Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.11.1.7 (peroxidase)
 65,474 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

For double tracing experiments, wheat germ agglutinin (WGA) molecules labeled with two different haptens are desirable. In the present report the suitability of digoxigenylated WGA (DIG-WGA) for retrograde tracing was investigated. For this purpose the new tracer was pressure injected into rat brains and the transported DIG-WGA visualized via its digoxigenyl group with an alkaline phosphatase linked anti DIG antibody in permanently stained sections of high quality. With fixatives containing 2.5% glutaraldehyde only few positive cells were found. However, at milder fixation conditions (4% paraformaldehyde, 0.05% glutaraldehyde 0.2% picric acid, 30 min) retrogradely labeled cells were detected with a sensitivity comparable to tetramethylbenzidine protocols for conventional WGA-HRP (horseradish peroxidase) tracing. Preliminary experiments suggest excellent suitability for double labeling.
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PMID:Digoxigenylated wheat germ agglutinin visualized with alkaline phosphatase-labeled anti-digoxigenin antibodies--a new, sensitive technique with the potential for single and double tracing of neuronal connections. 170 75

The Diffusion-In-Gel Enzyme Linked Immunosorbent Assay (DIG-ELISA) is a new and simple method for the quantitation of antibodies to diffuse from wells in gel in petri dishes and absorb to an antigen coated to the plastic surface prior to testing. The antigen-antibody reaction is visualized with horseradish-peroxidase conjugated class-specific anti-immunoglobulins. The DIG-ELISA permits detection of 0.1 microns/ml of specific antibodies. This method was used to determine the IgG, IgA and IgM antibody levels to Yersinia enterocolitica O:3 in sera from patients with antibody titres to this microorganism, as determined by direct agglutination and complement-fixation test. The DIG-ELISA IgG antibody values, in contrast to IgA and IgM, correlated well to titres obtained by the direct agglutination method. Low degrees of correlations were found for all three immunoglobulin classes when compared to the complement fixation test.
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PMID:Diffusion-in-gel enzyme linked immunosorbent assay (DIG-ELISA) for detection of antibodies to Yersinia enterocolitica O:3. 678 10

The diffusion-in-gel enzyme-linked immunosorbent assay (DIG-ELISA) is a new and simple method for quantitation of antibodies, based on the ability of antibodies to diffuse from wells in gel and adsorb to antigen which is bound to a polystyrene surface. The antigen-antibody reaction is visualized with a color reaction caused by horseradish peroxidase-conjugated class-specific anti-immunoglobins. This method was used to study the immunoglobulin G, A, and M immune response to Salmonella typhi O antigen in individuals immunized with a monovalent heat-inactivated typhoid vaccine. The antibody values obtained by the DIG-ELISA method correlated with those evaluated by conventional direct agglutination (Widal) and indirect hemagglutination methods. The DIG-ELISA method was also found to be sensitive, specific, and economical, as well as suitable for handling large numbers of sera while requiring very simple equipment.
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PMID:Comparison of diffusion-in-gel enzyme-linked immunosorbent assay with conventional serological methods for detection of class-specific antibodies to Salmonella typhi O antigen. 679 14

To simplify PCR-SSO HLA-DRB generic typing, we labeled eight of 15 oligonucleotide probes with DIG and the others with biotin, and hybridized each dot blot with both a biotin-labeled probe and a DIG-labeled probe simultaneously. We chose oligonucleotide pairs which require the same hybridization and stringent washing conditions and do not compete with each other during hybridization. After incubation with a mixture of anti-DIG Fab fragment-alkaline phosphatase and streptavidin-peroxidase conjugates, specific binding of the DIG-labeled probe was revealed by a chemiluminescent substrate (CSPD) and specific binding of the biotin-labeled probe was subsequently visualized by a chromogenic substrate (TMB). The sensitivity of both probes was similar and gave comparable hybridization signals. Using this simplified procedure, the number of hybridizations or dot blots can be reduced to half the usual amount and the labor involved in PCR-SSO typing significantly reduced.
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PMID:A polymerase chain reaction-sequence-specific oligonucleotide procedure for HLA class II typing using biotin- and digoxigenin-labeled probes simultaneously in hybridization. 818 60

We describe a new, non-radioactive microtitre plate assay for the analysis of genetic variations at the DNA level. The new method combines hybridization of oligonucleotides with PCR amplified DNA in liquid phase with detection in solid phase using an ELISA-reader. Genomic DNA is labelled with digoxigenin during PCR using a nucleotide mix containing DIG-11-2'-deoxy-uridine-5'-triphosphate (DIG-11-dUTP). The DIG labelled, amplified genomic DNA is hybridized in solution with an oligonucleotide which is labelled with one biotin at its 3'-end, using biotin-16,2',3'-dideoxy-uridine-5'-triphosphate (BIO-16-ddUTP) and DNA deoxynucleotidylexo-transferase (TdT). The hybridized complex is immobilized in a streptavidin (SA) coated microtitre plate via the biotin and detection of digoxigenin is performed using anti-digoxigenin horseradish peroxidase, fab fragments (anti-DIG-POD), and the colorimetric substrate 2,2'-Azino-di-(3-ethylbenzthiazolinsulfonat[6]) (ABTS). The resulting absorbtion of the assay is analysed in a microtitre plate reader. This method results in highly specific and sensitive hybridization signals and with the 15 oligonucleotides chosen, allows the typing of DR1-DR10.
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PMID:HLA class II typing in a microtitre plate format using digoxigenin-labelled amplified DNA and biotin-labelled oligonucleotide probes. 909 10

