Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The use of alkaline phosphatase immunocytochemical staining was explored for the rapid diagnosis of poliovirus, adenovirus, herpes simplex virus and cytomegalovirus infections in cell cultures. In this test, viral antigens treated with their relative antibody were incubated with alkaline phosphatase-labelled antisera. The enzyme label was developed with a naphthol salt in the presence of a diazonium salt (Fast Blue) in order to obtain a blue coloured precipitate at the site of the enzyme. It is suggested that this immunocytochemical technique is valuable in the detection of viral infections and would be an appropriate test to use when rapid diagnosis is required.
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PMID:Rapid diagnosis of viral infections by an alkaline phosphatase immunocytochemical method. 300 28

The relative sensitivity of two enzyme detection procedures was investigated in a simultaneous "monoclonal" ELISA for herpes simplex virus (HSV). A cyclical enzyme amplified detection system with alkaline phosphatase, rather than horse-radish peroxidase and a conventional chromogenic substrate, gave an increase in absolute sensitivity and a 20 to 30% increase in the detection of HSV in routine isolation-positive genital specimens collected in transport medium. The HSV detection rate, with both procedures, was shown to vary with the site and clinical stage of lesion sampled; it was highest with penile vesicular lesions. Direct extraction of the swab specimen in a small volume of diluent further increased the sensitivity of antigen detection giving positive and negative predictive values of 100 and 96% respectively. The overall sensitivity of HSV detection was equivalent to that obtained by isolation in cell culture. The amplified ELISA offers an alternative, rapid, simple, non-culture technique for routine HSV diagnosis that does not rely upon retention of virus viability.
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PMID:Herpes simplex virus detection by ELISA: effect of enzyme amplification, nature of lesion sampled and specimen treatment. 302 Jan 69

A rapid (3.5 h) enzyme-linked immunosorbent assay (ELISA) was developed for the detection of serum antibodies to herpes simplex virus type 1 (HSV-1) and to two antigenically related monkey viruses, simian agent 8 (SA8) and Herpesvirus simiae (B virus). Crude preparations of detergent solubilized infected cells and similarly treated control mock-infected cells served as antigens for coating wells in microplates. Biotinylated protein A and avidin-conjugated alkaline phosphatase were used to detect antibodies in sera from different species (humans, monkeys and rabbits). Three prototype assays are described with three degrees of specificity. Common or specific determinants on the viral antigens could be assayed in simple competition tests using similar antigen preparations to those coating the wells. The specific assays permitted rapid differential serodiagnosis of antibodies to human and simian herpesviruses.
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PMID:ELISA for detection of group-common and virus-specific antibodies in human and simian sera induced by herpes simplex and related simian viruses. 302 5

A sensitive enzyme-linked immunosorbent capture assay using biotin and streptavidin (capture B/SA ELISA) was developed using type-specific monoclonal antibodies for typing of herpes simplex virus. Rabbit anti-herpes simplex virus immunoglobulin G was used as the capturing antibody, and biotin-linked type-1-specific mouse monoclonal antibody or rabbit type-1- or type-2-specific polyclonal antibody served as the detecting antibody. The captured antigen was detected by an ELISA with alkaline phosphatase-conjugated streptavidin, which reacted with biotin molecules on the detector antibody. The capture B/SA ELISA was compared with other methods for efficiency and reliability in typing. Results obtained by restriction endonuclease digestion of the radiolabeled viral genome were used to determine the type (1 or 2) of clinical isolates. These results were then used as a reference for determining the accuracy of the capture B/SA ELISA, as well as that of the immunofluorescence method, both of which are easily adaptable for use in the clinical laboratory. The three methods were in perfect agreement. It was determined that both the capture B/SA ELISA and the immunofluorescence method using monoclonal antibodies provided typing results with 100% specificity and 100% sensitivity and thus were accurate and reliable. However, the ELISA was the method of choice because of its simplicity, rapidity, and use of nonradioisotopic reagents.
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PMID:Typing of herpes simplex virus by capture biotin-streptavidin enzyme-linked immunosorbent assay and comparison with restriction endonuclease analysis and immunofluorescence method using monoclonal antibodies. 302 48

