• Title/Summary/Keyword: Virus concentration

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The Comparison of Results Among Hepatitis B Test Reagents Using National Standard Substance (국가 표준물질을 이용한 B형 간염 검사 시약 간의 결과 비교)

  • Lee, Young-Ji;Sim, Seong-Jae;Back, Song-Ran;Seo, Mee-Hye;Yoo, Seon-Hee;Cho, Shee-Man
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.203-207
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    • 2010
  • Purpose: Hepatitis B is infection caused by Hepatitis B virus (HBV). Currently, there are several methods, Kits and equipments for conducting Hepatitis B test. Due to ununiformed methods, it would cause some differences. To manage these differences, it needs process evaluating function of test system and reagent using particular standard substance. The aim of this study is to investigate tendency of RIA method's reagent used in Asan Medical Center through comparing several other test reagents using national standard substance. Materials and Methods: The standard substance in National Institute of Food and Drug Safety Evaluation's biology medicine consists of 5 things, 4 antigens and 1 antibody. We tested reagents using A, B company's Kits according to each test method. All tests are measured repeatedly to obtain accurate results. Results: Test result of "HBs Ag Mixed titer Performance panel" is obtained match rate compared S/CO unit standard with RIA method and EIA 3 reagents, CIA 2 reagents is that company A's reagent is 94.4% (17/18), 83.3% (15/18), B is 88.9% (16/18), 77.8% (14/18). Test result of "HBs Ag Low titer Performance panel" is obtain that EIA 2 reagents is shown 7 posive results, CIA 3 reagents is 11, and RIA method's company A's reagent is 3, B is 2 of 13 in low panel. "HBV surface antigen 86.76 IU/vial" tested dilution. A is obtain positive results to 600 times(0.14 IU/mL), B is 300 times (0.29 IU/mL). Case of "HBV human immunoglobulin 95.45 IU/vial", A is shown positive result to 10,000 times (9.5 mIU/mL) and B is 4,000 times (24 mIU/mL). Test result of "HBs Ag Working Standards 0.02~11.52 IU/mL" is shown that Company A's kit concentration level was 0.38IU/mL, company B was 2.23 IU/mL and higher level of concentration was positive results. Conclusion: When comparing various test reagents and RIA method according to National Standard substances for Hepatitis B test, we recognized that there were no significant trends between reagents. For hepatitis B virus antigen-antibody titers even in parts of the test up to 600 times the antigen, antibodies to 10,000 times the maximum positive results could be obtained. Therefore, we confirmed that results from Asan Medical Center are performed smoothly by reagents and system for hepatitis B virus test.

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Highly Sensitive Detection of Pathogenic Bacteria Using PDMS Micro Chip Containing Glass Bead (유리비드를 포함한 PDMS 마이크로칩을 이용한 고감도 감염성 병원균 측정에 관한 연구)

  • Won, Ji-Yeong;Min, Jun-Hong
    • KSBB Journal
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    • v.24 no.5
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    • pp.432-438
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    • 2009
  • Here, we demonstrated simple nucleic acid, RNA, concentration method using polymer micro chip containing glass bead ($100\;{\mu}m$). Polymer micro chip was fabricated by PDMS ($1.5\;cm\;{\times}\;1.5\;cm$, $100\;{\mu}m$ in the height) including pillar structure ($160\;{\mu}m\;(I)\;{\times}\;80\;{\mu}m\;(w)\;{\times}\;100\;{\mu}m\;(h)$, gap size $50\;{\mu}m$) for blocking micro bead. RNA could be adsorbed on micro glass bead at low pH by hydrogen bonding whereas RNA was released at high pH by electrostatic force between silica surface and RNA. Amount of glass beads and flow rate were optimized in aspects of adsorption and desorption of RNA. Adsorption and desorption rate was measured with real time PCR. This concentrated RNA was applied to amplification micro chip in which NASBA (Nucleic Acid Sequence Based Amplification) was performed. As a result, E.coli O157 : H7 in the concentration of 10 c.f.u./10 mL was successfully detected by these serial processes (concentration and amplification) with polymer micro chips. It implies this simple concentration method using polymer micro chip can be directly applied to ultra sensitive method to measure viable bacteria and virus in clinical samples as well as environmental samples.

