• Title/Summary/Keyword: virus assay

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Detection of bovine rotavirus antigen by enzyme linked immunosorbent assay (효소면역법에 의한 소 로타바이러스 항원 검출)

  • 안재문;유기조;이용희;이종인
    • Korean Journal of Veterinary Service
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    • v.19 no.1
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    • pp.30-38
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    • 1996
  • Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

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Characteristics of Tobacco Mosaic Virus Isolated from Wasabi (Eutrema wasabi) in Korea

  • Kim, Hyung-Moo;Lee, Kui-Jae
    • The Plant Pathology Journal
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    • v.15 no.4
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    • pp.247-250
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    • 1999
  • Wasabies showing mosaic symptoms were collected and extracted for virus purification. Tobacco mosaic virus (TMV) was identified as causal agent by electron microscopy and nucleic acid and coat protein analyses. TMV strains were determined by enzyme-linked immunosorbent assay (ELISA). TMV was identified as W and C strain in wasabi. The results of host reaction indicated that this virus induced local lesions on Nicotiana tabacum cv. Bright Yellow and N. glutinosa, leaf spots on Chenopodium amaranticolor and mosaic symptoms on wasabi. Rot shape virus particles were observed and was about 300 nm in length. About 6.5 kb single RNA molecule was observed from extracted viral RNA sample and 26 KDa coat protein was detected in denatured acrylamide gel. Infection ratio of TMV was 8% for the first cultivation year, but was 22% for the second year when TMV-W antiserum was used. The results of this experiment showed that infection ratios of both TMV-W and TMV-C strains were higher compared to that of TMV-P strain.

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Virus-Cell Fusion Inhibitory Compounds from Ailanthus altissima Swingle (저근백피의 Virus-Cell Fusion 저해활성 성분)

  • Chang, Young-Su;Moon, Young-Hee;Woo, Eun-Rhan
    • Korean Journal of Pharmacognosy
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    • v.34 no.1 s.132
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    • pp.28-32
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    • 2003
  • In order to search for the anti-HIV agents from natural products, eighty MeOH extracts of medicinal plants were applied to a syncytia formation inhibition assay which is based on the interaction between the HIV-1 envelope glycoprotein gp120/gp41 and the cellular membrane protein CD4 of T lymphocytes. Among them, Ailanthus altissima showed a potent virus-cell fusion inhibitory activity. Repeated column chromatoghaphy of the methylene chloride fraction of A. altissima afforded compounds 1$({\beta}-sitosterol-3-O-{\beta}-D-glucoside)$, 2(tetramethoxycoumarin), and 3(ocotillone). Virus-cell fusion inhibitory activity of compound 3(ocotillone) was $70.76{\pm}4.09%$ at the concentration of $100\;{\mu}g/ml$.

Detection of Cymbidium Mosaic Virus and Odontoglosum Ringspot Virus by ELISA and RT-PCR from Cultivated Orchids in Korea (ELISA와 RT-PCR에 의한 국내재배난에서 심비디움 모자이크 바이러스와 오돈토글로섬 윤문 바이러스이 검정)

  • 박원목;심걸보;김수중;류기현
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.130-135
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    • 1998
  • This study was carried out to detect cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV) in cultivated orchid plants in Korea. The standard double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) were carried out for detection of the viruses in the collected orchid samples. ELISA was suitable for massive-scale diagnostic method for virus detection in orchids. RT-PCR was rapid, time-saving and reliable detective method, and detection limit data showed that RT-PCR was 103 times more sensitive than ELISA. Of the 321 individual orchids representing 5 orchids genera tested by the ELISA, CymMV and ORSV were detected in 15.6% and 22.4%, and mixed infection of the both viruses with 4.9%, respectively. Of the Cymbidium plants tested, cultivated plants showed 52.5% virus infection rate with either CymMV or ORSV and both viruses.

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A tendency of herbal medicines on anti-viral effect (항바이러스 효과에 응용되는 한약의 연구 동향)