A rapid detection method for the six established genotypes of rabies and rabies-related viral RNA using RT-PCR-ELISA is described. The detection of digoxigenin-labelled amplified products is performed by solution hybridization to two specific, biotin-labelled, capture probes, which are complementary to the inner region of the amplification products. The capture probe and amplified product hybrid are then immobilised on a streptavidin-coated microtitre plate, bound products are detected by an anti-DIG Fab fragment conjugated to peroxidase, and colorimetric reaction automatically measured. This method was up to 100-fold more sensitive than Southern blot hybridization, detecting 0.00002 TCID50/ml of a genotype 1, classical rabies virus strain. The complete detection methodology from RT-PCR to PCR-ELISA detection could be completed within 10 h. Using this procedure, we were 100% successful in detecting 60 isolates from a representative selection of the six established genotypes from all over the world. This test is a useful additional tool for the detection of the rabies and rabies-related viruses, which is easy to perform, rapid and highly sensitive.
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PMID:Rapid detection of rabies and rabies-related viruses by RT-PCR and enzyme-linked immunosorbent assay. 950 52

To detect Mycobacterium tuberculosis in clinical samples, we used the M. tuberculosis-complex specific insertion sequence IS990 as the target in a simple DIG-PCR ELISA assay, as this element is present as a single copy in all strains of M. tuberculosis we have examined to date. The IS990 test was compared with a similar PCR that utilizes IS6110 as target. For detection of PCR product, digoxigenin-11-dUTP (DIG-dUTP) was incorporated into the product. After amplification, the PCR product was hybridized with biotinylated capture probe, which was complementary to the inner part of the amplicon. The hybrid was captured onto streptavidin-coated microtiter plate and DIG-labeled PCR product was detected using a peroxidase-conjugated antibody to DIG. We evaluated DIG-PCR ELISA for the detection of M. tuberculosis DNA in 265 respiratory and non-respiratory specimens taken from patients with known and suspected tuberculosis disease or from controls. The sensitivity and specificity of both IS990-based test and IS6110-based test was 96.5% and 95.3% respectively, comparable to the sensitivity and specificity of the IS6110-based test. The results demonstrate that the IS990 PCR ELISA test is a rapid and sensitive tool for the detection and identification of M. tuberculosis in clinical samples, and may have advantages to the more widely used IS6110-based tests, particularly in areas where IS6110-negative strains are found.
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PMID:Detection of Mycobacterium tuberculosis in clinical samples using insertion sequences IS6110 and IS990. 1158 95

A sensitive non-radioactive microplate hybridization assay for the detection of proviral DNA of bovine leukemia virus (BLV)-specific polymerase chain reaction (PCR) product is described. The PCR products are labeled by adding digoxigenin-dUTP to the nested PCR reaction and are captured by a microtitre plate coated with oligonucleotide probe, which is complementary to the inner region of the amplification product. Captured products are reacted with an anti-DIG Fab fragment conjugated to peroxidase, and detected using a colorimetric reaction. The PCR-enzyme linked immunosorbent assay (ELISA), detecting as low as 10(-4) ng of proviral DNA in a background of 1 microg of BLV-negative DNA, was up to 100-fold more sensitive than ethidium bromide staining, and showed equal sensitivity to Southern blot hybridization. Using this method it was possible to monitor the presence of proviral DNA in four sheep infected experimentally with BLV, over a 10 months postinfection period, as well as in 29 cattle infected naturally. The test is rapid and highly sensitive and is a useful additional tool for the detection of BLV-infected animals.
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PMID:The detection of bovine leukemia virus proviral DNA by PCR-ELISA. 1168 1

An RT-PCR/ELISA system has been developed that detects and differentiates Rinderpest virus (RPV) from the other closely related morbillivirus of ruminants, Peste des petits Ruminants virus (PPRV). In addition, using lineage specific probes, it is possible to determine whether the virus sample is wild-type or vaccine, and the likely origin of the outbreak if it is wild-type. It involves carrying out a RT-PCR with one digoxygenin (Dig)-labelled primer followed by a hybridisation step with a virus-specific, biotin-labelled, probe. The hybridisation step is carried out in an ELISA format on a streptavidin-coated plate. The DIG-labelled products are detected using a specific anti-DIG monoclonal antibody and an anti-mouse horseradish peroxidase conjugate. The hybridisation step replaces nucleotide sequencing or nested PCR for confirmation of the identity of DNA product. The assay is fast and easy to carry out and can give semi-quantitative estimates of the virus content of samples.
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PMID:Rinderpest virus lineage differentiation using RT-PCR and SNAP-ELISA. 1244 35

We established a sensitive non-radioactive in situ hybridization (ISH) method for the detection of chicken IgG gamma-chain mRNA in paraffin sections. RNA probes were transcribed in vitro from cloned chicken IgG CH1 nucleotide sequences with SP6/T7 RNA polymerases in the presence of DIG-UTP. These probes were used for hybridization and were immunodetected using anti-DIG antibodies conjugated to horseradish peroxidase. The immunoreactive products were visualized with DAB-H(2)O(2). IgG gamma-chain mRNA-expressing cells were localized in both the spleen and oviductal tissues. This method demonstrated an excellent sensitivity since the ISH signal was clear and the background was negligible. We found that in the spleen IgG gamma-chain mRNA-expressing cells were present mainly in the red pulp, whereas in the oviduct they appeared mainly in the mucosal stroma and not in the mucosal epithelium.
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PMID:A sensitive non-radioactive in situ hybridization method for the detection of chicken IgG gamma-chain mRNA: a technique suitable for detecting of variety of mRNAs in tissue sections. 1273 84


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