Determination of the Optimal Concentration of Fetal Bovine Serum for the Growth of Two Insect Cell and Viruses (두 가지 곤충 세포주에 대한 배양 및 바이러스 증식을 위한 최적 FBS 농도 결정)

  • Lee, Jae-Kyung;Koo, Hyun-Na;Woo, Soo-Dong
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.319-324
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    • 2007
  • To determine the optimal concentration of fetal bovine serum (FBS) on the growth of insect cells and the multiplicity of viruses, the growth of cells (Sf21 and Bm5) and viruses were examined on the various concentrations of FBS. In view of the viability, growth speed, proliferation of cells and the amount of FBS, the most proper concentration for the cell culture were 7% and 5% for Sf21 and Bm5, respectively. The multiplicity of viruses at the various concentrations of FBS was similar in both cell lines at 5 days post-infection (p.i.). However, it differed significantly at 2 and 3 days p.i. The proper concentration of FBS were 10% and 3% for Sf21 at 2 and 3 days p.i., respectively, and 5% for Bm5 at both 2 and 3 days p.i. These results suggested that the optimal concentration of FBS should be determined according to the used cell lines and viruses for their optimum production.

Induction of Shoot Primordium in Culture of Garlic (Allium sativum L.) (마늘 배양에 있어서 신초원기 유도)

  • Choi Joo-Soo;Lee Bok-Kyu;Huh Man-Kyu
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.459-463
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    • 2006
  • Cultivated garlic, Allium sativum L. is economically important for leaves and bulbs, which historically were used in Korea for spices and condiments of Korean food as well as medicine crops. This experiment was carried out to investigate the effect of development and differentiation on culture of A. sativum (cv: white 6) by explant position, hormone composition and sucrose concentration in culture media. Culture method was investigated to induce shoot primordium. Culture efficiency was better with lower tissue of foliage leaf in explant position and on the medium with NAA 0.02 + BAP 1.0 mg/l in hormone composition than any other. Precocious shoot and callus were induced from shoot apex. Shoot was efficiently differentiated on 4,000 mg/l sucrose with increasing concentration of BAP. Shoot primordium was also induced with liquid rotary culture by histological observation. Rhizoid was induced from callus tissue cluster on medium with NAA 0.02 + BAP 2.0 mg/l.

Quantitative Method of Rapid Immunochromatographic Assay Kit for HBsAg-screening using Computer Image Analysis (컴퓨터 상 분석을 이용한 HBsAg-screening용 Rapid Immunochromatographic Assay Kit의 정량적 측정법)

  • 신형순;허태련
    • KSBB Journal
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    • v.15 no.3
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    • pp.243-246
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    • 2000
  • One of recent topics in the case of hepatits B virus(HVB) is the value of hepatitis B surface antigen(HBsAg) concentration as a prognostic maker. We developed uantitative method of rapid immunochromatographic assay(ICA) kit for HBsAg using computer image analysis (CIA) for the purpose of home diagnosis. uantitative ICA using CIA demonstrated integrated optical density(IOD) values proportional to log of reference HBsAg concentrations in the range of 2-200 ng/mL and enzyme-linked immunosorbent assay(ELISA) demonstrated the same in the range of 0.1-100 ng/mL however the test results with sample sear showed the same concentration on both kits. Furthermore repeated tests with the same samples revealed that this quantitative ICA using CIA would be reproducible and coefficient of variation(CV) of the results was 1.38~6.30%.

