{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T22:58:03Z","timestamp":1767913083165,"version":"3.49.0"},"reference-count":39,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2013,9,24]],"date-time":"2013-09-24T00:00:00Z","timestamp":1379980800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>A West Nile Virus (WNV) lineage 2 strain, named Nea Santa-Greece-2010, has been demonstrated to be responsible for the large outbreaks of neuroinvasive disease (WNND) that have been occurring in Greece since 2010, based on sequence similarities of viral isolates identified between 2010\u20132012. However, knowledge on the evolution of this strain is scarce because only partial WNV genome sequences are available from Greece. The aim of this study was to get the complete genome sequence of WNV from patients with infection. To this aim, plasma and urine samples collected during the 2012 Greek outbreak were retrospectively investigated. Full WNV genome sequence was obtained from a patient with WNND. The genome had 99.7% sequence identity to Nea Santa, higher than to other related WNV lineage 2 strains, and five amino acid changes apparently not relevant for viral pathogenicity or fitness. In addition, infection by WNV lineage 2 was confirmed in additional nine patients with WNND; in three of them the infection with WNV Nea Santa was demonstrated by sequencing. In conclusion, this study characterized for the first time a WNV full genome from a patient with WNND from Greece, demonstrated the persistence of the Nea Santa strain, and suggested that the virus might have locally evolved.<\/jats:p>","DOI":"10.3390\/v5092311","type":"journal-article","created":{"date-parts":[[2013,9,24]],"date-time":"2013-09-24T13:38:52Z","timestamp":1380029932000},"page":"2311-2319","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Genome Sequencing of West Nile Virus from Human Cases in Greece, 2012"],"prefix":"10.3390","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0720-8456","authenticated-orcid":false,"given":"Luisa","family":"Barzon","sequence":"first","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"},{"name":"Microbiology and Virology Unit, Padova University Hospital, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna","family":"Papa","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Medical School, Aristotle University of Thessaloniki,  Thessaloniki 54621, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monia","family":"Pacenti","sequence":"additional","affiliation":[{"name":"Microbiology and Virology Unit, Padova University Hospital, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisa","family":"Franchin","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"},{"name":"Microbiology and Virology Unit, Padova University Hospital, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3902-8356","authenticated-orcid":false,"given":"Enrico","family":"Lavezzo","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Squarzon","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giulia","family":"Masi","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Martello","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Theodolinta","family":"Testa","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Medical School, Aristotle University of Thessaloniki,  Thessaloniki 54621, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Riccardo","family":"Cusinato","sequence":"additional","affiliation":[{"name":"Microbiology and Virology Unit, Padova University Hospital, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giorgio","family":"Pal\u00f9","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"},{"name":"Microbiology and Virology Unit, Padova University Hospital, 35122 Padova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2013,9,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1159\/000328320","article-title":"West Nile virus: The complex biology of an emerging pathogen","volume":"54","author":"Ulbert","year":"2011","journal-title":"Intervirology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1007\/s10096-003-1085-1","article-title":"West Nile Virus: An overview of its spread in Europe and the Mediterranean basin in contrast to its spread in the Americas","volume":"23","author":"Zeller","year":"2004","journal-title":"Eur. J. Clin. Microbiol. Infect.Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1016\/S0140-6736(98)03538-7","article-title":"West Nile encephalitis epidemic in southeastern Romania","volume":"352","author":"Tsai","year":"1998","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"373","DOI":"10.3201\/eid0604.000408","article-title":"Isolation of two strains of West Nile virus during an outbreak in southern Russia, 1999","volume":"6","author":"Lvov","year":"2000","journal-title":"Emerg. