{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T19:38:20Z","timestamp":1773085100292,"version":"3.50.1"},"reference-count":40,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2011,5,1]],"date-time":"2011-05-01T00:00:00Z","timestamp":1304208000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2011,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Rabies is a fatal zoonosis caused by a nonsegmented negative-strand RNA virus, namely, rabies virus (RABV). Apart from RABV, at least 10 additional species are known as rabies-related lyssaviruses (RRVs), and some of them are responsible for occasional spillovers into humans. More lyssaviruses have also been detected recently in different bat ecosystems, thanks to the application of molecular diagnostic methods. Due to the variety of the members of the genus\n            <jats:named-content content-type=\"genus-species\">Lyssavirus<\/jats:named-content>\n            , there is the necessity to develop a reliable molecular assay for rabies diagnosis able to detect and differentiate among the existing rabies and rabies-related viruses. In the present study, a pyrosequencing protocol targeting the 3\u2032 terminus of the nucleoprotein (N) gene was applied for the rapid characterization of lyssaviruses. Correct identification of species was achieved for each sample tested. Results from the pyrosequencing assay were also confirmed by those obtained using the Sanger sequencing method. A pan-lyssavirus one-step reverse transcription (RT)-PCR was developed within the framework of the pyrosequencing procedure. The sensitivity (Se) of the one-step RT-PCR assay was determined by using\n            <jats:named-content content-type=\"genus-species\">in vitro<\/jats:named-content>\n            -transcribed RNA and serial dilutions of titrated viruses. The assay demonstrated high analytical and relative specificity (Sp) (98.94%) and sensitivity (99.71%). To date, this is the first case in which pyrosequencing has been applied for lyssavirus identification using a cheaper diagnostic approach than the one for all the other protocols for rapid typing that we are acquainted with. Results from this study indicate that this procedure is suitable for lyssavirus detection in samples of both human and animal origin.\n          <\/jats:p>","DOI":"10.1128\/jcm.02015-10","type":"journal-article","created":{"date-parts":[[2011,3,10]],"date-time":"2011-03-10T03:53:13Z","timestamp":1299729193000},"page":"1932-1938","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":43,"title":["Lyssavirus Detection and Typing Using Pyrosequencing"],"prefix":"10.1128","volume":"49","author":[{"given":"Paola","family":"De Benedictis","sequence":"first","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cristian","family":"De Battisti","sequence":"additional","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laurent","family":"Dacheux","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Lyssavirus Dynamics and Host Adaptation Unit, National Reference Centre for Rabies, WHO Collaborating Centre for Reference and Research on Rabies, 28 rue du Docteur Roux, 75724 Paris, Cedex 15, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sabrina","family":"Marciano","sequence":"additional","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Silvia","family":"Ormelli","sequence":"additional","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela","family":"Salomoni","sequence":"additional","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Silvia Tiozzo","family":"Caenazzo","sequence":"additional","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anthony","family":"Lepelletier","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Lyssavirus Dynamics and Host Adaptation Unit, National Reference Centre for Rabies, WHO Collaborating Centre for Reference and Research on Rabies, 28 rue du Docteur Roux, 75724 Paris, Cedex 15, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Herv\u00e9","family":"Bourhy","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Lyssavirus Dynamics and Host Adaptation Unit, National Reference Centre for Rabies, WHO Collaborating Centre for Reference and Research on Rabies, 28 rue du Docteur Roux, 75724 Paris, Cedex 15, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ilaria","family":"Capua","sequence":"additional","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanni","family":"Cattoli","sequence":"additional","affiliation":[{"name":"OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Universit\u00e0 10, Legnaro, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","unstructured":"Anonymous. 2005. 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