{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T21:57:38Z","timestamp":1772143058815,"version":"3.50.1"},"reference-count":40,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2016,9,16]],"date-time":"2016-09-16T00:00:00Z","timestamp":1473984000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006454","name":"Federal Food Safety and Veterinary Office","doi-asserted-by":"publisher","award":["1.13.09"],"award-info":[{"award-number":["1.13.09"]}],"id":[{"id":"10.13039\/501100006454","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Rabies is a highly fatal zoonotic disease which is primarily caused by rabies virus (RABV) although other members of the genus Lyssavirus can cause rabies as well. As yet, 14 serologically and genetically diverse lyssaviruses have been identified, mostly in bats. To assess the quality of rabies vaccines and immunoglobulin preparations, virus neutralization tests with live RABV are performed in accordance with enhanced biosafety standards. In the present work, a novel neutralization test is presented which takes advantage of a modified vesicular stomatitis virus (VSV) from which the glycoprotein G gene has been deleted and replaced by reporter genes. This single-cycle virus was trans-complemented with RABV envelope glycoprotein. Neutralization of this pseudotype virus with RABV reference serum or immune sera from vaccinated mice showed a strong correlation with the rapid fluorescent focus inhibition test (RFFIT). Importantly, pseudotype viruses containing the envelope glycoproteins of other lyssaviruses were neutralized by reference serum to a significantly lesser extent or were not neutralized at all. Taken together, a pseudotype virus system has been successfully developed which allows the safe, fast, and sensitive detection of neutralizing antibodies directed against different lyssaviruses.<\/jats:p>","DOI":"10.3390\/v8090254","type":"journal-article","created":{"date-parts":[[2016,9,19]],"date-time":"2016-09-19T10:07:43Z","timestamp":1474279663000},"page":"254","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Quantification of Lyssavirus-Neutralizing Antibodies Using Vesicular Stomatitis Virus Pseudotype Particles"],"prefix":"10.3390","volume":"8","author":[{"given":"Sarah","family":"Moeschler","sequence":"first","affiliation":[{"name":"Institut f\u00fcr Virologie und Immunologie (IVI), Abteilung Virologie, CH-3147 Mittelh\u00e4usern, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samira","family":"Locher","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Virologie und Immunologie (IVI), Abteilung Virologie, CH-3147 Mittelh\u00e4usern, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karl-Klaus","family":"Conzelmann","sequence":"additional","affiliation":[{"name":"Max von Pettenkofer-Institut und Genzentrum, Ludwig-Maximilians-Universit\u00e4t, D-81377 M\u00fcnchen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Beate","family":"Kr\u00e4mer","sequence":"additional","affiliation":[{"name":"Paul-Ehrlich-Institut, Abteilung Veterin\u00e4rmedizin, D-63225 Langen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2708-2507","authenticated-orcid":false,"given":"Gert","family":"Zimmer","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Virologie und Immunologie (IVI), Abteilung Virologie, CH-3147 Mittelh\u00e4usern, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2016,9,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/S1473-3099(02)00287-6","article-title":"Rabies re-examined","volume":"2","author":"Rupprecht","year":"2002","journal-title":"Lancet Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2351","DOI":"10.1007\/s00705-016-2880-1","article-title":"Taxonomy of the order Mononegavirales: Update 2016","volume":"161","author":"Afonso","year":"2016","journal-title":"Arch. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3268","DOI":"10.1128\/JVI.75.7.3268-3276.2001","article-title":"Evidence of two Lyssavirus phylogroups with distinct pathogenicity and immunogenicity","volume":"75","author":"Badrane","year":"2001","journal-title":"J. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Delmas, O., Holmes, E.C., Talbi, C., Larrous, F., Dacheux, L., Bouchier, C., and Bourhy, H. (2008). Genomic diversity and evolution of the lyssaviruses. