• Title/Summary/Keyword: Hantaan isolation

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Correlation of Virologic Property and Phylogenetic Analysis of Hantaan Viruses Isolated from Patients and Reservoirs in Korea

  • Jung, Yong-Tae;Lee, Sun-Ryang;Paik, Soon-Young;Kim, Gum-Ryong
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.11-20
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    • 1998
  • Twenty isolates of Hantavirus were isolated from patients and reserovirs from 1988 to 1994 in Korea. Isolation rate was 1.9% (10/538) in patients, 6.2% (5/81) in Apodemus sp., 2.6% (1/38) in Rattus sp. and 0.6% (4/677) in bats. Reciprocal mean IFA titers ranged from 27.5 to 1,024 at the specimen collection. According to the growth rate and reaching peak titier of infectivity, the isolates were grouped as rapid, intermediate, and slow growing groups. All isolates were confirmed as Hantaan type by the nested RT-PCR on the G1 region of the M segment. Comparison of nucleotide sequence (Nt: 2101 - Nt: 2280) of the G2 region revealed that the sequence homology bewteen Hantaan 76/118 virus and the isolates was more than 90%. Several nucleotide positions of the isolates showed high variation. The variation rate of patientisolates was about one-half when compared with that of rodentisolates. On the basis of phylogenetic analysis Hantaan viruses isolated were divided into two genogroups. These results indicate that Hantaan virus is highly dominant serotype in Korea and the virologic property and genogroup are not correlated.

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Hantavirus infection and isolation from wild shrews(Crocidura lasiura) in Korea (국내 야생따쥐(Crocidura lasiura) 의 Hantavirus 항체 보유실태 및 원인체 분리)

  • Kim, Hee-sun;Kang, Mun-il
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.147-152
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    • 1994
  • Eleven shrews were caught from three areas of Korea. All of them were confirmed in the same species, Crocidura lasiura. All of sera from wild shrews were examined by indirect immunofluorescent test against Hantaan-related virus. The antibody to Hantaan-related virus was detected by 2 out of 11 shrews. Just 2 of 7 shrews from BG area were sero-positive for Hantaan-related virus antigen and none from other. All of sero-positive for Hantaan-related virus antigen belonged to male with antibody titer of 1:40 to 1:80. Two Hantaan-related viruses were isolated in vivo and in vitro.

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Isolation and Purification of Hantaan Viral Nucleocapsid Protein Expressed in Escherichia coli (대장균에서 발현된 한탄바이러스 뉴클레오캡시드 단백질의 분리 정제)

  • 노갑수;김종완;하석훈;정근택;문상범;최차용
    • KSBB Journal
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    • v.13 no.6
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    • pp.656-661
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    • 1998
  • Hantaan virus belonging to the genus Hantavirus and family Bunyaviridae causes an acute severe illness of human, Haemorrhagic Fever with Renal Syndrome (HFRS). It is a rodent host-borne pathogen and distributed in Asia and Eastern Europe. Hantaviruses have three major antigens, i.e., G1, G2 glycoproteins and nucleocapsid protein (N). Among them, nucleocapsid protein was reported to be the most invaluable antigen as for diagnosis. We have cloned and expressed Hantaan viral nucleocapsid gene in E. coli BL21(DE3). In this study, we have tried to purify the nucleocapsid protein produced by recombinant E. coli, and could attained a purity of >90% by anti-N monoclonal antibody-coupled immunoaffinity chromatography or phenyl sepharose hydrophobic interaction chromatography.

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Passage and Adaptation of Maaji Virus in Hamster (Maaji Virus의 Hamster 계대 및 적응)

  • Kim, Yun-Cheol;Paik, Woo-Hyun;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.67-76
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    • 1996
  • The methods that make Hantavirus grow consist of inoculation into the experimental animals and cultured cells. The cultured cells, such as Vero-E6 and A549 cells, have been usually used for isolation of the virus and the animals, such as mice and rats, are used for large scale preparation of the virus so far. Furthermore, the cell can be used to maintain the virus and assay the infectivity and the animals can be used for the experiment of viral pathogenicity and challenge for assessment of vaccine. Apodemus mice, the own natural host of the virus, has been used for challenge test of Hantaan virus. However it has been pointed out to difficult handling and breeding the animal in laboratory. Therefore, we attempted to establish a new animal model for challenge test at the time of isolation of Maaji virus which is a new hantavirus similar but distinct to Hantaan virus. In suckling hamster, the titer of Maaji virus and the lethality to mice of the virus were increased gradually in the titer and lethality through passage by intracerebral (IC) inoculation. We tried to re-adapt this brain virus to lung of weanling hamster. The brain passaged virus was inoculated into weanling hamster intramuscularly. Again, the titer of the virus in lung was also increased by continuous passage of this virus. This facts could regarded as adaptation to new environment in which the virus proliferates. To identity the virus passaged in hamster with Maaji virus, both of the virus passaged in hamster brain and lung were compared with Maaji virus (MAA-I) and Hantaan virus (HTN 76-118) by means of restriction fragment length polymorphism (RFLP) and slingle strand conformation polymophism (SSCP). As a result, we conclude that Maaji virus could be adapted successfully to weanling hamster through this passage strategy. Utilizing this adapted Maaji virus strain, hamster model is able to be used for challenge test in hantaviral vaccinology and further experiments utilizing hamster system as a rather available and convenient lab animal are expected.

