An efficient in vivo method for the isolation of Puumala virus in Syrian hamsters and the characterization of the isolates from Russia

Seto, Takeshi; Ткаченко, Е. А.; Морозов, В. Г.; Tanikawa, Yoichi; Kolominov, Sergey I.; Belov, Sergey N.; Nakamura, Ichiro; Hashimoto, Nobuo; Kon, Yasuhiro; Balakiev, Alexander E.; Dzagurnova, Tamara K.; Medvedkina, Olga A.; Nakauchi, Mina; Ishizuka, Mariko; Yoshii, Kentaro; Yoshimatsu, Kumiko; Иванов, Л. И.; Arikawa, Jiro; Takashima, Ikuo; Kariwa, Hiroaki

doi:10.1016/j.jviromet.2010.12.019

Cited by 9 publications

(10 citation statements)

References 31 publications

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“…For taxonomic identification, mtDNA was extracted from the liver of royal voles, using the High Pure Template Preparation Kit (Roche, Indianapolis, IN, USA). Nucleotide and amino acid sequences of each segment of MUJV were compared with PUUV [11,14,32], HOKV [6,33], LUXV [34], Prospect Hill virus [35], KBRV [27], TULV [36], SNV [37], HTNV [38], DOBV [39], Soochong virus [40], SEOV [41], and previously sequenced MUJV strains [8]. …”

Section: Methodsmentioning

confidence: 99%

Muju Virus, Harbored by Myodes regulus in Korea, Might Represent a Genetic Variant of Puumala Virus, the Prototype Arvicolid Rodent-Borne Hantavirus

Lee

Baek

et al. 2014

Viruses

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The genome of Muju virus (MUJV), identified originally in the royal vole (Myodes regulus) in Korea, was fully sequenced to ascertain its genetic and phylogenetic relationship with Puumala virus (PUUV), harbored by the bank vole (My. glareolus), and a PUUV-like virus, named Hokkaido virus (HOKV), in the grey red-backed vole (My. rufocanus) in Japan. Whole genome sequence analysis of the 6544-nucleotide large (L), 3652-nucleotide medium (M) and 1831-nucleotide small (S) segments of MUJV, as well as the amino acid sequences of their gene products, indicated that MUJV strains from different capture sites might represent genetic variants of PUUV, the prototype arvicolid rodent-borne hantavirus in Europe. Distinct geographic-specific clustering of MUJV was found in different provinces in Korea, and phylogenetic analyses revealed that MUJV and HOKV share a common ancestry with PUUV. A better understanding of the taxonomic classification and pathogenic potential of MUJV must await its isolation in cell culture.

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Section: Methodsmentioning

confidence: 99%

Muju Virus, Harbored by Myodes regulus in Korea, Might Represent a Genetic Variant of Puumala Virus, the Prototype Arvicolid Rodent-Borne Hantavirus

Lee

Baek

et al. 2014

Viruses

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“…Virus isolation. Virus isolation was performed using a modified version of a method described previously (Seto et al, 2011). Briefly, lung tissues from an HOKV-infected grey red-backed vole were inoculated into suckling Syrian hamsters intracerebrally.…”

Section: Methodsmentioning

confidence: 99%

Isolation of Hokkaido virus, genus Hantavirus, using a newly established cell line derived from the kidney of the grey red-backed vole (Myodes rufocanus bedfordiae)

