title: An efficient method for isolation of Hantaan virus through serial passages in suckling mice.
abstract: OBJECTIVE
  Hantaan virus (HTNV) is one of the main etiologic agents for hemorrhagic fever with renal syndrome in China. However, it is very difficult to isolate the virus from its original host, which hampers the viral characterization. This study describes an efficient method for isolating HTNV in suckling mice.
  
  
  METHODS
  Hantavirus-infected Apodemus agrarius were screened by quantitative real-time PCR. The homogenates of one positive rodent lung tissue were inoculated into suckling mice for virus propagation through serial passages.
  
  
  RESULTS
  During the three passages in suckling mice, the number of viral RNA copies/nanogram of GAPDH mRNA increased significantly ranging from 477 to 7,278 and 46 to 4,898 in the tissues of brain and lung, respectively. Hantaviral antigens could be detected by indirect immunofluorescence assay and around 100-nm virion-like structures were also observed in brain tissue by transmission electron microscopy. No nucleotide exchange was found except for one in the 3'-non-coding domain of S segment when comparing the complete genome sequences from hantavirus in the first and the third passages.
  
  
  CONCLUSIONS
  These results suggest inoculation of suckling mice with suspected hantavirus-infected rodent samples is an efficient method for isolation and maintenance of HTNV.
authors.affiliation: State Key Laboratory of Virology, Institute of Medical Virology, School of Medicine, Wuhan University, Wuhan, PR China.
authors.forename: J-L
authors.initials: JL
authors.lastname: Li
authors.forename: J-X
authors.initials: JX
authors.lastname: Ling
authors.forename: L-J
authors.initials: LJ
authors.lastname: Chen
authors.forename: F
authors.initials: F
authors.lastname: Wei
authors.forename: F
authors.initials: F
authors.lastname: Luo
authors.forename: Y-Y
authors.initials: YY
authors.lastname: Liu
authors.forename: H-R
authors.initials: HR
authors.lastname: Xiong
authors.forename: W
authors.initials: W
authors.lastname: How
authors.forename: Z-Q
authors.initials: ZQ
authors.lastname: Yang
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D018063:Hantaan virus / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D046529:Microscopy, Electron, Transmission; D051189:Murinae / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012692:Serial Passage / Q000379:methods; D014771:Virion / Q000648:ultrastructure; D014773:Virology / Q000379:methods*
chemical_list: D012367:RNA, Viral