title: Complete genome sequencing of dengue virus type 1 isolated in Buenos Aires, Argentina.
abstract: Dengue (DEN) constitutes a major viral arthropod-borne human illness. South America was last considered free of dengue two decades ago when a dramatic increase in the number of dengue fever and hemorrhagic dengue cases had been reported. Five viruses were isolated in Buenos Aires City from the 1999-2000 Paraguay outbreak. RT-PCRs obtained directly from plasma were cloned into pGemT vectors and sequences of the structural genes and NS1 were analyzed. Three viruses were full-length sequenced from RT-PCR obtained from cell-culture isolates. Excess of synonymous over non-synonymous mutations suggested that the structural proteins were under strong functional constraints while a weak purifying selection was operating in the whole polyprotein. Sequence diversity and selective pressures varied among patients but results were significantly above the procedure threshold. One sample showed small-plaque phenotype and impaired growth coupled to 3'untranslated region mutations. Phylogenetic analysis of full-length sequences split Buenos Aires isolates into two clusters within American DEN-1 genotype V: Clade I was phylogenetically linked to Brazilian samples and Clade II with samples from Paraguay and Northeastern Argentina. In Buenos Aires City, only dengue virus serotype 1 imported from Paraguay has been detected, though without evidence of local transmission.
authors.affiliation: Laboratorio de Virología, Hospital de Niños Dr. Ricardo Gutiérrez., Gallo 1330, 1425 Buenos Aires, Argentina. paobarrero@hotmail.com
authors.forename: Paola R
authors.initials: PR
authors.lastname: Barrero
authors.forename: Alicia S
authors.initials: AS
authors.lastname: Mistchenko
mesh_terms: D020413:3' Untranslated Regions; D001118:Argentina; D002460:Cell Line; D003716:Dengue Virus / Q000145:classification / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification*; D005814:Genes, Viral; D014644:Genetic Variation; D016679:Genome, Viral; D023281:Genomics; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D012367:RNA, Viral / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D017361:Viral Nonstructural Proteins / Q000235:genetics; D010948:Viral Plaque Assay; D015678:Viral Structural Proteins / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; C082515:NS1 protein, dengue-1 virus; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins