title: An efficient in vivo method for the isolation of Puumala virus in Syrian hamsters and the characterization of the isolates from Russia.
abstract: Puumala virus (PUUV) and other Arvicolinae-borne hantaviruses are difficult to cultivate in cell culture. To isolate these hantaviruses efficiently, hantavirus nucleocapsid protein (NP)-positive but seronegative wild rodents were selected by NP-detection ELISA. Three of 68 Myodes glareolus captured in Samara, Russia, were NP-positive and seronegative. Syrian hamsters were inoculated with lung homogenates from NP-positive rodents for virus propagation. Virus isolation in vitro was carried out by inoculation of lung homogenates of NP-positive hamsters to Vero E6 cell monolayers. Two PUUV strains (Samara49/CG/2005 and Samara94/CG/2005) from M. glareolus were isolated in Vero E6 cells. Nucleotide and amino acid sequence identities of the S segment of these isolates to those of PUUV F-s808 from a fatal HFRS patient in Samara region were 96.7-99.3% and 99.3-100.0%, respectively. Morphologic features of Vero E6 cells infected with PUUV strain Samara49/CG/2005 were quite similar to those of Hantaan virus-infected cells. Isolation of Hokkaido virus from Myodes rufocanus captured in Hokkaido, Japan, was also performed. Hokkaido virus NP and RNA were recovered and maintained in hamsters. These results suggest that inoculation of Syrian hamsters with rodent samples is an efficient method for the isolation and maintenance of PUUV and other Arvicolinae-borne hantaviruses.
authors.affiliation: Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Seto
authors.forename: Evgeniy A
authors.initials: EA
authors.lastname: Tkachenko
authors.forename: Vyacheslav G
authors.initials: VG
authors.lastname: Morozov
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Tanikawa
authors.forename: Sergey I
authors.initials: SI
authors.lastname: Kolominov
authors.forename: Sergey N
authors.initials: SN
authors.lastname: Belov
authors.forename: Ichiro
authors.initials: I
authors.lastname: Nakamura
authors.forename: Nobuo
authors.initials: N
authors.lastname: Hashimoto
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kon
authors.forename: Alexander E
authors.initials: AE
authors.lastname: Balakiev
authors.forename: Tamara K
authors.initials: TK
authors.lastname: Dzagurnova
authors.forename: Olga A
authors.initials: OA
authors.lastname: Medvedkina
authors.forename: Mina
authors.initials: M
authors.lastname: Nakauchi
authors.forename: Mariko
authors.initials: M
authors.lastname: Ishizuka
authors.forename: Kentaro
authors.initials: K
authors.lastname: Yoshii
authors.forename: Kumiko
authors.initials: K
authors.lastname: Yoshimatsu
authors.forename: Leonid V
authors.initials: LV
authors.lastname: Ivanov
authors.forename: Jiro
authors.initials: J
authors.lastname: Arikawa
authors.forename: Ikuo
authors.initials: I
authors.lastname: Takashima
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kariwa
mesh_terms: D000818:Animals; D003411:Arvicolinae / Q000821:virology*; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007564:Japan; D008647:Mesocricetus; D023421:Models, Animal; D008969:Molecular Sequence Data; D029262:Puumala virus / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics; D012426:Russia; D017422:Sequence Analysis, DNA; D014709:Vero Cells; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation
chemical_list: D012367:RNA, Viral