title: Rapid molecular detection and genotyping of West Nile Virus lineages 1 and 2 by real time PCR and melting curve analysis.
abstract: Following its spread in the USA, West Nile Virus (WNV) has reemerged in the Mediterranean basin with a renewed pathogenicity. The introduction of WNV lineage 2 in Europe and its co-circulation with lineage 1 has resulted in a continuously changing epidemiological scenario, highlighting the importance of differential detection of the two lineages. The paper describes a new real-time PCR method for the detection and genotyping of the two main lineages of WNV. The method requires a single pair of primers and probes and is based on the analysis of highly conserved consensus sequences detected in the 5' terminus of the viral genome.
authors.affiliation: Laboratory of Molecular Biology, Army Medical Research Center, Via Santo Stefano Rotondo 4, 00184 Rome, Italy. Electronic address: giovanni.faggioni@gmail.com.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Faggioni
authors.affiliation: Laboratory of Molecular Biology, Army Medical Research Center, Via Santo Stefano Rotondo 4, 00184 Rome, Italy. Electronic address: riccardo.desantis@gmail.com.
authors.forename: Riccardo
authors.initials: R
authors.lastname: De Santis
authors.affiliation: Laboratory of Molecular Biology, Army Medical Research Center, Via Santo Stefano Rotondo 4, 00184 Rome, Italy. Electronic address: alice.pomponi@gmail.com.
authors.forename: Alice
authors.initials: A
authors.lastname: Pomponi
authors.affiliation: Department of Clinical Sciences and Translational Medicine, University of Rome "Tor Vergata", Rome, Italy. Electronic address: max.fantini@libero.it.
authors.forename: Massimo
authors.initials: M
authors.lastname: Fantini
authors.affiliation: Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Teramo, Italy. Electronic address: g.savini@izs.it.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Savini
authors.affiliation: Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Teramo, Italy. Electronic address: f.monaco@izs.it.
authors.forename: Federica
authors.initials: F
authors.lastname: Monaco
authors.affiliation: Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Teramo, Italy. Electronic address: a.polci@izs.it.
authors.forename: Andrea
authors.initials: A
authors.lastname: Polci
authors.affiliation: Department of Clinical Sciences and Translational Medicine, University of Rome "Tor Vergata", Rome, Italy. Electronic address: bei@med.uniroma2.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Bei
authors.affiliation: Laboratory of Molecular Biology, Army Medical Research Center, Via Santo Stefano Rotondo 4, 00184 Rome, Italy. Electronic address: romano.lista@gmail.com.
authors.forename: Florigio
authors.initials: F
authors.lastname: Lista
mesh_terms: D017931:DNA Primers / Q000235:genetics; D005060:Europe / Q000453:epidemiology; D016679:Genome, Viral; D060005:Genotyping Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D044366:Transition Temperature; D014481:United States / Q000453:epidemiology; D014901:West Nile Fever / Q000453:epidemiology / Q000821:virology*; D014902:West Nile virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D015345:Oligonucleotide Probes; D012367:RNA, Viral
keywords: Flavivirus; Melting curve analysis; Molecular detection; Virus genotyping; WNV