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0002057"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/j.meegid.2009.02.005","article-title":"The rhabdoviruses: Biodiversity, phylogenetics, and evolution","volume":"9","author":"Kuzmin","year":"2009","journal-title":"Infect. Genet. Evol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.virusres.2008.04.021","article-title":"Complete genomes of Aravan, Khujand, Irkut and West Caucasian bat viruses, with special attention to the polymerase gene and non-coding regions","volume":"136","author":"Kuzmin","year":"2008","journal-title":"Virus Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2799","DOI":"10.4161\/21645515.2014.972741","article-title":"Cross-neutralization of antibodies induced by vaccination with Purified Chick Embryo Cell Vaccine (PCECV) against different Lyssavirus species","volume":"10","author":"Malerczyk","year":"2014","journal-title":"Hum. Vaccin. Immunother."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1389","DOI":"10.1016\/S0140-6736(13)62707-5","article-title":"Current status of rabies and prospects for elimination","volume":"384","author":"Fooks","year":"2014","journal-title":"Lancet"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1002\/jmv.10481","article-title":"Case report: Isolation of a European bat lyssavirus type 2a from a fatal human case of rabies encephalitis","volume":"71","author":"Fooks","year":"2003","journal-title":"J. Med. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"959","DOI":"10.4269\/ajtmh.1972.21.959","article-title":"A fatal human infection with Mokola virus","volume":"21","author":"Familusi","year":"1972","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e1063","DOI":"10.1542\/peds.2013-1782","article-title":"Australian Bat Lyssavirus in a child: The first reported case","volume":"133","author":"Francis","year":"2014","journal-title":"Pediatrics"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.3201\/eid1212.060764","article-title":"Fatal human infection with rabies-related Duvenhage virus, South Africa","volume":"12","author":"Paweska","year":"2006","journal-title":"Emerg. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"481","DOI":"10.2217\/17460794.3.5.481","article-title":"Concepts in the pathogenesis of rabies","volume":"3","author":"Dietzschold","year":"2008","journal-title":"Future Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"754","DOI":"10.1128\/iai.16.3.754-759.1977","article-title":"Rabies virus glycoprotein. II. Biological and serological characterization","volume":"16","author":"Cox","year":"1977","journal-title":"Infect. Immun."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"7447","DOI":"10.1016\/j.vaccine.2012.10.015","article-title":"Rabies virus vaccines: Is there a need for a pan-lyssavirus vaccine?","volume":"30","author":"Evans","year":"2012","journal-title":"Vaccine"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Hampson, K., Coudeville, L., Lembo, T., Sambo, M., Kieffer, A., Attlan, M., Barrat, J., Blanton, J.D., Briggs, D.J., and Cleaveland, S. (2015). Estimating the global burden of endemic canine rabies. PLoS Negl. Trop. Dis., 9.","DOI":"10.1371\/journal.pntd.0003786"},{"key":"ref_17","first-page":"360","article-title":"Re-evaluating the burden of rabies in Africa and Asia","volume":"83","author":"Knobel","year":"2005","journal-title":"Bull. World Health Organ."},{"key":"ref_18","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 quantitation of rabies-neutralising antibody","volume":"212","author":"Cliquet","year":"1998","journal-title":"J. Immunol. Methods"},{"key":"ref_19","first-page":"535","article-title":"A rapid reproducible test for determining rabies neutralizing antibody","volume":"48","author":"Smith","year":"1973","journal-title":"Bull. World Health Organ."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Moore, S.M., and Hanlon, C.A. (2010). Rabies-specific antibodies: Measuring surrogates of protection against a fatal disease. PLoS Negl. Trop. Dis., 4.","DOI":"10.1371\/journal.pntd.0000595"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7178","DOI":"10.1016\/j.vaccine.2009.09.024","article-title":"A robust lentiviral pseudotype neutralisation assay for in-field serosurveillance of rabies and lyssaviruses in Africa","volume":"27","author":"Wright","year":"2009","journal-title":"Vaccine"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1099\/vir.0.80788-0","article-title":"Use of influenza C virus glycoprotein HEF for generation of vesicular stomatitis virus pseudotypes","volume":"86","author":"Hanika","year":"2005","journal-title":"J. Gen. Virol."},{"key":"ref_23","first-page":"219","article-title":"Chimeric rabies viruses for trans-species comparison of lyssavirus glycoprotein ectodomain functions in virus replication and pathogenesis","volume":"125","author":"Genz","year":"2012","journal-title":"Berl. Munchener Tierarztliche Wochenschr."