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"128","DOI":"10.3201\/eid0701.010118","article-title":"Outbreak of West Nile virus infection, Volgograd Region, Russia, 1999","volume":"7","author":"Platonov","year":"2001","journal-title":"Emerg. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2333","DOI":"10.1126\/science.286.5448.2333","article-title":"Origin of the West Nile virus responsible for an outbreak of encephalitis in the northeastern United States","volume":"286","author":"Lanciotti","year":"1999","journal-title":"Science"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1001\/jama.2013.8042","article-title":"West Nile virus: Review of the literature","volume":"310","author":"Petersen","year":"2013","journal-title":"JAMA"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1111\/1469-0691.12211","article-title":"West Nile virus in Europe: emergence, epidemiology, diagnosis, treatment, and prevention","volume":"19","author":"Sambri","year":"2013","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_9","unstructured":"Hellenic Centre for Disease Control and Prevention (HCDCP): West Nile virus Revised Epidemic Report. Available online:http:\/\/www.keelpno.gr\/."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1868","DOI":"10.3201\/eid1710.110525","article-title":"Outbreak of West Nile virus infection in Greece, 2010","volume":"17","author":"Danis","year":"2011","journal-title":"Emerg. Infect. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Papa, A. (2013). West Nile virus infections in humans-Focus on Greece. J. Clin. Virol.","DOI":"10.1016\/j.jcv.2013.02.020"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1681","DOI":"10.1111\/j.1469-0691.2011.03642.x","article-title":"Non-neuroinvasive West Nile virus infections during the outbreak in Greece","volume":"17","author":"Anastasiadou","year":"2011","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Danis, K., Papa, A., Papanikolaou, E., Dougas, G., Terzaki, I, Baka, A., Vrioni, G., Kapsimali, V., Tsakris, A., and Kansouzidou, A. (2011). Ongoing outbreak of West Nile virus infection in humans, Greece, July to August 2011. Euro Surveill., 16.","DOI":"10.2807\/ese.16.34.19951-en"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"920","DOI":"10.3201\/eid1705.101759","article-title":"Genetic characterization of West Nile virus lineage 2, Greece, 2010","volume":"17","author":"Papa","year":"2011","journal-title":"Emerg. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"618","DOI":"10.3201\/eid1204.051379","article-title":"Lineage 1 and 2 strains of encephalitic West Nile virus, central Europe","volume":"12","author":"Bakonyi","year":"2006","journal-title":"Emerg. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"222","DOI":"10.3201\/eid1402.070457","article-title":"Genetic determinants of virulence in pathogenic lineage 2 West Nile virus strains","volume":"14","author":"Botha","year":"2008","journal-title":"Emerg. Infect. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1089\/vbz.2009.0230","article-title":"West Nile virus lineage 2 as a cause of zoonotic neurological disease in human and horses in southern Africa","volume":"10","author":"Venter","year":"2010","journal-title":"Vector Borne Zoonotic Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1176","DOI":"10.1111\/j.1469-0691.2010.03438.x","article-title":"Detection of West Nile virus lineage 2 in mosquitoes during a human outbreak in Greece","volume":"17","author":"Papa","year":"2011","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Papa, A., Papadopoulou, E., Gavana, E., Kalaitzopoulou, S., and Mourelatos, S. (2013). Detection of West Nile virus lineage 2 in Culex mosquitoes, Greece, 2012. Vector Borne Zoonotic Dis.","DOI":"10.1089\/vbz.2012.1212"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Chaskopoulou, A., Dovas, C.I., Chaintoutis, S.C., Kashefi, J., Koehler, P., and Papanastassopoulou, M. (2013). Detection and early warning of West Nile cirus circulation in Central Macedonia, Greece, using sentinel chickens and mosquitoes. Vector Borne Zoonotic Dis.","DOI":"10.1089\/vbz.2012.1176"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1551","DOI":"10.1007\/s00436-013-3302-x","article-title":"West Nile virus in mosquitoes in Greece","volume":"112","author":"Papa","year":"2013","journal-title":"Parasitol. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"684","DOI":"10.3201\/eid1904.121594","article-title":"West Nile virus infection in Belgian traveler returning from Greece","volume":"19","author":"Cnops","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1186\/1743-422X-9-266","article-title":"Serological and molecular investigation into the role of wild birds in the epidemiology of West Nile virus in Greece","volume":"9","author":"Valiakos","year":"2012","journal-title":"Virol. J."