Short synthetic oligonucleotides have been covalently cross-linked to alkaline phosphatase using the homobifunctional reagent disuccinimidyl suberate. The oligomers, twenty-one to twenty-six bases in length, are complementary to unique sequences found in herpes simplex virus, hepatitis B virus, Campylobacter jejuni and enterotoxigenic Escherichia coli. Each oligomer contains a single modified base with a 12-atom "linker arm" terminating in a reactive primary amine. Cross-linking through this amine results in oligomer-enzyme conjugates composed of one oligomer per enzyme molecule that have full alkaline phosphatase activity and can hybridize to target DNA fixed to nitrocellulose within 15 minutes. The hybrids are detected directly with a dye precipitation assay at a sensitivity of 10(6) molecules (2 X 10(-18) mol) of target DNA in 4 hours development time. The enzyme has no apparent effect on selectivity or kinetics of oligonucleotide hybridization and the conjugates can be hybridized and melted off in a conventional manner.
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PMID:Preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use as hybridization probes. 374 5

A sensitive enzyme-linked immunosorbent capture assay with biotin and streptavidin (capture B/SA ELISA) was developed to detect herpes simplex virus (HSV) antigen. Rabbit anti-HSV antibody (immunoglobulin G fraction) was coated on flat-bottom, irradiated, 96-well polystyrene microtiter plates and served to capture HSV antigen. Clinical specimens from patients with genital herpes were added. Biotin-linked rabbit anti-HSV immunoglobulin G was used as the second antibody. The antigen-antibody complex was detected with alkaline phosphatase-conjugated streptavidin, which linked to the biotin. With clinical specimens, the test had a sensitivity of 95.6% and a specificity of 91.4% when compared with the tissue culture method. The presence of HSV antigen in specimens devoid of infectivity was confirmed by blocking the reaction with unlabeled rabbit and human antibody to HSV. The level of antigen detected by the capture B/SA ELISA did not necessarily correlate with the infectivity titer of the specimens. HSV antigens could be detected by the capture B/SA ELISA when the virus infectivity was destroyed at 37 degrees C, by UV irradiation, or by Triton X-100 treatment, but not when hypochlorite treatment was used. Greater sensitivity was obtained when HSV-1- and HSV-2-specific antibody reagents were used simultaneously in each test. The capture B/SA ELISA provides a relatively rapid method (4.5 h) which is quite sensitive and specific when compared with other non-tissue culture, direct assay methods.
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PMID:Rapid detection of herpes simplex virus in clinical specimens by use of a capture biotin-streptavidin enzyme-linked immunosorbent assay. 608 5

Treatment of VERO cells with 2,4,6-triamino-pyridimyl-5-azobenzene (10 gamma/ml) one hour after inoculation of herpes simplex virus type 1 limits the influence of the virus infection, consisting in the enchancement of the activities of alkaline phosphatase, acid phosphatase and LDH (especially of the latter two enzymes) and in changes in the isoenzyme patterns of acid and alkaline phosphatases and of peroxidase. In general the influence of the drug is expressed both by the fact that enzyme levels in homogenates of infected cells become closer to the values recorded in controls, and by modifications in the isoenzyme patterns altered by virus multiplication, making them more similar to those of uninfected, untreated cells.
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PMID:Influence of 2,4,6-triamino-pyrimidyl-5-azobenzene on the activities of acid phosphatase, alkaline phosphatase and lactate dehydrogenase in cell cultures infected with herpes simplex virus type 1. 625 Feb 80

