title: Urinary detection of toscana virus nucleic acids in neuroinvasive infections.
abstract: BACKGROUND
  Toscana virus (TOSV) is a sandfly-borne pathogen causing febrile diseases and neuroinvasive infections in humans. Definitive diagnosis of TOSV infections frequently requires the detection of viral RNA in cerebrospinal fluid (CSF) or in circulation, which can be achieved prior to seroconversion.
  
  
  OBJECTIVE
  To evaluate TOSV excretion in urine and impact of urine as a diagnostic specimen.
  
  
  METHODS
  A total of 82 plasma, CSF and urine samples were collected from 24 individuals with a preliminary diagnosis of atypical viral encephalitis, where frequent bacterial fungal and viral causes were ruled out. Phlebovirus and WNV nucleic acids were investigated via real-time and nested polymerase chain reaction (PCR) assays. Commercial immunofluorescence assays were employed for viral IgM detection. Amplicons were characterized via cloning and sequencing.
  
  
  RESULTS
  Phlebovirus PCR yielded positive results in 7 out of 14 samples that comprise 4 plasma and 3 urine specimens from 3 individuals. Amplicons were characterized as TOSV genotype A. Investigation of the follow-up samples suggested that virus shedding in urine coincides or follows viremia. Despite conserved sequences observed in paired or sequential plasma-urine specimens, L693S substitution in the viral polymerase was characterized in a urine sample.
  
  
  CONCLUSIONS
  These preliminary findings indicate that urine can be employed as a additional clinical sample for TOSV RNA detection in suspected cases, especially in individuals where specimens for viral diagnostics during the early stages of the infection are not available.
authors: Hacettepe University, Faculty of Medicine, Department of Medical Microbiology, Virology Unit, Morphology Building 3rd floor, 06100 Sihhiye, Ankara, Turkey. Electronic address: ekoray@hacettepe.edu.tr.
authors: Koray
authors: K
authors: Ergunay
authors: Hacettepe University, Faculty of Medicine, Department of Neurology, 06100 Ankara, Turkey.
authors: Berkan
authors: B
authors: Kaplan
authors: Hacettepe University, Faculty of Medicine, Department of Neurology, 06100 Ankara, Turkey.
authors: Serhat
authors: S
authors: Okar
authors: Ankara University, Faculty of Veterinary Medicine, Department of Virology, 06110 Ankara, Turkey.
authors: Zeynep
authors: Z
authors: Akkutay-Yoldar
authors: Hacettepe University, Faculty of Medicine, Department of Neurology, 06100 Ankara, Turkey.
authors: Asli
authors: A
authors: Kurne
authors: Hacettepe University, Faculty of Medicine, Department of Neurology, 06100 Ankara, Turkey.
authors: Ethem Murat
authors: EM
authors: Arsava
authors: Ankara University, Faculty of Veterinary Medicine, Department of Virology, 06110 Ankara, Turkey.
authors: Aykut
authors: A
authors: Ozkul
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009422:Nervous System Diseases / Q000175:diagnosis* / Q000821:virology*; D010217:Phlebotomus Fever / Q000175:diagnosis* / Q000821:virology*; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics*; D029301:Sandfly fever Naples virus / Q000145:classification / Q000235:genetics*; D016415:Sequence Alignment
chemical_list: D012367:RNA, Viral
keywords: Diagnosis; Excretion; Phlebovirus; Toscana virus; Urine