G. MoureauS. TemmamJ. P. GonzalezR. N. CharrelG. GrardX. De LamballerieFaculte de Medecine de Marseille Universite de la MediterraneeMahidol University2018-08-242018-08-242007-12-01Vector-Borne and Zoonotic Diseases. Vol.7, No.4 (2007), 467-477153036672-s2.0-37549000515https://repository.li.mahidol.ac.th/handle/20.500.14594/24476Here we describe an optimized molecular protocol for the universal detection and identification of flaviviruses. It combines the convenient real-time polymerase chain reaction (PCR) format with a broad spectrum of flavivirus detection. This assay, based on the amplification of a 269-272 nt (depending on the flavivirus tested) region at the N terminal end of the NS5 gene, enabled the amplification of 51 flavivirus species and 3 tentative species. Sequencing of the amplicons produced by reverse transcriptase (RT)-PCR permitted the reliable taxonomic identification of flavivirus species by comparison with reference sequences available in databases, using either the BLASTN algorithm or a simple phylogenetic reconstruction. The limit of detection of the assay (2-20,500 copies/reaction depending on the virus tested) allowed the detection of different flaviviruses from a series of human sera or veterinary samples. Altogether, the characteristics of this technique make it a good candidate for the identification of previously identified flaviviruses in cell culture and the investigation of field samples, and also a promising tool for the discovery and identification of new species, including viruses distantly related to "classical" arthropod-borne flaviviruses. © 2007 Mary Ann Liebert, Inc.Mahidol UniversityImmunology and MicrobiologyMedicineArticleSCOPUS10.1089/vbz.2007.0206