{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T14:54:20Z","timestamp":1780325660035,"version":"3.54.1"},"reference-count":24,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2014,4,4]],"date-time":"2014-04-04T00:00:00Z","timestamp":1396569600000},"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>The determination of levels of rabies virus-neutralizing antibody (VNA) provides the foundation for the quantitative evaluation of immunity effects. The traditional fluorescent antibody virus neutralization test (FAVN) using a challenge virus standard (CVS)-11 strain as a detection antigen and staining infected cells with a fluorescein isothiocyanate (FITC)-labeled monoclonal antibody, is expensive and high-quality reagents are often difficult to obtain in developing countries. Indeed, it is essential to establish a rapid, economical, and specific rabies virus neutralization test (VNT). Here, we describe a recombinant virus rCVS-11-eGFP strain that stably expresses enhanced green fluorescent protein (eGFP) based on a reverse genetic system of the CVS-11 strain. Compared to the rCVS-11 strain, the rCVS-11-eGFP strain showed a similar growth property with passaging stability in vitro and pathogenicity in vivo. The rCVS-11-eGFP strain was utilized as a detection antigen to determine the levels of rabies VNAs in 23 human and 29 canine sera; this technique was termed the FAVN-eGFP method. The good reproducibility of FAVN-eGFP was tested with partial serum samples. Neutralization titers obtained from FAVN and FAVN-eGFP were not significantly different. The FAVN-eGFP method allows rapid economical, specific, and high-throughput assessment for the titration of rabies VNAs.<\/jats:p>","DOI":"10.3390\/v6041578","type":"journal-article","created":{"date-parts":[[2014,4,4]],"date-time":"2014-04-04T11:11:12Z","timestamp":1396609872000},"page":"1578-1589","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Generation of Recombinant Rabies Virus CVS-11 Expressing eGFP Applied to the Rapid Virus Neutralization Test"],"prefix":"10.3390","volume":"6","author":[{"given":"Xianghong","family":"Xue","sequence":"first","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"},{"name":"College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuexing","family":"Zheng","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"},{"name":"Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongru","family":"Liang","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Na","family":"Feng","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongkun","family":"Zhao","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuwei","family":"Gao","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hualei","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Songtao","family":"Yang","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"},{"name":"College of Veterinary Medicine, Jilin University, Changchun 130062, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianzhu","family":"Xia","sequence":"additional","affiliation":[{"name":"Institute of Military Veterinary Medicine, Academy of Military Medical Science,  Changchun 130122, China"},{"name":"College of Veterinary Medicine, Jilin University, Changchun 130062, China"},{"name":"Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2014,4,4]]},"reference":[{"key":"ref_1","first-page":"45","article-title":"Pathogenesis of rabies","volume":"292","author":"Dietzschold","year":"2005","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1186\/1743-422X-9-159","article-title":"Infection with street strain rabies virus induces modulation of the microRNA profile of the mouse brain","volume":"9","author":"Zhao","year":"2012","journal-title":"Virol. J."},{"key":"ref_3","unstructured":"World Health Organization (WHO) (1999). The World Survey for Rabies No. 35 Geneva Document WHO\/CDS\/CSR\/EPH\/2002.10, WHO."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1089\/vbz.2008.0046","article-title":"Rabies in China: An update","volume":"9","author":"Hu","year":"2009","journal-title":"Vector-Borne Zoonotic Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.vetmic.2006.04.009","article-title":"Canine vaccination\u2014Providing broader benefits for disease control","volume":"117","author":"Cleaveland","year":"2006","journal-title":"Vet. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1017\/S0950268802006957","article-title":"Comparison of vaccination strategies for the control of dog rabies in Machakos District, Kenya","volume":"129","author":"Kitala","year":"2002","journal-title":"Epidemiol. Infect."