• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.3
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• pp.238-242
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• 2009

Recently coastal seawater has been contaminated by enteric viruses such as the norovirus via untreated groundwater globally. Accordingly, the consumption of molluscan shellfish from seawater that has been contaminated with fecal material has become an important issues. The levels of enteric viruses in seawater are low and recovery and concentration of the virus from large volumes of water is difficult. We compared the effectiveness of two representative method of concentrating virus using negatively and positively charged filters. The mean retention of seeded FCV by HAMF and NCCF was 48% and 78%, respectively. Overall, the recovery of NCCF was 43.3$\pm$11% better than that of HAMF. However, the eluate obtained by using beef extract solution in the NCCF procedure caused an inhibitory effect on the RT-PCR; therefore, it was necessary to employ a PCR inhibitor removal procedure. The HAMF eluate contained no PCR inhibitors, but HAMF was not an effective method of concentrating the virus from large volumes of natural seawater due to clogging.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.591-599
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• 2008

A murine model immunized by systemic and mucosal delivery of plasmid DNA vaccine expressing glycoprotein B (pCIgB) of pseudorabies virus (PrV) was used to evaluate both the nature of the induced immunity and protection against a virulent virus. With regard to systemic delivery, the intramuscular (i.m.) immunization with pCIgB induced strong PrV-specific IgG responses in serum but was inefficient in generating a mucosal IgA response. Mucosal delivery through intranasal (i.n.) immunization of pCIgB induced both systemic and mucosal immunity at the distal mucosal site. However, the levels of systemic immunity induced by i.n. immunization were less than those induced by i.m. immunization. Moreover, i.n. genetic transfer of pCIgB appeared to induce Th2-biased immunity compared with systemic delivery, as judged by the ratio of PrV-specific IgG isotypes and Th1- and Th2-type cytokines produced by stimulated T cells. Moreover, the immunity induced by i.n. immunization did not provide effective protection against i.n. challenge of a virulent PrV strain, whereas i.m. immunization produced resistance to viral infection. Therefore, although i.n. immunization was a useful route for inducing mucosal immunity at the virus entry site, i.n. immunization did not provide effective protection against the lethal infection of PrV.

• Microbiology and Biotechnology Letters
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• v.36 no.1
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• pp.12-20
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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 minute virus of mice(MVM), and there are several reports of MVM contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the MVM safety, a real-time PCR method was developed for quantitative detection of MVM in cell lines, raw materials, manufacturing processes, and final products as well as MVM clearance validation. Specific primers for amplification of MVM DNA was selected, and MVM DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $6{\times}10^{-2}TCID_{50}/mL$. The real-time PCR method was proven to be reproducible and very specific to MVM. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with MVM. MVM DNA could be Quantified in CHO cell as well as culture supernatant. When the real-time 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 MVM.

• IMMUNE NETWORK
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• v.5 no.2
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• pp.89-98
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• 2005

Background: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. Methods: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, $gB_{498-505}$ DNA) and recombinant vaccinia virus $(vvgB_{498-505})$ expressing epitope $gB_{498-505}$ (SSIEF ARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with $gB_{498-505}$ epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. Results: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and $vvgB_{498-505}$, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with $vvgB_{498-505}$ as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly $gB_{498-505}$ DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and $vvgB_{498-505}$. Of particular interest CD8+ T cell-mediated immunity was optimally induced when $vvgB_{498-505}$ was used to prime and gB DNA was used as alternative boost. Especially CD7+ T cell responses induced by such protocol was longer lasted than other protocols. Conclusion: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.

• The Plant Pathology Journal
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• v.40 no.1
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• pp.73-82
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• 2024