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Valiakos, G., Touloudi, A., Iacovakis, C., Athanasiou, L., Birtsas, P., Spyrou, V., and Billinis, C. (2011). Molecular detection and phylogenetic analysis of West Nile virus lineage 2 in sedentary wild birds (Eurasian magpie), Greece, 2010. Euro Surveill., 16.","DOI":"10.2807\/ese.16.18.19862-en"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Chaskopoulou, A., Dovas, C., Chaintoutis, S., Bouzalas, I., Ara, G., and Papanastassopoulou, M. (2011). Evidence of enzootic circulation of West Nile virus (Nea Santa-Greece-2010, lineage 2), Greece, May to July 2011. Euro Surveill., 16.","DOI":"10.2807\/ese.16.31.19933-en"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"827","DOI":"10.3201\/eid1905.121418","article-title":"West Nile virus lineage 2 strain in Greece, 2012","volume":"19","author":"Chaintoutis","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.meegid.2013.03.034","article-title":"Epidemiological history and phylogeography of West Nile virus lineage 2","volume":"17","author":"Ciccozzi","year":"2013","journal-title":"Infect. Genet. Evol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1099\/vir.0.046888-0","article-title":"Molecular evolution of lineage 2 West Nile virus","volume":"94","author":"McMullen","year":"2013","journal-title":"J. Gen. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1162","DOI":"10.1038\/ng2097","article-title":"A single positively selected West Nile viral mutation confers increased virogenesis in American crows","volume":"39","author":"Brault","year":"2007","journal-title":"Nat. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1111\/j.1365-3148.2009.00964.x","article-title":"Searching for West Nile virus (WNV) in Greece","volume":"20","author":"Kantzanou","year":"2010","journal-title":"Transfus Med"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Dimou, V., Gerou, S., and Papa, A. (2013). The epidemic West Nile virus strain in Greece was a recent introduction. Vector Borne Zoonotic Dis.","DOI":"10.1089\/vbz.2013.1318"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/BF01731581","article-title":"A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences","volume":"16","author":"Kimura","year":"1980","journal-title":"J. Mol. Evol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"783","DOI":"10.2307\/2408678","article-title":"Confidence limits on phylogenies: An approach using the bootstrap","volume":"39","author":"Felsenstein","year":"1985","journal-title":"Evolution"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2731","DOI":"10.1093\/molbev\/msr121","article-title":"MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods","volume":"28","author":"Tamura","year":"2011","journal-title":"Mol. Biol. Evol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Barzon, L., Pacenti, M., Franchin, E., Pagni, S., Martello, T., Cattai, M., Cusinato, R., and Pal\u00f9, G. (2013). Excretion of West Nile virus in urine during acute infection. J. Infect. Dis.","DOI":"10.1093\/infdis\/jit290"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4066","DOI":"10.1128\/JCM.38.11.4066-4071.2000","article-title":"Rapid detection of West Nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay","volume":"38","author":"Lanciotti","year":"2000","journal-title":"J. Clin. Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1016\/j.jviromet.2007.05.021","article-title":"Detection of West Nile virus lineages 1 and 2 by real-time PCR","volume":"146","author":"Linke","year":"2007","journal-title":"J. Virol. Methods."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1922","DOI":"10.1128\/JCM.39.5.1922-1927.2001","article-title":"Comparison of flavivirus universal primer pairs and development of a rapid, highly sensitive heminested reverse transcription-PCR assay for detection of flaviviruses targeted to a conserved region of the NS5 gene sequences","volume":"39","author":"Scaramozzino","year":"2001","journal-title":"J.Clin. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Barzon, L., Pacenti, M., Cusinato, R., Cattai, M., Franchin, E., Pagni, S., Martello, T., Bressan, S., Squarzon, L., and Cattelan, A. (2011). Human cases of West Nile Virus Infection in north-eastern Italy, 15 June to 15 November 2010. Euro Surveill., 16.","DOI":"10.2807\/ese.16.33.19949-en"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/5\/9\/2311\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:49:28Z","timestamp":1760219368000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/5\/9\/2311"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,9,24]]},"references-count":39,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2013,9]]}},"alternative-id":["v5092311"],"URL":"https:\/\/doi.org\/10.3390\/v5092311","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,9,24]]}}}