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1016\/0042-6822(90)90433-R","article-title":"Molecular cloning and complete nucleotide sequence of the attenuated rabies virus SAD B19","volume":"175","author":"Conzelmann","year":"1990","journal-title":"Virology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1302","DOI":"10.1099\/vir.0.82692-0","article-title":"Comparative analysis of the full genome sequence of European bat lyssavirus type 1 and type 2 with other lyssaviruses and evidence for a conserved transcription termination and polyadenylation motif in the G-L 3\u2032 non-translated region","volume":"88","author":"Marston","year":"2007","journal-title":"J. Gen. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1444","DOI":"10.1128\/jvi.69.3.1444-1451.1995","article-title":"Mokola virus glycoprotein and chimeric proteins can replace rabies virus glycoprotein in the rescue of infectious defective rabies virus particles","volume":"69","author":"Mebatsion","year":"1995","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.1016\/j.vaccine.2008.12.019","article-title":"A recombinant vesicular stomatitis virus replicon vaccine protects chickens from highly pathogenic avian influenza virus (H7N1)","volume":"27","author":"Kalhoro","year":"2009","journal-title":"Vaccine"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Berger Rentsch, M., and Zimmer, G. (2011). A vesicular stomatitis virus replicon-based bioassay for the rapid and sensitive determination of multi-species type I interferon. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0025858"},{"key":"ref_29","first-page":"37","article-title":"Collaborative study for validation of a serological potency assay for rabies vaccine (inactivated) for veterinary use","volume":"2010","author":"Bruckner","year":"2010","journal-title":"Pharmeuropa Bio Sci. Notes"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.biologicals.2009.01.001","article-title":"The rapid fluorescent focus inhibition test is a suitable method for batch potency testing of inactivated rabies vaccines","volume":"37","author":"Schildger","year":"2009","journal-title":"Biologicals"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/2043-9113-2-5","article-title":"Monte Carlo simulation of the Spearman-Kaerber TCID50","volume":"2","author":"Wulff","year":"2012","journal-title":"J. Clin. Bioinform."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2782","DOI":"10.1099\/vir.0.023978-0","article-title":"Fusion-active glycoprotein G mediates the cytotoxicity of vesicular stomatitis virus M mutants lacking host shut-off activity","volume":"91","author":"Hoffmann","year":"2010","journal-title":"J. Gen. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.vetmic.2010.02.001","article-title":"Human rabies due to lyssavirus infection of bat origin","volume":"142","author":"Johnson","year":"2010","journal-title":"Vet. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.jviromet.2015.03.022","article-title":"A recombinant rabies virus expressing a phosphoprotein-eGFP fusion is rescued and applied to the rapid virus neutralization antibody assay","volume":"219","author":"Tang","year":"2015","journal-title":"J. Virol. Methods"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1578","DOI":"10.3390\/v6041578","article-title":"Generation of recombinant rabies Virus CVS-11 expressing eGFP applied to the rapid virus neutralization test","volume":"6","author":"Xue","year":"2014","journal-title":"Viruses"},{"key":"ref_36","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. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.virusres.2015.05.023","article-title":"G gene-deficient single-round rabies viruses for neuronal circuit analysis","volume":"216","author":"Ghanem","year":"2016","journal-title":"Virus Res."},{"key":"ref_38","first-page":"272","article-title":"Development and Applications of VSV Vectors Based on Cell Tropism","volume":"2","author":"Tani","year":"2011","journal-title":"Front. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1089\/vim.1992.5.213","article-title":"Conformation-dependent accessibility of the linear epitopes located on the rabies virus glycoprotein","volume":"5","author":"Kontsekova","year":"1992","journal-title":"Viral Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1111\/j.1348-0421.1998.tb02270.x","article-title":"Antigenic and functional analyses of glycoprotein of rabies virus using monoclonal antibodies","volume":"42","author":"Luo","year":"1998","journal-title":"Microbiol. Immunol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/8\/9\/254\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:31:06Z","timestamp":1760211066000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/8\/9\/254"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,9,16]]},"references-count":40,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2016,9]]}},"alternative-id":["v8090254"],"URL":"https:\/\/doi.org\/10.3390\/v8090254","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,9,16]]}}}