• Title/Summary/Keyword: virus isolation

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Polymerase Chain Reaction for the Detection of Aujeszky's Disease Virus (오제스키병 바이러스 검출을 위한 Polymerase Chain Reaction)

  • Hwang, Dong-hee;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.239-246
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    • 2003
  • Polymerase chain reaction (PCR) was evaluated for the early detection of Aujeszky's disease virus (ADV) DNA from virus-infected cell cultures. For the purposes, the Korean ADV NYJ1-87 was propagated in swine kidney (SK) cells and subjected to the amplification of DNA (217 bp) by PCR using sense and antisense primers specific to gp50 gene of the ADV. In detection of cell-associated viral DNA, reliable PCR conditions were determined as 30 cycles of reaction consisting 1 minute each of denaturation at $94^{\circ}C$, annealing at $55^{\circ}C$ and polymerization at $72^{\circ}C$. The PCR encountered best results with reagent mixtures of $50{\mu}l$ containing $200{\mu}M$ dNTPs, $0.2{\mu}M$ each sense and antisense primers, 1 mM $MgCl_2$ and 10% (v/v) template DNA in the final concentrations. ADV-specific DNAs were detected as early as 6, 6, and 9 hours post-infection, respectively, from lysates of the SK cells infected with ADV of $10^3$, $10^2$ and $10^1\;TCID_{50}/ml$ by this condition. In culture supernatant, the DNAs were detected from ADV of as low infectivity as $10^ {-3}\;TCID_{50}/ml$ by the reduced reagent concentrations and 30 cycles of 1 minute each of denaturation at $94^{\circ}C$ and annealing at $55^{\circ}C$, and 2 minutes of polymerization at $72^{\circ}C$. The lowest amount of detectable ADV DNA was 1 fg. In conclusion, the PCR condition established in the present study was recognized as a feasible alternative to time-consuming procedures in isolation and characterization of the virus.

Genetic and biological characteristics of recent Korean isolates of avian influenza virus subtype H9N2

  • Acharya, Madhav Prasad;Kwon, Hyuk-Joon;Kim, Il-Hwan;Lee, Youn-Jeong;Kim, Jae-Hong
    • Korean Journal of Veterinary Research
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    • v.52 no.4
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    • pp.223-230
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    • 2012
  • The worldwide distribution and continuing genetic mutation of avian influenza virus (AIV) has been posed a great threat to human and animal health. A comparison of 3 isolates of AIV H9N2, A/chicken/Korea/KBNP-0028/00 (H9N2) (KBNP-0028), A/chicken/Korea/SNU8011/08 (H9N2) (SNU 8011) and an inactivated oil vaccine strain A/chicken/Korea/01310/01 (H9N2) (01310), was performed. The former 2 AIVs were isolated from field cases before and after the application of an inactivated H9N2 vaccine in 2007, respectively. The antigenic relationship, viral shedding, tissue tropism and genetic analysis were examined. The comparison of virus shedding from the cloaca and the oropharynx revealed that both isolates were more frequently isolated from the upper respiratory tract (90~100%) 1 day post inoculation (DPI) compared with isolation 5 DPI from gastrointestinal tracts (10~60%). Moreover, the isolate KBNP-0028 were recovered from all organs including bone marrow, brain and kidneys, indicating higher ability for broad tissue dissemination than that of SNU 8011. KBNP-0028 replicated earlier than other strains and with a higher titer than SNU 8011. In full-length nucleotide sequences of the NA gene and a partial sequence of the HA gene of SNU 8011, we found that there might be significant changes in tissue tropism, virus replication and genetic mutation in AIV H9N2 isolates.

Isolation and Biological Characterization of Barley mild mosaic virus(BaMMV) Mild and Severe Strains in Korea

  • Jonson, Gilda;Park, Jong-Chul;Noh, Tae-Hwan;Kim, Mi-Jung;Hyun, Jong-Nae;Kim, Jung-Gon
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.329-333
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    • 2006
  • Two distinct and stable isolates of Barley mild mosaic virus(BaMMV) designated as Naju82-S(severe) and Naju82-M(mild) were obtained. These two isolates differed in their symptomatology, virus transmission characteristics and cultivar specificity at various temperature. Thus, these isolates were referred to as strains in this study. BaMMV Naju-S strain showed severe mosaic symptoms accompanied by necrosis on the infected leaves. Naju82-S strain is more virulent demonstrated by shorter incubation period and relatively high virus concentration than Naju82-M strain. Five Korean cultivars were tested for their pathogenicity to different strains based on the rate of infection. Results showed that infection rate of cultivars to both strains did not significantly differed from each other. However, under different temperatures, the pathogenicity on the two cultivars such as cultivars Hopumbori and Sessalbori were significantly affected. Hopumbori was moderately resistant to both strains at $10-12^{\circ}C$ and susceptible at $15-18^{\circ}C$. Similarly, Sessalbori was moderately resistant at $10-12^{\circ}C$ to both strains but distinctly differentiated at $15-18^{\circ}C$ wherein it was resistant to mild strain and highly susceptible to severe strain. Other cultivars including Baegdong, Jinyangbori and Neahanssalbori consistently showed susceptible reaction to both strains at varying temperatures tested in this study.

