{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T22:31:07Z","timestamp":1780698667339,"version":"3.54.1"},"reference-count":27,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,16]],"date-time":"2021-03-16T00:00:00Z","timestamp":1615852800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>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 the 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. In this study, we describe the different stages of 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. High-throughput and Sanger sequencing results provided a better assessment of the impact of isolation methods on viral diversity.<\/jats:p>","DOI":"10.3390\/pathogens10030349","type":"journal-article","created":{"date-parts":[[2021,3,16]],"date-time":"2021-03-16T12:01:02Z","timestamp":1615896062000},"page":"349","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Isolation and Genetic Characterization of Puumala Orthohantavirus Strains from France"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3680-0088","authenticated-orcid":false,"given":"Johann","family":"Vulin","sequence":"first","affiliation":[{"name":"Laboratoire de Lyon, ANSES, Unit\u00e9 de Virologie, 69007 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0141-557X","authenticated-orcid":false,"given":"S\u00e9verine","family":"Murri","sequence":"additional","affiliation":[{"name":"Laboratoire de Lyon, ANSES, Unit\u00e9 de Virologie, 69007 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Madri\u00e8res","sequence":"additional","affiliation":[{"name":"Laboratoire de Lyon, ANSES, Unit\u00e9 de Virologie, 69007 Lyon, France"},{"name":"CBGP, INRAE, CIRAD, IRD, Institut Agro, Univ Montpellier, 34000 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6981-5732","authenticated-orcid":false,"given":"Maxime","family":"Galan","sequence":"additional","affiliation":[{"name":"CBGP, INRAE, CIRAD, IRD, Institut Agro, Univ Montpellier, 34000 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caroline","family":"Tatard","sequence":"additional","affiliation":[{"name":"CBGP, INRAE, CIRAD, IRD, Institut Agro, Univ Montpellier, 34000 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7717-7555","authenticated-orcid":false,"given":"Sylvain","family":"Piry","sequence":"additional","affiliation":[{"name":"CBGP, INRAE, CIRAD, IRD, Institut Agro, Univ Montpellier, 34000 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0215-3521","authenticated-orcid":false,"given":"Gabriele","family":"Vaccari","sequence":"additional","affiliation":[{"name":"Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanit\u00e0, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"D\u2019Agostino","sequence":"additional","affiliation":[{"name":"Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanit\u00e0, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7907-6539","authenticated-orcid":false,"given":"Nathalie","family":"Charbonnel","sequence":"additional","affiliation":[{"name":"CBGP, INRAE, CIRAD, IRD, Institut Agro, Univ Montpellier, 34000 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillaume","family":"Castel","sequence":"additional","affiliation":[{"name":"CBGP, INRAE, CIRAD, IRD, Institut Agro, Univ Montpellier, 34000 Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philippe","family":"Marianneau","sequence":"additional","affiliation":[{"name":"Laboratoire de Lyon, ANSES, Unit\u00e9 de Virologie, 69007 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.virusres.2013.12.019","article-title":"Discovery of hantaviruses and of the Hantavirus genus: Personal and historical perspectives of the Presidents of the International Society of Hantaviruses","volume":"187","author":"Lee","year":"2014","journal-title":"Virus Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1093\/infdis\/141.2.131","article-title":"Nephropathia Epidemica: Detection of Antigen in Bank Voles and Serologic Diagnosis of Human Infection","volume":"141","author":"Vaheri","year":"1980","journal-title":"J. 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