title: Isolation of Infective Zika Virus from Urine and Saliva of Patients in Brazil.
abstract: Zika virus (ZIKV) is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications, prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. Here, we described the presence and isolation of infectious ZIKV particles from saliva and urine of acute phase patients in the Rio de Janeiro state, Brazil.
  
  
  
  Nine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak.
  
  
  
  The detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological relevance of this finding, regarding the contribution of alternative non-vectorial ZIKV transmission routes, needs further investigation.
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Myrna C
authors.identifier: 0000-0001-9899-3386
authors.initials: MC
authors.lastname: Bonaldo
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Ieda P
authors.initials: IP
authors.lastname: Ribeiro
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Noemia S
authors.initials: NS
authors.lastname: Lima
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Alexandre A C
authors.initials: AA
authors.lastname: Dos Santos
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Lidiane S R
authors.initials: LS
authors.lastname: Menezes
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Stephanie O D
authors.initials: SO
authors.lastname: da Cruz
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Iasmim S
authors.initials: IS
authors.lastname: de Mello
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Nathália D
authors.initials: ND
authors.lastname: Furtado
authors.affiliation: Laboratório de Biologia Molecular de Flavivírus, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Elaine E
authors.initials: EE
authors.lastname: de Moura
authors.affiliation: Laboratório de Doenças Febris Agudas, Instituto Nacional de Infectologia Evandro Chagas, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Luana
authors.initials: L
authors.lastname: Damasceno
authors.affiliation: Laboratório de Mosquitos Transmissores de Hematozoários, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Kely A B
authors.initials: KA
authors.lastname: da Silva
authors.affiliation: Laboratório de Mosquitos Transmissores de Hematozoários, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Marcia G
authors.initials: MG
authors.lastname: de Castro
authors.affiliation: Laboratório Nacional de Computação Científica, Petrópolis, Rio de Janeiro, Brazil.
authors.forename: Alexandra L
authors.initials: AL
authors.lastname: Gerber
authors.affiliation: Laboratório Nacional de Computação Científica, Petrópolis, Rio de Janeiro, Brazil.
authors.forename: Luiz G P
authors.initials: LG
authors.lastname: de Almeida
authors.affiliation: Laboratório de Mosquitos Transmissores de Hematozoários, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Lourenço-de-Oliveira
authors.affiliation: Laboratório Nacional de Computação Científica, Petrópolis, Rio de Janeiro, Brazil.
authors.forename: Ana Tereza R
authors.initials: AT
authors.lastname: Vasconcelos
authors.affiliation: Laboratório de Doenças Febris Agudas, Instituto Nacional de Infectologia Evandro Chagas, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Brasil
mesh_terms: D000328:Adult; D000368:Aged; D001938:Brazil / Q000453:epidemiology; D005260:Female; D016679:Genome, Viral; D006801:Humans; D008875:Middle Aged; D010802:Phylogeny; D011247:Pregnancy; D012367:RNA, Viral / Q000145:classification / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012463:Saliva / Q000821:virology*; D055815:Young Adult; D000071244:Zika Virus / Q000302:isolation & purification*; D000071243:Zika Virus Infection / Q000175:diagnosis* / Q000453:epidemiology / Q000652:urine*
chemical_list: D012367:RNA, Viral