Ultrastructural Changes in Enterocyte infected with Porcine Epidemic Diarrhea Virus (돼지 유행성 설사증 (Porcine Epidemic Diarrhea) 감염 소장상피세포의 전자현미경 관찰)

  • Park, Nam-Yong;Koo, Kyoung-bon;Kim, Seung-Jae;Chung, Chi-Young;Park, Young-Seok;Cho, Kyoung-Oh;Kim, Yong-Hwan;Im, Hyung-Ho
    • Korean Journal of Veterinary Pathology
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    • v.3 no.1
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    • pp.35-44
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    • 1999
  • The morphologic changes of small intestinal epithelium in pigs diagnosed as porcine epidemic diarrhea(PED} by virus isolation and immunohistochemistry were studied through light microscope and transmissible electron microscope. On semi-thin section, the histologic findings showed severe villous atrophy and fusion with hyperplasia of cuboidal epithelium in the villi, inflammatory cell infiltration in lamina propria, and increased mitotic figures in the crypt. The structural changes were mostly restricted to the cytoplasm of affected absorptive epithelium of villi. 3 types of epithelial changes were found; degenerated virus-affected cells, undifferentiated cuboidal cells, and normal columnar cells. On electron microscopy, round to spherical viral particles of 50∼l00nm in diameter were found within the dilated vesicles and endoplasmic reticulums of degenerated cells, which had decreased their cytoplasmic electron density due to dilated and missing organelles(e.g. mitochondria, ERs, etc.). Microvilli were shortened and sparse, leaving denuded terminal web of the villous epithelial cells. Fat globules were often found within slightly degenerated enterocytes. On the tip of villi, severely damaged cells were exfoliated and replaced by undifferentiated cuboidal cells We found distinct ultrastructural changes in the jejunal epithelium confirming PED virus infection is involved in malabsorptive diarrhea.

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An Aptamer-Based Electrochemical Sensor That Can Distinguish Influenza Virus Subtype H1 from H5

  • Lee, Jin-Moo;Kim, JunWon;Ryu, Ilhwan;Woo, Hye-Min;Lee, Tae Gyun;Jung, Woong;Yim, Sanggyu;Jeong, Yong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.2037-2043
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    • 2017
  • The surface protein hemagglutinin (HA) mediates the attachment of influenza virus to host cells containing sialic acid and thus facilitates viral infection. Therefore, HA is considered as a good target for the development of diagnostic tools for influenza virus. Previously, we reported the isolation of single-stranded aptamers that can distinguish influenza subtype H1 from H5. In this study, we describe a method for the selective electrical detection of H1 using the isolated aptamer as a molecular probe. After immobilization of the aptamer on Si wafer, enzyme-linked immunosorbent assay (ELISA) and field emission scanning electron microscopy (FE-SEM) showed that the immobilized aptamer bound specifically to the H1 subtype but not to the H5 subtype. Assessment by cyclic voltammetry (CV) also demonstrated that the immobilized aptamer on the indium thin oxide-coated surface was specifically bound to the H1 subtype only, which was consistent with the ELISA and FE-SEM results. Further measurement of CV using various amounts of H1 subtype provided the detection limit of the immobilized aptamer, which showed that a nanomolar scale of target protein was sufficient to produce the signal. These results indicated that the selected aptamer can be an effective probe for distinguishing the subtypes of influenza viruses by monitoring current changes.

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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Isolation of porcine epidemic diarrhea virus (PEDV) in Korea (돼지 유행성 설사 바이러스 (porcine epidemic diarrhea virus)의 국내 분리주 작성에 관한 연구)

  • Kweon, Chang-hee;Kwon, Byung-joon;Jung, Tae-sung;Kee, Young-jin;Hur, Dong-ho;Hwang, Eui-kyung;Rhee, Jae-chin;An, Soo-hwan
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.249-254
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    • 1993
  • The etiological survey on porcine epidemic diarrhea virus(PEDV) by immunofluorescence antibody test(IFA) showed the positive rusult from the intestines of piglet died from acute diarrhea. The viral agent of PED was also isolated from intestine, which showed positive reaction by immunofluorescence test. After passage in Vero cell, the viral agent was further cloned by plaque purification and designated as KPEDV-9. The immunoblotting analysis using hyperimmune sera and porcine sera revealed the presence of several polypetide bands with molecular weight(M.W.) of 88K, 74K, 70K, 58~54, 54~46K, 44~40K and 33~32K, respectively.

