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Título
Real time PCR assay for detection of all known lineages of West Nile virus
Autor(es)
Vazquez, Ana ISCIII | Herrero-Romero, Laura ISCIII | Negredo, Anabel ISCIII | Hernandez, Lourdes ISCIII | Sánchez-Seco, María Paz ISCIII | Tenorio, Antonio ISCIII
Fecha de publicación
2016
Cita
J Virol Methods. 2016 Oct;236:266-270.
Idioma
Inglés
Tipo de documento
journal article
Resumen
West Nile virus (WNV) is one of the most widespread arbovirus and a large variety of WNV strains and lineages have been described. The molecular methods for the diagnosis of WNV target mainly lineages 1 and 2, which have caused outbreaks in humans, equines and birds. But the last few years new and putative WNV lineages of unknown pathogenicity have been described. Here we describe a new sensitive and specific real-time PCR assay for the detection and quantification of all the WNV lineages described until now. Primers and probe were designed in the 3'-untranslated region (3'-UTR) of the WNV genome and were designed to match all sequenced WNV strains perfectly. The sensitivity of the assay ranged from 1,5 to 15 copies per reaction depending on the WNV lineage tested. The method was validated for WNV diagnosis using different viral strains, human samples (cerebrospinal fluid, biopsies, serum and plasma) and mosquito pools. The assay did not amplify any other phylogenetically or symptomatically related viruses. All of the above make it a very suitable tool for the diagnosis of WNV and for surveillance studies.
Palabras clave
MESH
3' Untranslated Regions | Animals | Culicidae | DNA Primers | Horses | Humans | Oligonucleotide Probes | RNA, Viral | Real-Time Polymerase Chain Reaction | Sensitivity and Specificity | West Nile Fever | West Nile virus | Genotype
Versión en línea
DOI
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