bank voles (Myodes glareolus, the specific reservoir of PUUV) captured in these areas and that were seronegative for anti-PUUV IgG (ELISA), but showed a non-negligible viral RNA load in their lung tissue (qRT-PCR). With this study design, we were able to cultivate and maintain these two strains in Vero E6 cells and also propagate both strains in immunologically neutral bank voles efficiently and rapidly. Complete coding sequences of the S and M segments were determined by Sanger sequencing from RNA extracted from positive bank voles (naturally and experimentally infected) and from supernatants of Vero E6 cell extracts. For the M segment, nucleotide sequences were 100% identical for both strains. For the S segment, the amino-acid sequences from each strain revealed one mismatch between sequences obtained from tissue and from cell supernatants, revealing distinct "bank vole" and a "cell" molecular profile. High-throughput sequencing confirmed Sanger results, and provided a better assessment of the impact of isolation methods on intra-host viral diversity.
15
The categories of this paper are: developmental
22
The categories of this paper are: physiology
20
The categories of this paper are: pharmacology
13
The categories of this paper are: animal
21
The categories of this paper are: toxicology
1
The categories of this paper are: bioinformatics
32
The categories of this paper are: zoology
18
The categories of this paper are: systems
3
The categories of this paper are: evolutionary
30
The categories of this paper are: scientific
8
The categories of this paper are: genomics
2
The categories of this paper are: biology
31
The categories of this paper are: paleontology
17
The categories of this paper are: molecular
19
The categories of this paper are: epidemiology
14
The categories of this paper are: biophysics