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Screening of Some Plant Extracts for Inhibitory Effects on HIV-1 and Its Essential Enzymes (Human Immunodeficiency Virus Type I에 대한 수종 식물 추출물의 억제활성 검색)

  • Park, Jong-Cheol;Lee, Jong-Ho;Kim, Gyeong-Eup;Jo, Sung-Kee;Byun, Myung-Woo;Hirotsuku, Miyashiro;Masao, Hattori;Yu, Yeong-Beob
    • Korean Journal of Pharmacognosy
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    • v.29 no.4
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    • pp.338-346
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    • 1998
  • In order to elucidate the relationship between anti-HIV-1 enzyme activity and inhibition of HIV-1 replication by natural sources, extracts from some plants using the foods and oriental medicines were tested for inhibitory effects on the viral replication, reverse transcriptase (RT), protease and ${\alpha}-glucosidase$. In the anti-RT test, water extracts of Ficus carica (leaf), Houttuynia cordata (aerial part) and Ixeris tamagawaensis (aerial part) showed more than 79% inhitibion at a concentration of $100\;{\mu}g/ml$. The protease and ${\alpha}-glucosidase-inhibiting$ samples in the screening were water extract of Syringa dilatata (leaf) and methanol extract of Hibiscus syriacus (leaf and stem), which showed more than 40% inhibition at a concentration of $100\;{\mu}g/ml$. In the primary anti-HIV-1 test, water extracts of Equisetum arvense (aerial part), Hibiscus syriacus (leaf), Ixeris tamagawaensis (aerial part) and Pueraira thunbergiana (leaf) showed the potent inhibition against HIV-1 induced cytopathic effects.

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Pretreatment and Rapid Detection Methods for Wastewater-Based Epidemiology (하수역학 구축을 위한 시료 전처리 기술과 신속검출기술)

  • Lee Jai-Yeop;Lee Bokjin;Jesmin Akter;Ahn Chang Hyuk;Kim Ilho
    • Journal of Korean Society on Water Environment
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    • v.39 no.1
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    • pp.102-110
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    • 2023
  • Wastewater Based Epidemiology (WBE) provides useful information not only on the use of illegal drugs in the community, but also on the presence of hygiene and health products and infectious pathogens in sewage facilities. As a consequence of the SARS-CoV-19 virus epidemic in 2019, monitoring the status of the infection is of utmost importance. SARS-CoV-19 was also detected in sewage, and the number and trend of infections in the community suggest that the application of the WBE system would be useful and appropriate. This study introduces a pre-treatment concentration method including viruses in sewage samples. A total of seven methods which were subdivided into methods for adsorption-extraction, ultra-filtration, PEG precipitation, and ultra-centrifugation, and the results for analyzing the recovery rates were included. Meanwhile, it is necessary to pay attention to rapid detection technologies which analyze infectious pathogens at the site of sewage facilities. These can include ELISA, FTIR, SERS, and biosensor based on the detection principle, and the characteristics, advantages, and disadvantages of each were summarized herein. If rapid detection technologies and accurate quantitative analyses are further developed, the use of sewage mechanics in response to pandemic viruses is expected to expand further.

Antiviral effect of fucoxanthin obtained from Sargassum siliquastrum (Fucales, Phaeophyceae) against severe acute respiratory syndrome coronavirus 2

  • Nalae Kang;Seong-Yeong Heo;Eun-A Kim;Seon-Heui Cha;Bomi Ryu;Soo-Jin Heo
    • ALGAE
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    • v.38 no.4
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    • pp.295-306
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    • 2023
  • Human coronavirus diseases, particularly severe acute respiratory syndrome coronavirus 2, still remain a persistent public health issue, and many recent studies are focusing on the quest for new leads against coronaviruses. To contribute to this growing pool of knowledge and explore the available marine natural products against coronaviruses, this study investigated the antiviral effects of fucoxanthin isolated from Sargassum siliquastrum-a brown alga found on Jeju Island, South Korea. The antiviral effects of fucoxanthin were confirmed in severe acute respiratory syndrome coronavirus 2-infected Vero cells, and its structural characteristics were verified in silico using molecular docking and molecular dynamic simulations and in vitro colorimetric method. Fucoxanthin inhibited the infection in a concentration-dependent manner, without showing cytotoxicity. Molecular docking simulations revealed that fucoxanthin binds to the angiotensinconverting enzyme 2-spike protein (binding energy -318.306 kcal mol-1) and main protease (binding energy -205.118 kcal mol-1). Moreover, molecular dynamic simulations showed that fucoxanthin remains docked to angiotensin-converting enzyme 2-spike protein for 20 ns, whereas it breaks away from main protease after 3 ns. Also, the in silico prediction of the fucoxanthin was verified through the in vitro colorimetric method by inhibiting the binding between angiotensinconverting enzyme 2 and spike protein in a concentration-dependent manner. These results indicate that fucoxanthin exhibits antiviral effects against severe acute respiratory syndrome coronavirus 2 by blocking the entry of the virus. Therefore, fucoxanthin from S. siliquastrum can be a potential candidate for treating coronavirus infection.