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Distribution and Isolation of Soil borne Wheat Mosaic Virus in Korea

  • Lee, Kui-Jae;Lim, Hyun-Suk;Kim, Hyung-Moo;Lee, Wang-Hyu
    • Plant Resources
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    • v.4 no.1
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    • pp.41-47
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    • 2001
  • This study was conducted to investigate the occurrence of Soil borne wheat mosaic virus(SbWMV) in barley fields in Korea and to examine the host pathogenicity of SbWMV. By using the ELISA test, SbWMV was detected in the six regions : Suwon, Milyang, Jinju, Youngkwang, Iksan, and Chonju. SbWMV was isolated from the two strains, Albori strain from Jinju and Eunpamil strain from Milyang. SbWMV was collected from leaves showing mosaic, yellowing and necrosis stripes. SbWMV was inoculated mechanically on 1∼1.5 leaf stages with leaf-rubbing to identify the host pathogenicity of 36 Korean barley cultivars, a wheat cultivar, two rye cultivars, three Japanese barley cultivars and Chenopodium amaranticola. Viral sympoms of inoculated leaves appeared on moulted loaves about 4 to 6 weeks of inoculation. Baegdong and Tapgolbori, infected from Albori strain and Eunpamil strain infected from Samdobori showed much higher susceptibility than C. amaranticola and C. quinoa which showed ring spots and chlorotic spots respectively. Virus particles were observed by the electron microscope. They were rod-shapes, which are bipartite, of 142 nm or 281 nm in length with 20 nm diameter on infected leaves. Specific detection and identification of SbWMV was set up using the RT-PCR. PCR fragments of SbWMV(0.5kb) were obtained by using the designed primers for SbWMV RNA 2.

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Knowledge and Attitudes Not Strongly Related to COVID-19 Prevention, Behaviors, and Vaccination Among Filipino College Students

  • De Castro, Gefilloyd L.;Eijansantos, Abee M.;Jalani, Nuriza P.;Amora, Joni Czarina P.;Lantaya, Gualberto A. Jr.
    • Asian Journal for Public Opinion Research
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    • v.10 no.4
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    • pp.316-339
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    • 2022
  • The effects of the COVID-19 pandemic across various sectors are far-reaching, and the education sector is not an exemption. Approved and recognized vaccines are clinically proven to provide protection and have been considered one of the primary mitigations against the virus. Knowledge, attitude, and related practices (henceforth KAP) toward the virus is widely considered to be an essential cognitive key in public health for health intervention, prevention, and promotion. Previous studies revealed a positive correlation between knowledge and attitude. This study investigated the KAP and their relationship among the 857 college students in one state college in the Philippines using an online survey. Using descriptive statistics, the findings showed that the majority of the respondents have been fully vaccinated against COVID-19. Respondents were shown to be knowledgeable about COVID-19, specifically about its transmission, clinical symptoms, treatment, risk groups, isolation, prevention, and control. Positive attitudes toward individual and governmental mitigation and control of the virus were shown. Their scores on practices revealed that they always exhibit protective and preventive behavioral practices. Pearson correlation revealed a weak significant relationship between knowledge and practice, and a fair significant relationship between attitude and practice. No significant relationship is found between knowledge and attitude. This study hopes to find its place in the literature worthy of consideration as empirical support toward understanding a specific group's perceptive risk and preventive behavior toward the virus, policy making, and health intervention and promotion.

Effects of the modified live vaccines against Bordetella bronchiseptica and canine parainfluenza virus (개 전염성 기관기관지염에 대한 modified live vaccine의 방어효과)

  • Park, Young-Il;Roh, In-Soon;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.31 no.1
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    • pp.57-70
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    • 2008
  • The purpose of this study was to investigate the protective effects of the modified live vaccines against canine Bordetella bronchiseptica (Bb) and canine parainfluenza virus (CPIV) in puppies by nasal administration. A total of 24 puppies were classified as 3 groups consisting of 8, and each one was divided into two subgroups; vaccinated (n=4) and unvaccinated (n=4). Group I, group II and group III were challenged with Bb, CPIV, and Bb+CPIV, respectively. In group I vaccinated puppies (n=4) were experimentally challenged with Bb 2 weeks after vaccination and unvaccinated puppies (n=4) were experimentally challenged with Bb alone. The same methods of the above were applied for group II and group III. We carried out several studies including serum tests, isolation, and histopathological examination. The vaccinated puppies showed higher antibody titers than unvaccinated puppies and the titer sustained during the experimental period. In the isolation test, recovery period was shorter in the vaccinated subgroup than in the other. In clinical signs, the unvaccinated puppies showed the typical signs of tracheobronchitis (coughing, nasal and occular discharge), but another subgroup showed delayed incidence and mild clinical signs. The typical gross lesions and histopathological findings were observed in the unvaccinated puppies. In immunohistochemical findings, the vaccinated puppies showed little intensive in reaction for CPIV antigen than the other. It could be concluded that intranasal vaccination of modified live Bb and CPIV vaccine to puppies is effective to prevent canine infectious tracheobronchitis.