• Title/Summary/Keyword: Enzyme-linked immunosorbent assay

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Enzyme-linked immunosorbent assay (ELISA) for the detection of RVS (Retrovirus of Salmonid) (ELISA법을 이용한 연어과 어류의 RVS 검출(Retrovirus of Salmonid) 검출)

  • Oh, Myung-Joo;Yoshimizu, Mamoru
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.169-176
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    • 1996
  • An indirect double antibody enzyme-linked immunosorbent assay (ELISA) was developed for rapid detection of a new virus isolated from abnormally swimming salmonid fish, RVS (Retrovirus of salmonid). Results using brain tissue homogenates, and infected cell cultures are described. The sensitivity of the methods is $10^{2.6}$ $TCID_{50}/100{\mu}l$ of the examined cell culture fluid. The specificity was confirmed by the ELISA inhibition test and virological examinations. Viral antigen could be detected in artificially infected fish tissue homogenates. The assay will allow the diagnosis of RVS-infected fish within a day.

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An Enzyme-Linked Immunosorbent Assay for Carp (Cyprinus carpio) Vitellogenin and Assay for Oestrogenic Chemicals

  • Jeong, Jing-Woon;Park, Eon-Jung;Kim, Andre;Park, Jang-Su;Park, Heung-Jai
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2001.05a
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    • pp.238-239
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    • 2001
  • The egg-yolk precursor vitellogenin(VTG) is secreted by the liver of female and male fish, in response to estrogenic compound. Carp(Cryprinus carpio) vitellogenin of one major protein is 160kDa, two minor proteins is 110kDa and 170kDa. We were induced vitellogenin by inject of 17-estradiol and purified in a two step procedure by separating it from plasma protein precipitated by 35% saturated ${(NH_4)}_2SO_4$ and then from the remainder by Mono-Q chromatography. We found major carp(Cyprinus carpio) vitellogenin band at 160kDa. Effect of different concentration of oestrogen on vitellogenin synthesis in carp(Cyprinus carpio) exposed for 4 weeks. Show differential effects on vitellogenin synthesis 7 days after treatment. Plasma vitellogenin was measured 3 times for each by an enzyme linked immunosorbent assay(ELISA). ELISA was developed for the detection of the egg yolk precursor vitellogenin in plasma of carp(Cyprinus carpio). The ELISAS performance was optimized and characterized.

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Competitive Enzyme-Linked Immunosorbent Assay for Glucose-6-Phosphate Dehydrogenase

  • Kim, Moon-Hee
    • BMB Reports
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    • v.30 no.5
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    • pp.326-331
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    • 1997
  • To construct a competitive ELISA standard curve for the detection of glucose-6-phosphate debydrogenase (G6PD), we used highly purified native G6PD (nG6PD) as both immobilized and soluble antigens and anti-G6PD serum raised against nG6PD as antibody. The polystyrene cuvettes coated with nG6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of nG6PD as competitors followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA did not exhibit the typical sigmoidal dose-response curve characteristic of competition immunoassays under the optimal concentrations of antigen and antibody. The soluble nG6PD used as competitor failed to effectively inhibit the binding of antibodies to the immobilized nG6PD. The addition of NADP, a cofactor of G6PD enzyme, to coating buffer used for immobilizing nG6PD to the cuvettes and PBS-Tween-BSA buffer for diluting competitors did not improve the inhibition of antibody binding to immobilized nG6PD by soluble n/G6PD. The addition of BSA to coating buffer did not increase inhibition, either. Surprisingly, when partially active G6PD (paG6PD), obtained by repeated freeze-thawing, was used as competitor, the antibody binding to either immobilized nG6PD or immobilized paG6PD was inhibited 49-58%. We conclude that an effective competitive ELISA system with nG6PD enzyme and anti-G6PD serum for the detection of G6PD may not be established due to the poor inhibition of antibody binding to immobilized nG6PD by soluble nG6PD under the present assay conditions and that the inhibition may be improved by using an inactivated enzyme as competitor regardless of the type of immobilized antigen used. These results imply that the immobilized nG6PD may undergo denaturation upon binding to the polystyrene cuvettes and that our anti-G6PD serum may recognize denatured enzyme better than active enzyme.

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Enzyme-Linked Immunosorbent Assay for the detection of serum lgG and lgM Antibodies to Actinobacillus Actinomycetemcomitans Y4 in Localized Juvenile Periodontitis (Enzyme-linked immunosorbent assay를 이용한 국소적, 유년성 치주염 환자의 혈청내 Actinobacillus actinomycetemcomitans Y4 균주 항체역가에 관한 연구)

  • Jeong, Jong-Pyeong;Jeong, Jin-Hyeong;Choe, Seon-Jin
    • The Journal of the Korean dental association
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    • v.22 no.1 s.176
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    • pp.57-66
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    • 1984
  • Twelve patients of localized juvenile periodontitis were evaluated to detection of serum IgG and IgM antibodies to Actinobacillus actinomycetemcomitans Y4 strain. Sera were isolated from those patients. Antibody titer of patients sera to Aa stran Y4 strain. Sera were isolated from those patients. Antibody titer of patients sera to Aa stran Y4 were determined by modified enzyme-linked immunosorbent assey (ELISA) using sonicated and formalin-fixed whole Aa y4 strain for detection of serum IgG and IgM antibody titers. To compare with health control and L.J.P., we used 12 healthy dental student who did not exhibited any gingivits. Results were determined by using ELISA reader at 400mm absorbance value. Data analysis were performed with comparison of the regression functions relating absorbance to dilution and Dunnett t-test. Significant high antibody titer to As Y4 in L.J.P. sera were shown in this examination(281. 4 Eu-G to 162.80 Eu-G, 106.0 Eu-M to 40.0 Eu-M for sonicated As Y4 antigen and 653.960. to 138.117 Eu-M for intact As Y4) and this data were also statistically significant (P<0.05). This work was supported in part from Seoul national University Hospital Grant and Korean Science Foundation.

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Thermal Inactivation Kinetics of Tyichoderma viride Cellobiohydrolase Determined by Enzyme Linked Immunosorbent Assay and Residual Enzyme Assay (면역학적 방법에 의한 Cellobiohydrolase의 열역학적 특성)

  • 오태광;박관화
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.365-369
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    • 1989
  • Thermal inactivation of Tyichoderma viride cellobiohydrolase was investigated by immunoassay and residual enzyme assay such as carboxymethyl cellulase (CMCase) and filter paper degradation activity (FPase). Arrhenius plots of cellobiohydrolase were appeared as straight line. The Z-values of cellobiohydrolase calculated by CMCase, FPase and immunoassay were 5.2$^{\circ}C$, 6.4$^{\circ}C$ and 5.8$^{\circ}C$, respectively. The thermodynamic parameters obtained from FPase were better agreement with those of immunoassay than CMCase assay.

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