Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0019163 (hepatitis B)
 38,309 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A new enzyme-linked fluorescence assay (ELFA) suitable for use with peroxidase-antibody conjugates is described. The substrate for the assay is p-hydroxyphenylacetic acid, the fluorescent product of which is stable and unaffected by light. The assay compared favourably with a standard ELISA for the quantitation of IgM antibodies to hepatitis B core antigen.
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PMID:A new enzyme-linked fluorescence assay (ELFA) for use with peroxidase-antibody conjugates: a comparison with ELISA for the quantitation of IgM antibodies to hepatitis B core antigen. 351 41

The envelope of hepatitis B virus is coded for by pre-S1, pre-S2 regions and the S gene. A method was developed to determine antibody to the product of pre-S1 region (anti-pre-S1) and antibody to the product of pre-S2 region (anti-pre-S2), either of IgM or IgG class, by a solid-phase enzyme immunoassay. For the determination of anti-pre-S1, tubular particles containing translation products of pre-S1, pre-S2 regions and the S gene were broken into constituent envelope polypeptides and immobilized on a solid support. Serums were absorbed with spherical particles containing translation products of pre-S2 region and the S gene, obtained from plasma positive for hepatitis B e antigen (HBeAg) and deprived of particles carrying pre-S1 product by an affinity column. They were then tested for the binding with tubular polypeptides fixed on a solid support, and the bound antibody representing anti-pre-S1 was detected by monoclonal antibody to human IgM/mu or IgG/gamma labeled with horseradish peroxidase. For the determination of anti-pre-S2, test serums were absorbed with spherical particles containing the product of the S gene, obtained from plasma positive for antibody to HBeAg and deprived of particles bearing pre-S2 product by an affinity column. They were then tested for the binding with polypeptides, fixed on a solid support, composed of products of pre-S2 region and the S gene. The assay was applied to the determination of anti-pre-S1 and anti-pre-S2 of IgM or IgG class in asymptomatic carriers and in persons who had recovered from infection with hepatitis B virus.
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PMID:A solid-phase enzyme immunoassay for the determination of IgM and IgG antibodies against translation products of pre-S1 and pre-S2 regions of hepatitis B virus. 353 38

We describe an artificial substrate system for optimization of labeling parameters in electron microscope immunocytochemical studies. The system involves use of blocks of glutaraldehyde-polymerized BSA into which a desired antigen is incorporated by a simple soaking procedure. The resulting antigen-impregnated artificial substrate can then be fixed and embedded identically to a piece of tissue. The BSA substrate can also be dried and then sectioned for immunolabeling with or without chemical fixation and without exposing the antigen to dehydrating agents and embedding resins. The effects of various fixation and embedding procedures can thus be evaluated separately. Other parameters affecting immunocytochemical labeling, such as antibody and conjugate concentration, can also be evaluated. We used this system, along with immunogold labeling, to determine quantitatively the optimal fixation and embedding conditions for labeling of hepatitis B surface antigen (HbsAg), human IgG, and horseradish peroxidase. Using unfixed and unembedded HBsAg, we were able to detect antigen concentrations below 20 micrograms/ml. We have shown that it is not possible to label HBsAg within resin-embedded cells using conventional aldehyde fixation protocols and polyclonal antibodies.
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PMID:An artificial test substrate for evaluating electron microscopic immunocytochemical labeling reactions. 359 71

A biotin/avidin solid-phase enzyme immunoassay for the detection and quantitation of the antibody to hepatitis B surface antigen (anti-HBs) is described. The assay utilizes hepatitis B surface antigen (HBsAg) as a solid-phase 'capture' reagent and a mixture of biotinylated HBsAg and avidin-conjugated horseradish peroxidase as a probe 'detector' reagent. The assay was compared to a commercial radioimmune assay for anti-HBs detection. The two assays were found to measure the same molecules and to correlate well regarding anti-HBs titers.
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PMID:A biotin/avidin solid-phase sandwich enzyme immunoassay for the antibody to hepatitis B surface antigen (anti-HBs). 361 Dec 87

A biotin-antibiotin solid-phase enzyme-linked immunoassay for the detection and quantitation of the antibody to hepatitis B surface antigen (anti-HBs) is described. The assay utilizes hepatitis B surface antigen as a solid-phase 'capture' reagent and a mixture of biotinylated HBsAg and antibiotin-conjugated horseradish peroxidase as a detector reagent. The assay was compared to a commercial enzyme immunoassay (AUSAB EIA) which used the biotin-avidin system for anti-HBs detection. The two assays were found to measure the same molecules and to correlate well regarding anti-HBs titers.
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PMID:The utilization of biotin-antibiotin interaction for the detection of antibody to hepatitis B surface antigen (anti-HBs). 361 Dec 88

The compositions of lymphocytes in peripheral blood and liver biopsies from 29 patients with hepatitis B e antigen-positive type B chronic hepatitis were studied by an indirect peroxidase-labeled antibody method using monoclonal antibodies to surface antigens on pan T cells (Leu-1 +), cytotoxic/suppressor T cells (Leu-2a +), helper/inducer T cells (Leu-3a +), natural killer/killer cells (Leu-7 +), and B cells (Leu-10 +). In the peripheral blood during the acute exacerbation of chronic hepatitis B, the percentage of cytotoxic/suppressor T cells was decreased, and the ratio of helper/inducer to cytotoxic/suppressor cells was elevated corresponding to the peak of serum transaminase. In contrast, in liver biopsies obtained during acute exacerbation, numerous lymphocytes infiltrated sites of liver cell necrosis and were predominantly cytotoxic/suppressor cells. When compared with the liver biopsies obtained about 2 months after the peak of serum transaminase, cytotoxic/suppressor cells were significantly increased in those obtained during the acute exacerbation period. No significant change of the percentage of natural killer/killer cells was observed in either the peripheral blood or the liver during the acute exacerbation. These findings suggest that T cell cytotoxicity plays an important role in the mechanisms of liver cell damage in acute exacerbation of chronic hepatitis B.
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PMID:Cellular immune responses in the acute exacerbation of hepatitis B e antigen-positive type B chronic hepatitis. 387 59