General Pharmacology of CJ-50005 (Hepatitis A Vaccine) (A형 간염 예방백신 CJ-50005의 일반약리작용)

  • 김영훈;최재묵;정성학;정용주;이성희;김의경;김제학;박병훈
    • Biomolecules & Therapeutics
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    • v.7 no.4
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    • pp.390-396
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    • 1999
  • CJ-50005 is a hepatitis A vaccine which is prepared by formalin inactivation of the HM175 virus cultured in human diploid MRC-5 cells. The general pharmacological properties of CJ-50005 were evaluated in various animals and in vitor system. CJ-50005 at the doses of 0.025, 0.25 and 0.75 $\mu\textrm{g}$/kg, i.m. had no effect on general behavior in mice, chemo- and electro-convulsions in mice, writhing syndrome induced by acetic acid in mice, the barbital sleeping time in mice, body temperature in rats, charcoal meal propulsion in mice and urine and electrolytes excretion in rats. In anesthetized dogs, CJ-50005(0.25 and 0.75 $\mu\textrm{g}$/kg, i.v) did not alter the respiratory rate, blood pressure, heart rate, femoral blood flow and ECG. In in vitro experiment, CJ-50005 at the concentration up to 0.02 $\mu\textrm{g}$/ml did not produce any changes in the contractions of the isolated ileum of guinea pigs caused by acetylcholine, histamine or $BaCl_2$. Since these pharmacological effects of CJ-50005 were observed at dose much greater than those in clinical use (approximately 0.025 $\mu\textrm{g}$/kg, i.m.), it is likely that this vaccine may be relatively free of undesirable effects in clinical practice.

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Nitro oxide in human cytomegalovirus replication and gene expression

  • Lee, Jee-Yeon;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.152-157
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    • 1997
  • Infection of human fibroblast (HF) cells with human cytomegalovirus (HCMV) result in changes in the intracellular level of second messengers. Since nitric oxide (NO) production has been known to be related with other second messengers, it is probable that HCMV infection of HF cells may involve NO. To test this possibility, the amount of NO was measured following ogenous addition of NO generators such as sodium nitroprusside (SNP) or S-nitroso-N-a-cetylpenicillamine (SNAP) immediately after HCMV infection, however, inhibited virus multiplication. Furthermore, immunoblot experiment using monoclonal antibody to HCMV major immediate early (MIE) proteins or CAT assay using pCMVIE/CAT (plasmid containing CAT gene driven by HCMV MIE promoter) revealed that SNP or SNAP blocked the MIE gene expression. SNP was more effective than SNAP in hibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP in inhibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP. Although the mechanism for the inhibition of HCMV multiplication and MIE gene expression by NO is still elusive some correlation with NO-mediated inhibition of HCMV-induced increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/]) was observed. The increase of [Ca$\^$2+/] following HCMV infection was inhibited by SNP, and less effectively by SNAP. Raising [Ca$\^$2+/ with bromo-A23187 partially reversed the SNP block of MIE gene expression. Thus, there appear to e some relationships among NO. [Ca$\^$2+/], and HCMV MIE gene expression.

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