  • Lim, Sung-Woo
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.199-209
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    • 1998
  • In order to find antiviral effect against Human immunodeficiency virus(HIV), Herpes simplex virus type I(HSV-1) and II(HSV-2) from herb medicines, publicated 29 paters on anti-viral effect of herbal medicines and a convenient virus-induced cytopathic effect (CEP) inhibition assay was introduced. The major virus on experiment are HIV, Hepatitis B virus and HSV-1,2. Those of other studies showed inhibition of infected virus DNA replication and screening test of herbal medicines. More than 15 extractions were prepared by pure water boiling from herbal medicines, and their toxicity of infected cell and anti-viral activities were evaluated. Among them, the major part of herbal medicines showed cell stability compared with the contrast. Cytotoxic concentration (CC) of the $H_2O$ extracts of Padoo against HIV was <4.0, Hyungbangpaedoksan against HIV was 9.3, Whangyonhaedoktang against HIV-1 and HSV-2 was 15.3. These are high level cytotoxic concentration compared with the contrast. But antiviral effect was unable to figure out for selective $index(SI)=CC_{50}/EC_{50}$. The other herbal medicines were unable to showed potent anti-HIV and anti-HSV activity. The antiviral activation using herbs in this thesis have unlimited objects, to select research object will help to show the direction of antiviral drug development that have less side effect and more excellent efficiency.

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Production of monoclonal antibodies against viral hemorrhagic septicemia virus (VHSV, genotype IVa) from olive flounder (넙치 유래 viral hemorrhagic septicemia virus (VHSV, genotype IVa)에 대한 단클론 항체 생산)

  • Jeong, Ha-Na;Jang, Min-Seok;Oh, Myung-Joo;Kim, Wi-Sik
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.149-154
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    • 2017
  • Mouse monoclonal antibodies (MAbs) were produced by using viral hemorrhagic septicemia virus (VHSV, genotype IVa) as an immunogen, isolated from diseased olive flounder (Paralichthys olivaceus). Four hybridoma clones secreting MAbs against VHSV were established. The MAbs were recognized the nucleoprotein (MAb 4), phosphoprotein (MAb 1) and matrix protein (MAbs 2 and 3) of VHSV by western blot analysis. Among them, the MAbs 1 and 4 strongly reacted with the VHSV-infected FHM cells, but not normal FHM cells. In enzyme linked immunosorbent assay, the four MAbs reacted with the VHSV, but not different six fish viruses (infectious hematopoietic necrosis virus, hirame rhabdovirus, spring viraemia of carp virus, infectious pancreatic necrosis virus, marine birnavirus and nervous necrosis virus). These results indicate that the MAbs are useful for diagnosis of VHSV infection.

Antiviral Effect of Amphotericin B on Japanese Encephalitis Virus Replication

  • Kim, Hun;Kim, Seong-Jun;Park, Sue-Nie;Oh, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.121-127
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    • 2004
  • Amphotericin B (AmB), an amphipathic polyene macrolide, is an antifungal drug produced by Streptomyces nodosus. Recently, AmB has been shown to exert antiviral activity against rubella virus and human immunodeficiency virus by different mechanisms. In this study, we evaluated the antiviral effect of AmB against Japanese encephalitis virus (JEV) and investigated which step of the viral life cycle was inhibited by AmB to understand the mechanism of antiviral action of AmB. AmB reduced both plaque size and number in the infected cells in a dose-dependent manner. In addition, a 200-fold reduction of infectious virus titer was observed by treatment of infected cells with $5\mug/ml$ of AmB. AmB acted at the post virus-infection step, but not during adsorption of virus to host cells. Western blot analysis revealed that the accumulated level of JEV envelope protein dramatically decreased in the infected cells by treatment with $5-10\mug/ml$ of AmB. Our results indicate that AmB inhibits the replication of JEV at the postinfection step by interfering with viral replication and/or by inhibiting the synthesis of viral proteins.

Experimental infection of piglets with a field isolate of Aujeszky's disease virus in Korea: Pathogenecity, excretion, distribution and immunogenicity of virus (국내분리 Aujeszky's disease virus의 실험적 감염 자돈에 대한 바이러스학적 연구)

