title: Evidence of West Nile virus infection in Nepal.
abstract: BACKGROUND
  Acute febrile illness is common among those seeking medical care and is frequently treated empirically with the underlying illness remaining undiagnosed in resource-poor countries. A febrile illness study was conducted 2009-2010 to identify known and unknown pathogens circulating in Nepal.
  
  
  METHODS
  Study methods included diagnostic testing and preliminary ELISA screening of acute and convalescent samples for diseases both known and unknown to be circulating in Nepal, including West Nile virus (WNV). The molecular assays including Polymerase Chain Reaction (PCR), Sanger sequencing and ultra deep sequencing on MiSeq Illumina Platform were conducted to further confirm the presence of WNV.
  
  
  RESULTS
  The study enrolled 2,046 patients presenting undifferentiated febrile illness with unknown etiology. Sera from 14 out of 2,046 patients were tested positive for west nile virus (WNV) by nested Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Only two out of 14 cases were confirmed for the presence of WNV by sequencing and identified as WNV lineage 1 phylogentically. The two patients were adult males with fever and no neurological symptoms from Kathmandu and Bharatpur, Nepal.
  
  
  CONCLUSIONS
  Two out of 2,046 serum samples contained fragments of WNV genome resembling WNV lineage 1, which is evidence of the continued spread of WNV which should be considered a possible illness cause in Nepal.
authors.affiliation: Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand. wiriyar@afrims.org.
authors.forename: Wiriya
authors.initials: W
authors.lastname: Rutvisuttinunt
authors.affiliation: Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand. PiyawanC@afrims.org.
authors.forename: Piyawan
authors.initials: P
authors.lastname: Chinnawirotpisan
authors.affiliation: Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand. ChontichaK@afrims.org.
authors.forename: Chonticha
authors.initials: C
authors.lastname: Klungthong
authors.affiliation: Walter Reed/AFRIMS Research Unit Nepal, Kathmandu, Nepal. ShresthaSK@afrims.org.
authors.forename: Sanjaya Kumar
authors.initials: SK
authors.lastname: Shrestha
authors.affiliation: Bharatpur Hospital, Chitwan, Nepal. dramodbd_thapa@hotmail.com.
authors.forename: Amod Bahadur
authors.initials: AB
authors.lastname: Thapa
authors.affiliation: Sukra Raj Tropical Infectious Diseases Hospital, Kathmandu, Nepal. arpant20@yahoo.com.
authors.forename: Arjun
authors.initials: A
authors.lastname: Pant
authors.affiliation: Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand. YingstSL@afrims.org.
authors.forename: Samuel L
authors.initials: SL
authors.lastname: Yingst
authors.affiliation: Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand. YoonI@afrims.org.
authors.forename: In-Kyu
authors.initials: IK
authors.lastname: Yoon
authors.affiliation: Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand. Fernandezs@afrims.org.
authors.forename: Stefan
authors.initials: S
authors.lastname: Fernandez
authors.affiliation: Armed Forces Health Surveillance Center, Silver Spring, Maryland, USA. julie.a.pavlin.civ@mail.mil.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Pavlin
mesh_terms: D000328:Adult; D000368:Aged; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005334:Fever / Q000209:etiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D009390:Nepal / Q000453:epidemiology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D016415:Sequence Alignment; D014901:West Nile Fever / Q000150:complications / Q000453:epidemiology* / Q000821:virology; D014902:West Nile virus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D012367:RNA, Viral