Gardenia (Gardenia jasminoides) is a popular and economically vital plant known for its ornamental and medicinal properties. Despite its widespread cultivation, there has been no documentation of plant viruses on gardenia yet. In the present study, gardenia leaves exhibiting symptoms of plant viral diseases were sampled and sequenced by both metatranscriptome and small RNA sequencing. As a consequence, bean common mosaic virus (BCMV) was identified in gardenia for the first time and named BCMV-gardenia. The full genome sequence of BCMV-gardenia is 10,054 nucleotides (nt) in length (excluding the poly (A) at the 3' termini), encoding a large polyprotein of 3,222 amino acids. Sequence analysis showed that the N-termini of the polyprotein encoded by BCMV-gardenia is less conserved when compared to other BCMV isolates, whereas the C-termini is the most conserved. Maximum likelihood phylogenetic analysis showed that BCMVgardenia was clustered closely with other BCMV isolates identified outside the leguminous plants. Our results indicated that the majority of BCMV-gardenia virus-derived small interfering RNAs (vsiRNAs) were 21 nt and 22 nt, with 21 nt being more abundant. The first nucleotide at the 5' termini of vsiRNAs derived from BCMV-gardenia preferred U and A. The ratio of vsiRNAs derived from sense (51.1%) and antisense (48.9%) strands is approaching, and the distribution of vsiRNAs along the viral genome is generally even, with some hot spots forming in local regions. Our findings could provide new insights into the diversity, evolution, and host expansion of BCMV and contribute to the prevention and treatment of this virus.

• Korean Journal of Veterinary Service
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• v.26 no.1
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• pp.73-80
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• 2003

In order to obtain information on murine model for foot and mouth disease virus(FMDV) type Asia 1, we studied whether guinea pig was a suitable model for studying FMDV. Apparently healthy 3 months old albino guinea pigs and unweaned 3 days old Swiss albino mice were used for this study. Total of 8 guinea pigs were divided into the infected(n=5) and control(n=3) groups. The incubation period of FMDV in the guinea pigs were roughly 2 days and the viremia persisted for 3 days in the guinea pigs. Mice inoculated with the plasma from control guinea pigs did not show any sign of viremia. The plasma were titrated by virus neutralization test using suckling mice as an indicator host. The mean virus neutralizing antibody titers of infected guinea pig at 3 DPI, 4 DPI and 5 DPI were log$\_$10/2.16, log$\_$10/ 3.39 and log$\_$10/ 3.44, respectively whereas there was no neutralizing antibody titer in control group. The difference between the mortality pattern and mean virus neutralizing antibody titer of infected and that of control group at day 3, 4, 5 were statistically significant(p<0.0l).

• Korean Journal of Veterinary Service
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• v.28 no.1
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• pp.49-69
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• 2005

More than 30 years have elapsed since the first report of the International Committee on Taxonomy of Viruses (ICTV) was published in 1971. Since that publication, the ICTV recognizes about 1,550 virus species, but some 30,000 virus strains and isolates are being tracked by virologists in different fields of biology. The ICTV is the 'international court' of experts that rules on names and relationships of all virus, but only to the level of species. Virus taxonomy is changing rapidly, with changes ranging from the trivial(use of italics for species names) to profound reorganization driven by the explosion of sequence information. The universal system of viral taxonomy now accepts Linnean-like classification at the levels of order, family, subfamily, genus, and species. The suffix '-virales' identifies an order, Families are identified by the suffix '-viridae' subfamilies are identified by the suffix '-virinae', and genera are identified by the suffix '-virus'. The importance of distinguishing subspecies, strains, and isolates in vaccine development, diagnostics, etc. is recognized, but these lower levels are not formally classified by ICTV. This paper mainly introduces taxonomy and classification of animal viruses on the basis of the seventh report of the ICTV edited by Van Regenmortal et al. in 2000.

• Korean Journal of Veterinary Service
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• v.13 no.1
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• pp.54-63
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• 1990

We have studied about the safety, immunity and protective potency in swine and experimental am mais of two inactivated vaccine produced with NYJ-1-87 strain of ADV that was isolated in Korea. Result obtained through the experiments were summarized as follows. 1. The safe potency of ADV antigens inactivated with BEI and formaline to mouse & guinea pig was on the whole good, but protective potency rates of those to challenge with ADV were 60-75％ without the differences to two antigens. 2. Safety, immunity & protective potency of ADV antigens inactivated with BEI and formaline to swine were on the whole excellent, except for a mild increase of rectal temperature in some pigs after challenge with ADV. 3. When virus excretion of the experimental groups after challenge with ADV was examed by swabbing of nasal, all pigs of control gorup excreted virus from 2 days p.c., partially to 10 days p.c.. But in vaccinated groups, only 25-50% of all pigs of each group excreted virus during experimental periods. 4. Titers of antibodies in swine & quinea pig vaccinated with inactivated ADV antigens become increased after the 1 weeth p.i. showing the highest liters on the 4-5 weeths p.i.

• Safe Food
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• v.6 no.1
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• pp.16-21
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• 2011