Sanada

Seto

Ozaki

et al. 2012

Journal of General Virology

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Hantaviruses belong to the family Bunyaviridae and are maintained in wild rodents. Although Vero E6 cells, which originate from African green monkey kidney, are used widely in hantavirus research, isolation of hantaviruses from this cell line is difficult. To develop an efficient method of propagation and isolation of hantaviruses we established a novel cell line, MRK101, derived from the kidney of the grey red-backed vole (Myodes rufocanus bedfordiae), the natural host of Hokkaido virus (HOKV). The MRK101 cells showed a significantly higher susceptibility to Puumala virus (PUUV) hosted by Myodes glareolus than Vero E6 cells. Viral nucleocapsid protein in PUUV-infected MRK101 cells was detected earlier than in Vero E6 cells, and the viral titre in the culture fluid of MRK101 cells was higher than that of Vero E6 cells during the early phase of infection. In contrast, MRK101 cells showed no susceptibility to Hantaan virus. HOKV, which has not been isolated to date, was isolated successfully using MRK101 cells. Moreover, the newly isolated HOKV was successfully propagated in MRK101, but not Vero E6, cells. Phylogenic analyses of the S (small), M (medium) and L (large) segment sequences revealed that HOKV is related most closely to PUUV, but is distinct from other hantaviruses. These data suggest that the MRK101 cell line is a useful tool for the isolation and propagation of hantaviruses. Moreover, this is (to our knowledge) the first report of hantavirus isolation in a cell line that originated from the natural host. INTRODUCTIONHantaviruses, members of the family Bunyaviridae, have a tri-segmented, negative-stranded RNA genome consisting of small (S), medium (M) and large (L) segments (Plyusnin et al., 1996), which encode nucleocapsid (N) proteins, envelope glycoproteins (Gn and Gc) and an RNA-dependent RNA polymerase (RdRP), respectively (Plyusnin et al., 1996). Each hantavirus is carried by a specific species in the orders Rodentia and Soricomorpha. For example, Hantaan virus (HTNV), Seoul virus (SEOV), Amur virus (AMRV) and Puumala virus (PUUV) are carried by the striped field mouse (Apodemus agrarius), Norway rat (Rattus norvegicus), Korean field mouse (Apodemus peninsulae) and bank vole (Myodes glareolus), respectively (Jonsson et al., 2010). In general, hantaviruses are believed to infect persistently, and are apathogenic to their reservoir hosts (Gavrilovskaya et al., 1990;Yanagihara et al., 1985). In contrast, hantaviruses when transmitted to humans cause two severe disease syndromes: haemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). HTNV, SEOV, AMRV, Dobrava-Belgrade virus, Saaremaa virus and PUUV have been identified as aetiological agents of HFRS. Sin Nombre virus (SNV), Laguna Negra virus and Andes virus (ANDV) are considered to be the major pathogens associated with HCPS (Bi et al., 2008;Jonsson et al., 2010).Hokkaido virus (HOKV) is a hantavirus that is maintained in grey red-backed voles (Myodes rufocanus bedfordiae), which are distributed geographically ...

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“…Therefore, it may be that BALB/c mouse is more suitable for inoculation and isolation of HTNV than hantaviruses derived from voles. A recent study indicated that inoculation of Syrian hamsters with rodent samples was an efficient method for the isolation and maintenance of Puumala virus [26]. However, hamsters are more distantly related to Myodes glareolus (the natural host of Puumala virus), which is more likely to lead to the low susceptibility for the infection [24].…”

Section: Discussionmentioning

confidence: 99%

An Efficient Method for Isolation of Hantaan Virus through Serial Passages in Suckling Mice

Ling²,

Chen³

et al. 2013

Intervirology

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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. Conclusion: These results suggest inoculation of suckling mice with suspected hantavirus-infected rodent samples is an efficient method for isolation and maintenance of HTNV.

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An efficient in vivo method for the isolation of Puumala virus in Syrian hamsters and the characterization of the isolates from Russia

Cited by 9 publications

References 31 publications

Muju Virus, Harbored by Myodes regulus in Korea, Might Represent a Genetic Variant of Puumala Virus, the Prototype Arvicolid Rodent-Borne Hantavirus

Muju Virus, Harbored by Myodes regulus in Korea, Might Represent a Genetic Variant of Puumala Virus, the Prototype Arvicolid Rodent-Borne Hantavirus

Isolation of Hokkaido virus, genus Hantavirus, using a newly established cell line derived from the kidney of the grey red-backed vole (Myodes rufocanus bedfordiae)

An Efficient Method for Isolation of Hantaan Virus through Serial Passages in Suckling Mice

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