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Isolation and Genetic Study of Hantavirus from Apodemus peninsulae Captured in Yeuncheon-gun, Kyunggi-do (경기도에서 채집한 Apodemus peninsulae에서 한탄바이러스 분리와 유전학적 연구)

  • Song, Ki-Joon;Kim, Yong-Soo;Lee, Yong-Ju;Kang, Ju-Il;Song, Jin-Won;Baek, Luck-Ju
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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    • pp.337-345
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    • 1998
  • Hantaviruses are distributed in rodent population world-widely even in geographical areas where hemorrhagic fever with renal syndrome (HFRS) has not been reported. Various species of Family Muridae and Arvicolidae serve as the natural reservoirs of hantaviruses. Hantaan virus, Seoul virus, Puumala virus, Prospect HII virus, Sin Nombre virus and New York virus are members of genus Hantavirus and isolated from lungs of A. agrarius, R. norvegicus, C. glareolus, M. pennsylvanicus, P. maniculatus and P. leucopus respectively. This experiment was intended to find the distribution of hantavirus infection among wild rodents and isolate the hantavirus from lung tissue of seropositve Apodemus peninsulae, and compared the nucleotide and amino acid sequences with prototype of hantaan virus 76-118 strain. Hantaviral sequences were amplified from lung tissues of A. peninsulae by reverse-transcriptase polymerase chain reaction. Alignment and comparison of the 324 nucleotide of G2 region of M-genomic segment diverged 4.6% and 0% at the nucleotide and amino acid levels, and complete N protein-coding region of S-genomic segment diverged 3.7% and 1.4% nucleotide and amino acid levels, respectively. This is the report to spill-over on the hantaan virus from A. agrarius to A. peninsulae in Korea.

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Isolation of the Hantaviruses from the Lungs of Bandicota indica Captured in Indonesia and Thailand (인도네시아와 태국에서 채집된 Bandicota indica 폐장조직에서 한타바이러스 분리)

  • Woo, Young-Dae;Chu, Yong-Kyu;Lee, Do-Wang
    • The Journal of Korean Society of Virology
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    • v.28 no.2
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    • pp.157-164
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    • 1998
  • Various hantaviruses were isolated from HFRS patients and various rodent species, in many parts of the world. Bandicotas were captured at Yogyakarta, east region of Sumatura island, Indonesia; and 4 rodents species including Bandicotas were captured at Chiang Rai in Thailand during 1995. Sera were collected from captured bandicotas and other rodent spicies were screened for antibody test against Hantaan (HTN), Seoul (SEO), Puumala (PUU) and Sin Nombre (SN) viruses by immunofluoresence antibody assay (IFA). Hantavirus antigen in lung tissues were tested by IFA. Among 55 captured Bandicota indica in Indonesia, 14 (25.5%) were antibody positive against HTN, SEO, PUU and SN virus. Hantavirus antigen were detected from 5 (9.0%) out of 55 lungs tested. Among 34 captured Bandicota indica in Thailand, 9 (26.5%) were antibody positive against HTN, SEO, PUU and SN virus. Among 34 lungs tissues of Bandicota indica examined, 3 (8.8%) were antigen positive. In other rodent species, antibody positive against Hantaviruses of Rattus rattus, Rattus losea and Mus cervicolor were 4/62 (6.5%), 5/25 (20%), 1/1 (100%), respectively. But no one has antigen in their lung tissues. Antigen positive lungs suspension were inoculated into vero E6 cells for virus isolation and 4 viruses were isolated from Indonesian Badicota and 3 viruses from Thailand.

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Isolation of Apodemus peninsulae-borne Hantavirus and Comparison of Molecular Biological Characteristics (흰넓적다리붉은쥐 유래 한타바이러스 분리 및 분자생물학적 특성 비교)

  • Song, Ki-Joon;Yun, Hyung-Seon;Kho, Eun-Young;Chung, Ki-Mo;Park, Kwang-Sook;Lee, Yong-Ju;Song, Jin-Won;Baek, Luck-Ju
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.19-28
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    • 2000
  • Two distinct hantaviruses have been isolated from Apodemus agrarius in 1976 and Rattus norvegicus in 1980 in Korea. Since our serosurveys conducted in 1994, a genetically distinct hantavirus from Apodemus peninsulae has been investigated. To isolate hantavirus from Apen insulae captured in Korea, the lung homogenate of seropositive Apeninsulae inoculated Vero E6 cells. Viral antigen was detected in a progressively higher percentage of cells with subsequent passage after 80 days postinoculation. The new isolate from seropositive Apodemus peninsulae was designated Suchong virus after Suchong valley located in northeastern region of South Korea. Comparing with hantaan virus 76-118 strain, Suchong virus-1, 2, 3 and 4 showed the similarity of $71.0{\sim}91.8%$ at nucleotide and $90.9{\sim}94.8%$ at amino acid sequences in 231 nucleotides region of M segment, and the similarity of $75.1{\sim}81.0%$ at nucleotide and $97.5{\sim}100%$ at amino acid sequences in 237 nucleotides of S segment.

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