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1371\/journal.pbio.1000053","article-title":"Transmission dynamics and prospects for the elimination of Canine Rabies","volume":"7","author":"Hampson","year":"2009","journal-title":"PLoS Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/0264-410X(95)00197-9","article-title":"Immunization coverage required to prevent outbreaks of dog rabies","volume":"14","author":"Coleman","year":"1996","journal-title":"Vaccine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3200","DOI":"10.1128\/JVI.06699-11","article-title":"Faber M.Postexposure treatment with the live-attenuated rabies virus (RV) vaccine TriGAS triggers the clearance of wild-type RV from the Central Nervous System (CNS) through the rapid induction of genes relevant to adaptive immunity in CNS tissues","volume":"86","author":"Li","year":"2012","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1186\/1423-0127-19-61","article-title":"Sufficient virus-neutralizing antibody in the central nerve system improves the survival of rabid rats","volume":"19","author":"Liao","year":"2012","journal-title":"J. Biomed. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"218","DOI":"10.17221\/5827-VETMED","article-title":"Comparison of the detection and quantification of rabies antibodies in canine sera","volume":"47","author":"Ondrejkova","year":"2002","journal-title":"Vet. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1006\/biol.1998.0162","article-title":"A Comparison of two serological methods for detecting the immune response after rabies vaccination in dogs and cats being exported to rabies-free areas","volume":"26","author":"Briggs","year":"1998","journal-title":"Biologicals"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/s12250-013-3321-x","article-title":"Establishment and preliminary application of a rapid fluorescent focus inhibition test (RFFIT) for rabies virus","volume":"28","author":"Yu","year":"2013","journal-title":"Virol. Sin."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.virusres.2007.07.018","article-title":"Glycoprotein gene relocation in rabies virus","volume":"131","author":"Wu","year":"2008","journal-title":"Virus Res."},{"key":"ref_15","first-page":"43","article-title":"Rescue of recombinant peste des petits ruminants virus: Creation of a GFP-expressing virus and application in rapid virus neutralization test","volume":"43","author":"Wu","year":"2012","journal-title":"Vet. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.virusres.2009.12.009","article-title":"Development of a reverse genetics system for a human rabies virus vaccine strain employed in China","volume":"149","author":"Huang","year":"2010","journal-title":"Virus Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1309","DOI":"10.1128\/JVI.76.3.1309-1327.2002","article-title":"Relative neurotropism of a recombinant rhabdovirus expressing a green fluorescent envelope glycoprotein","volume":"76","author":"Dalton","year":"2002","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"12333","DOI":"10.1128\/JVI.78.22.12333-12343.2004","article-title":"Tracking fluorescence-labeled rabies virus: Enhanced green fluorescent protein-tagged phosphoprotein P supports virus gene expression and formation of infectious particles","volume":"78","author":"Finke","year":"2004","journal-title":"J. Virol."},{"key":"ref_19","first-page":"409","article-title":"Sequencing the complete genome of rabies virus CVS-11 strain and constructing its full-length infectious cDNA clone","volume":"4","author":"Xue","year":"2013","journal-title":"Acta Microbiol. Sin."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/S0166-0934(02)00249-5","article-title":"An improved method for recovering rabies virus from cloned cDNA","volume":"107","author":"Inoue","year":"2003","journal-title":"J. Virol. Methods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/S0022-1759(97)00212-3","article-title":"Development of a fluorescent antibody virus neutralisation test (FAVN test) for the quantification of rabies neutralising antibody","volume":"212","author":"Sagne","year":"1998","journal-title":"J. Immunol. Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/S0166-3542(96)01207-7","article-title":"Photodynamic inactivation of enveloped viruses by buckminsterfullerene","volume":"34","year":"1997","journal-title":"Antivir. Res."},{"key":"ref_23","unstructured":"(2005). Statistical Program for Social Sciences (SPSS), SPSS Inc.. version 13.0."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.jviromet.2004.12.003","article-title":"A novel rapid fluorescent focus inhibition test for rabies virus using a recombinant rabies virus visualizing a green fluorescent protein","volume":"125","author":"Khawplod","year":"2005","journal-title":"J. Virol. 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