In this solid-phase two-site enzyme immunoassay for hepatitis B surface antigen (HBsAg), three monoclonal anti-HBs are used: 5D3 (IgM) is immobilized on plastic beads; 5C3 (IgG2a) and 5C 11 (IgG1), labeled with biotin, are used as the first conjugate. Horseradish peroxidase covalently linked to avidin is the second conjugate. First, serum or plasma is incubated with the antibody-coated bead and biotin-labeled antibodies, simultaneously, at 45 degrees C for 1 h ("stat" procedure), 3 h ("standard" procedure), or 18 h ("overnight" procedure), during which HBsAg forms a complex with the solid-phase antibody and the biotinylated antibodies. The enzyme-conjugated avidin is then bound to the biotin on the antigen-antibody complex at 45 degrees C for 15 min ("stat") or 30 min (standard and overnight procedures). The beads are incubated with enzyme-substrate solution (H2O2 and o-phenylenediamine). Color developed is measured at 492 nm. All procedures satisfied third-generation HBsAg tests required by the FDA Office of Biologics, being sensitive both to ad and ay subtypes in subnanogram amounts. The assay is reactive with adw2, adw4, adr, ayw2, ayw3, and ayr subtypes and can detect viral determinants in HBsAg-anti-HBs immune complex form. Thus it provides a sensitive, simple, and reproducible alternative to radioimmunoassay.
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PMID:A monoclonal-antibody enzyme immunoassay for detection of hepatitis B surface antigen with use of a biotin-avidin system. 388 Dec

Cytoplasmic and cell membrane associated hepatitis B core antigen (HBcAg) were found to be more widespread within infected liver using indirect immunofluorescence on frozen sections than with the widely used direct immunofluorescence method. Fixation of frozen sections with carbon tetrachloride improved tissue histology without reducing the sensitivity of antigen detection. In tissue blocks fixed with formalin or ethanol-acetic acid, detection of HBcAg was reduced in comparison with frozen sections, and many cells containing low concentrations of (usually cytoplasmic and membranous) HBcAg could not be identified even using indirect immunofluorescence or peroxidase-antiperoxidase reactions. In contrast, intracellular hepatitis B surface antigen (HBsAg) was well detected in fixed sections, but membrane associated HBsAg was not detectable after fixation.
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PMID:Widespread presence of cytoplasmic HBcAg in hepatitis B infected liver detected by improved immunochemical methods. 388 5

National Institute of Preventive Medicine (NIPM) has succeeded in the development of an enzyme immunoassay (EIA) kit for detection of hepatitis B surface antigen. The sandwich principle was used for the test. Guinea pig anti-HBs IgG was used for coating microtiter plates and Horseradish peroxidase was conjugated with goat specific anti-HBs. Its stability is longer than 4 months. The lowest detectable dose is 0.7 ng/ml or better for subtype ad of HBsAg tested with Hepatitis Sensitivity Panel, Bureau of Drug and Food, Department of Health. The regression curve was determined by testing 66 samples with Auszyme II (EIA Kit. Abbott Lab.) and NIPM Kit, while Auszyme II used as a reference kit. The two EIA kits correlated well with a coefficient of determination (r2) of 0.86. Evaluation on 1,157 patients' and officers' serum samples in Tri-Service General Hospital showed that the positive rate was 24.7% (286/1,157) by RIA (Clinical Assays, Travenol Lab., USA) and that of NIPM EIA Kit was 24.4% (282/1,157). There was no statistical significance in terms of positive rate (p greater than 0.05). The positive rates of 534 blood donors are 22.1% (118/534) and 21.9% (117/534) respectively. Another evaluation on 974 serum samples in National Taiwan University Hospital showed that the positive rate was 27.2% (265/974) by Ausria II-125 (RIA. Abbott Lab.) and that of NIPM EIA Kit was 27.4% (267/974). The undetectable rate and false positive rate of NIPM EIA Kit were 0.41% (4/974) and 0.62% (6/974) respectively. In comparison with four other kind of commercial EIA Kits, the results of NIPM EIA Kit were satisfactory also. We have scaled up the reagent preparations for the kit, except the antibody coated microtiter plate preparation. At the end of March 1985 we will supply 850 EIA kits for the Development Center for Biotechnology for their hepatitis B vaccine production program.
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PMID:[Clinical evaluation of a domestically prepared enzyme immunoassay kit for the detection of hepatitis B surface antigen]. 389 43

Hepatitis B virus (HBV) core antigen (HBcAg) detected by the peroxidase-antiperoxidase technique was present in the nucleus and cytoplasm of some infected hepatocytes but only in the cytoplasm of other hepatocytes. When cells expressing HBcAg were examined by in situ hybridization for the presence of HBV DNA, the intracellular level of cytoplasmic HBV replicative intermediate DNA correlated with the level of cytoplasmic HBcAg, but not with the presence or absence of nuclear HBcAg. This suggests that nuclear HBcAg may not be directly involved in hepadnavirus replication.
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PMID:Cytoplasmic (but not nuclear) hepatitis B virus (HBV) core antigen reflects HBV DNA synthesis at the level of the infected hepatocyte. 390 90


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