  • Park, Jeong-woo;Jun, Moo-hyung;An, Soo-hwan
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.177-186
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    • 1990
  • To investigate the etiology, pathogenicity and virological properties of NYJ-1-87 strain of Aujeszky's disease virus (ADV) that was isolated from the diseased piglet in Korea, the virus at $10^{6.0}TCID_{50}/0.1ml$ was inoculated intranasally and subcutaneously into 30 to 35 days-old piglets. Results obtained through the experiments were summarized as follows. 1. Ten of the infected piglets were clinically observed for 15 days. On the 2nd day post-inoculation(pi), the signs of pyrexia, anorexia and convulsion were noted. On the 4th to 7th days pi, nervous signs of incoordination and intermittent spasm were shown in the most of piglets, and one out of 5 piglets infected intranasally was died with severe nervous signs at the 7th day pi. The signs became relieved on the 8th day pi and all of remainder were completely recovered on the 13th to 14th days pi. 2. In hematological study, prominent decrease in the number of total leukocyte and lymphocyte was shown in the ADV-infected piglets on the 6th day pi. On the 8th day pi, the cell numbers were slightly increased and returned to normal level on the 10th day pi. 3. Viral excretion of the ADV-inoculated piglets was examined by swabbing of nasal and oral cavities, and rectal feces. During the periods of the 3rd to 11th days pi, the virus was excreted intermittently from nasal and oral cavities, and rectal feces. The nasal excretions were shown the highest virus concentration of $10^{5.2}TCID_{50}/0.1ml$ at the 5th day pi. 4. Recovery of the inoculated virus from various organs of the piglets that were died or experimentally slaughtered was attempted, and the virus was isolated from the tissues of brain and tonsil by the cultured cell-inoculation method. The highest recovery rate was noted in the tonsil. By indirect immunofluorescence antibody assay using ADV-monoclonal antibody, the viral antigens were detected in tissues of spleen and liver as well as brain and tonsil on the 7th to 9th days pi. The virus was not isolated from blood and the tissues of lung and kidney throughout the experiments. 5. Titers of virus neutralizing antibody in the piglets experimentally infected with ADV became increased after the 6th to 9th days pi in both of intranasal and subcutaneous inoculation showing the highest titers of 64 to 128 on the 29th day pi. When the antibody levels were measured by radial immunodiffusion enzyme assay, the reactive diameter was enlarged to be positive after the 4th to 6th days pi in both of intranasal and subcutaneous inoculation showing the largest diameter of 13 to 14mm on the 29th day pi.

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Reverse transcription loop-mediated isothermal amplification assay for the rapid and simultaneous detection of H5 and other subtypes of avian influenza viruses

  • Park, Yu-Ri;Kim, Eun-Mi;Han, Do-Hyun;Kang, Dae-Young;Yeo, Sang-Geon;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.15-20
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    • 2017
  • A two-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was designed for the rapid visual detection of the M gene of all subtypes of avian influenza virus (AIV) and the H5 gene of the H5 subtype of highly pathogenic AIV (HPAIV). The reaction carried out in two tubes in a single step at $58^{\circ}C$ for 40 min, and the assay results could be visually detected by using hydroxynaphthol blue dye. Using M or H5 gene-specific primers, the assay successfully detected all subtypes or H5 subtypes of AIVs, including the Korean representative H5N1 and H5N8 HPAIVs. The detection limit of the assay was approximately $10^{2.0}$ $EID_{50}/reaction$ for the M and H5 genes of H5N1 HPAIV, respectively, and was more sensitive than that of previously reported RT-LAMP and comparable to that of real-time RT-PCR. These results suggest that the present RT-LAMP assay, with its high specificity, sensitivity, and simplicity, will be a useful diagnostic tool for surveillance of currently circulating H5 HPAIVs and other subtypes of AIV in bird population, even in under-equipped laboratories.

The Recency Period for Estimation of Human Immunodeficiency Virus Incidence by the AxSYM Avidity Assay and BED-Capture Enzyme Immunoassay in the Republic of Korea

  • Yu, Hye-Kyung;Heo, Tae-Young;Kim, Na-Young;Wang, Jin-Sook;Lee, Jae-Kyeong;Kim, Sung Soon;Kee, Mee-Kyung
    • Osong Public Health and Research Perspectives
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    • v.5 no.4
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    • pp.187-192
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    • 2014
  • Objectives: Measurement of the incidence of the human immunodeficiency virus (HIV) is very important for epidemiological studies. Here, we determined the recency period with the AxSYM avidity assay and the BED-capture enzyme immunoassay (BED-CEIA) in Korean seroconverters. Methods: Two hundred longitudinal specimens from 81 seroconverters with incident HIV infections that had been collected at the Korea National Institute of Health were subjected to the AxSYM avidity assay (cutoff = 0.8) and BED-CEIA (cutoff = 0.8). The statistical method used to estimate the recency period in recent HIV infections was nonparametric survival analyses. Sensitivity and specificity were calculated for 10-day increments from 120 days to 230 days to determine the recency period. Results: The mean recency period of the avidity assay and BED-CEIA using a survival method was 158 days [95% confidence interval (CI), 135-181 days] and 189 days (95% CI, 170-208 days), respectively. Based on the use of sensitivity and specificity, the mean recency period for the avidity assay and BED-CEIA was 150 days and 200 days, respectively. Conclusion: We determined the recency period to estimate HIV incidence in Korea. These data showed that the nonparametric survival analysis often led to shorter recency periods than analysis of sensitivity and specificity as a new method. These findings suggest that more data from seroconverters and other methodologies are needed to determine the recency period for estimating HIV incidence.