• Title, Summary, Keyword: reovirus type 3

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Isolation and Characterization of Reovirus in Korea (한국에 분포하는 레오바이러스의 분리 및 동정)

  • Song, Ki-Joon;Kang, Byung-Chul;Lee, Young-Eun;Baek, Luck-Ju;Lee, Yong-Ju;Song, Jin-Won
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.65-74
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    • 1999
  • Reovirus was found to inhabit both the respiratory and the enteric tract of human and animals. The genome of reovirus comprises 10 segments of double-stranded RNA, total size 24 kbp. Nine strains of reovirus were isolated from human and field mice in Korea. Aseptically collected sera from human and lung tissues from field mice were used for virus isolation. For serotype determination, hemagglutination inhibition test was used, and three strains were confirmed to type 2 and six strains to type 3. To determine the genomic diversity and molecular phylogeny of reoviruses isolated in Korea, part of S4 genomic segment of reovirus was enzymatically amplified and directly sequenced. In nucleotide level, Apo98-35 strain showed 15.4%, 19.3%, and 14.4% differences compared to type 1 (T1L, Lang), type 2 (T2J), and type 3 reference strains, respectively. In amino acid level, Apo98-35 strain showed 10.5%, 13.7%, and 9.5% differences compared to type 1, type 2, and type 3 reference strains, respectively. Using the maximum parsimony method based on 285 bp spaning region of the S4 genomic segment, phylogenetic analysis indicated that Apo98-35 from Korea formed different phylogenetic branch. Our data obtained by sequence and phylogenetic analyses of reoviruses are consistent with the distinct geographically dependent evolution of reoviruses in Korea.

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Studies on Properties of Avian Reovirus Isolated in Korea (국내에서 분리한 닭 레오바이러스의 성상에 관한 연구)

  • 김성식;박병옥;김순재
    • Korean Journal of Veterinary Service
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    • v.15 no.1
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    • pp.67-80
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    • 1992
  • Avian reoviruses have been implicated in respiratory disease enteric conditions including Cloacal pasting in young thicks, pericarditis, hydropericardium, anaemia with swollen spleen and liver and petechiation of skeletal muscle and viral arthritis. This study was conducted to examine properties of reovirus field 3 strains isolated from affected broiler from several farms. An infectious agent was isolated from leg tendons and intestine of broiler with clinical tenosynovitis. The agent grew well on the chicken embryo kideny cells(CEK). One of them produced cytopathic effects(CPE) of round type and formed intranuclear inclusions, and the other was characterized by CPE of syncytical type and cytoplasmic inclusion. The properties and serological classification of field strains were examined by hemagglutin test, virus neutralization test, agar gel precipitin, electropherotype. They showed no hemagglutination reactions and not well neutralization and to possess common antigens detectable by AGP test. RNA electropherotype presented 10 segment band as the previous report. These data suggest that the field strains and standard strains (1133, 1733) may be the same group.

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A Study of the HI Antibody of the Koreans and Swine to Reovirus (한국인 및 가축(돼지)에 있어 Reovirus에 대한 HI 항체분석)

  • Lee, Yun-Tai;Lee, Chong-Hoon
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.77-84
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    • 1980
  • The purpose of this paper is to study the incidence of humoral antibody to reovirus type 2 in the sera of the Koreans and animal(swine) at random. All the 614 of human beings and 877 of swine sera were collected during the period from June to December, 1979, from the healthy persons in the National Seoul Hospital and swine blood was collected from 25 different areas of June to 30th of September in 1977. The HI test was put with plastic plates according to the methods by Rosen(1960 a and 1974). The total 73.29% of the 614 cases in human and the 61.80% of the 877 in swine confirmed as a hemagglutination inhibition antibodies. In human the 76.47% of the 442 male cases and the 65.12% of the 172 female ones were confirmed to have humoral antibodies. The positive rate was widely shown in each age group. But the 31 to 50 old age groups showed a little higher than any other age group, which came to the 85.71% in 41-50 and the 78.72% in 31-40 old age groups. The monthly distribution of HI antibody was shown to reach the 93.22% of the 59 cases in June. This per cent was much higher than of any other distribution. Accordingly, the auther came to the conclusion that there is reovirus type 2 in all the parts of Korea and most of the Koreans have the higher rates of antibody. However, the positive rate of antibody was the 542 out of the 877 cases(61.8%) from the swine and antibodies was confirmed from the 25 different areas in Korea. The seasonal distribution of the antibody showed these high rates. In domestics animals; blood from the swine showed that distribution of HI antibodies to reoviras type 2. These antibody appears from the various areas of the province in Korea. For this reasons, reovirus was widely distributed in this country.