For setting up an Enzyme-Linked Immunosorbent Assay (ELISA) to detect antibodies to herpes simplex virus type 1 (HSV-1), the virus was propagated in Vero cells and partially purified by sonification and ultracentrifugation on 30% sucrose solution. DEAE ion-exchange column chromatography was used for purification of goat anti-human IgG serum. The anti-human IgG immune serum and alkaline phosphatase were conjugated by glutaraldehyde method. ELISA test was performed by reacting HSV-1 antigen coated in polystyrene tubes with serum specimens and enzyme-IgG conjugates. The color produced by enzyme-substrate reaction was measured on a spectrophotometer. The results obtained by the ELISA had a good agreement with those obtained by a standard neutralization procedure on 119 serum specimens tested for antibody to HSV-1.
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PMID:Detection of serum antibody to herpes simplex virus type I by enzyme-linked immunosorbent assay (ELISA). 626 77

E-5-(2-Bromovinyl)-2'-deoxyuridine (BrvdUrd) and E-5-(2-iodovinyl)-2'-deoxyuridine (IvdUrd) are among the most potent and selective inhibitors of herpes simplex virus type 1 (HSV-1) replication. To elucidate the site of inhibition, we examined whether the halovinyl analogs are incorporated into DNA using two approaches. (i) In assays with purified DNA polymerases omitting dTTP from the reaction system, addition of either BrvdUTP or IvdUTP increased the polymerization reaction, indicating that these two analog triphosphates can be alternate substrates. (ii) When HSV-1-infected Vero cells were grown in the presence of either BrvdUrd or IvdUrd, there was an increase in the density of both the viral and cellular DNA. The viral DNA had 40% of its thymidine moiety substituted by IvdUrd when the concentration of [125I]IvdUrd was 24 microM (in the absence of added thymidine). At 30 microM BrvdUrd and 1 microM [2-14C]thymidine, the viral DNA had only 11% of its thymidine moiety substituted by BrvdUrd, presumably because of the presence of added thymidine. Following digestion of [125I]IvdUrd-substituted DNA with DNase 1, venom phosphodiesterase, and alkaline phosphatase, the radioactivity co-migrated with nonradioactive IvdUrd in thin layer chromatography. Under similar conditions, no detectable incorporation of either [125I]IvdUrd or BrvdUrd into mock-infected Vero cell DNA was observed. Thus, IvdUrd and BrvdUrd are incorporated into DNA of HSV-1 infected cells but not into DNA of uninfected cells.
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PMID:Incorporation of E-5-(2-halovinyl)-2'-deoxyuridines into deoxyribonucleic acids of herpes simplex virus type 1-infected cells. 627 55

Plasma membrane extracts from Herpes simplex virus type 1 transformed hamster embryo fibroblasts were chromatographed on Lens culinaris lectin coupled to Sepharose (LcH-Sepharose) and analysed by dodecyl sulphate polyacrylamide gel electrophoresis. Coomassie blue-staining revealed two major protein bands with apparent molecular weights of 125 000 and of about 75 000-90 000. In plasma membranes isolated from these tumor cells prior labeled with [3H]fucose or [3H]glucosamine these bands contained the highest amounts of incorporated radioactivity. Separation by LcH-Sepharose-affinity chromatography as well as metabolic labeling clearly demonstrates their glycoprotein character. The 125 000 protein coincides with alkaline phosphodiesterase I activity with a Km of 6 . 10(-4) M for TMP p-nitrophenyl ester and is competitively inhibited by UDP-N-acetylglucosamine. This enzymatic activity is also present in normal hamster embryo fibroblasts. Gel electrophoresis of the Lens culinaris lectin-binding glycoproteins from plasma membranes of normal hamster embryo fibroblasts additionally revealed a strong alkaline phosphatase activity represented by an apparent molecular weight of 150 000, while HSV1 hamster tumor cells contain only a very weak activity of this enzyme activity. HSV-lytically infected cells, however, have unchanged levels of alkaline phosphatase activity, whereas alkaline phosphodiesterase activity increases slightly.
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PMID:Alkaline phosphodiesterase I and alkaline phosphatase I in plasma membranes of herpes simplex virus type 1 transformed hamster cells. 627 77


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