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Real-Time RT-PCR for Validation of Reovirus Type 3 Safety During the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Reovirus Type 3 안전성 검증을 위한 Real-Time RT-PCR)

  • Lee, Dong-Hyuck;Jeong, Hyo-Sun;Kim, Tae-Eun;Oh, Seon-Hwan;Lee, Jung-Suk;Kim, In-Seop
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.228-236
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to Reovirus type 3 (Reo-3), and there are several reports of Reo-3 contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the Reo-3 safety, a real-time RT-PCR method was developed for quantitative detection of Reo-3 in cell lines, raw materials, manufacturing processes, and final products as well as Reo-3 clearance validation. Specific primers for amplification of Reo-3 RNA was selected, and Reo-3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $3.2{\times}10^0\;TCID_{50}/ml$. The real-time RT-PCR method was proven to be reproducible and very specific to Reo-3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with Reo-3. Reo-3 RNA could be quantified in CHO cell as well as culture supernatant. When the real-time RT-PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of Reo-3.

Evaluation of Viral Inactivation Efficacy of a Continuous Flow Ultraviolet-C Reactor (UVivatec) (연속 유동 Ultraviolet-C 반응기(UVivatec)의 바이러스 불활화 효과 평가)

  • Bae, Jung-Eun;Jeong, Eun-Kyo;Lee, Jae-Il;Lee, Jeong-Im;Kim, In-Seop;Kim, Jong-Su
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.377-382
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    • 2009
  • Viral safety is an important prerequisite for clinical preparations of all biopharmaceuticals derived from plasma, cell lines, or tissues of human or animal origin. To ensure the safety, implementation of multiple viral clearance (inactivation and/or removal) steps has been highly recommended for manufacturing of biopharmaceuticals. Of the possible viral clearance strategies, Ultraviolet-C (UVC) irradiation has been known as an effective viral inactivating method. However it has been dismissed by biopharmaceutical industry as a result of the potential for protein damage and the difficulty in delivering uniform doses. Recently a continuous flow UVC reactor (UVivatec) was developed to provide highly efficient mixing and maximize virus exposure to the UV light. In order to investigate the effectiveness of UVivatec to inactivate viruses without causing significant protein damage, the feasibility of the UVC irradiation process was studied with a commercial therapeutic protein. Recovery yield in the optimized condition of $3,000\;J/m^2$ irradiation was more than 98%. The efficacy and robustness of the UVC reactor was evaluated with regard to the inactivation of human immunodeficiency virus (HIV), hepatitis A virus (HAV), bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), porcine parvovirus (PPV), bovine parvovirus (BPV), minute virus of mice (MVM), reovirus type 3 (REO), and bovine parainfluenza virus type 3 (BPIV). Non enveloped viruses (HAV, PPV, BPV, MVM, and REO) were completely inactivated to undetectable levels by $3,000\;J/m^2$ irradiation. Enveloped viruses such as HIV, BVDV, and BPIV were completely inactivated to undetectable levels. However BHV was incompletely inactivated with slight residual infectivity remaining even after $3,000\;J/m^2$ irradiation. The log reduction factors achieved by UVC irradiation were ${\geq}3.89$ for HIV, ${\geq}5.27$ for HAV, 5.29 for BHV, ${\geq}5.96$ for BVDV, ${\geq}4.37$ for PPV, ${\geq}3.55$ for BPV, ${\geq}3.51$ for MVM, ${\geq}4.20$ for REO, and ${\geq}4.15$ for BPIV. These results indicate that UVC irradiation using UVivatec was very effective and robust in inactivating all the viruses tested.