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)

Detection of toxoplasma IgM antibodies by employing enzyme linked immunosorbent assay (ELISA) technique was developed using different enzymes viz, horseradish peroxidase (HRP) (EC. 1.11.17), urease (EC. 3.5.15) and penicillinase (EC 3.5.2.6) as markers. Of these enzymes, HRP is light sensitive and needs dark chamber, also inactivated by preservative sodium azide. Similarly urease test system is extremely pH sensitive and demands special care during ELISA technique. Whereas penicillinase showed certain distinct advantages viz. stable at room temperature, high specific activity and economical. In the present studies it was observed that the sensitivity of penicillinase is similar to HRP and urease, marker enzymes used in commercially available diagnostic kit. The prominent feature of detection of toxoplasma IgM antibodies involving these three enzymes are: a) Shorter incubation time (About 2.5 hours) b) No false positive reaction. Moreover, these enzyme conjugates were prepared from F (ab')2 fragments of antitoxoplasma rabbit serum to elicit specific interaction with IgM antibodies only, avoiding cross interaction with other non-specific proteins like compliment systems and rheumatoid factor.
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PMID:Detection of toxoplasma IgM antibodies by ELISA method: a comparative study of different enzymes as markers. 142 91

This paper describes an original dot-enzyme-linked immunosorbent assay (ELISA) for predicting ovulation in women, based on the detection of the pre-ovulatory estrogen peak in urine. A monoclonal anti-estrogen antibody is used which recognizes not only free estrogens but also some of their urinary metabolites (17-glucuro- and sulfo-conjugates) allowing a direct assay on early morning urines. Antigen is immobilized as a spot on a nitrocellulose membrane which is immersed in urine in the presence of this antibody. A peroxidase-labeled second antibody allows the detection of the first antibody bound to the membrane. Antigen and anti-estrogen antibody concentrations are chosen to obtain a maximal enzymatic coloration of spots corresponding to basal urinary estrogen levels and no coloration corresponding to the pre-ovulatory surge. Six menstrual cycles were studied, comparing dot-ELISA results with patterns of: (1) urinary estrogens measured by RIA either directly or after hydrolysis and extraction, and (2) basal body temperatures. The validity of the pre-ovulatory signal obtained and the requirements for an adaptation of this methodology to a reliable home kit are discussed.
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PMID:A direct dot-enzyme immunoassay to detect human ovulation. 156 85

A number of studies have been published to evaluate the inmunocytochemical assay for ER using monoclonal antibodies. Histoscores so far used consider two variables: the number of cells and the intensity of the reaction. There are however indications that only the proportion of stained cells are important for assessment and show a direct correlation with quantitative data. We studied 77 breast invasive adenocarcinomas stained with the Abbott ERICA kit and used a simple scale of 0 to 4 for the estimation of ER. Tissues were snap frozen in liquid N and immunostained with the specific antibody and peroxidase. Immunostaining was estimated in a simple observational scale from 0-4+ where 0 = no staining or few scattered positive cells; 1+ up to 25%; 2+ up to 50%; 3+ up to 75% and 4+ more than 75% of stained malignant cells. Counts were performed in at least 100 malignant cells in various microscopic fields. Staining was always nuclear and a considerable heterogeneity in the number of cells and the intensity of the reaction was observed. Grading specimens from 0-4+ was found simple and reproducible. In 45% there was no immunostaining and 54% were positive for ER. In patients greater than 50 years of age 67% were positive; in patients less than 50 years of age only 33% had ER. This procedure has many advantages for clinical use: it is simple, it does not require sophisticated equipment, it is reproducible and can be performed in small tissue fragments, such as needle aspiration material, as well as in cytological smears.
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PMID:Evaluation of a simple semiquantitative assay for estrogen receptors in breast cancer. 165 Sep 56

The authors describe a method for the detection of carbohydrate determinants contained by the cellular surface membrane glycosylated biopolymers, based on the use of horse radish peroxidase-conjugated Arachis, Helix pomatia, Ricinus communis, Lens, Glycine max, Sophora japonica lectins. The pattern of lectin receptor distribution on normal human blood lymphocytes, tonsillar lymphoid cells, and in some forms of leukemia and lymphoma is shown. The authors consider it essential that a lectin kit be used along with enzymochemical and immunocytochemical markers as an additional test for the diagnosis of various forms and types of malignant lymphoproliferative diseases.
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PMID:[Determination of lectin receptors on the surface membranes of lymphoid cells]. 169 58

A chemiluminescent method has been offered to assay the blood serum total cholesterol. The method is based on coupled enzymatic reactions in two steps: (1) esterified cholesterol hydrolysis by cholesterol esterase in the presence of sodium cholate and (2) cholesterol oxidation by cholesterol oxidase and chemiluminescent assay of the released hydrogen peroxide in peroxidase reaction with luminol and p-iodophenol. Separation of these steps results in a better accuracy of the test as against the one-step procedure. High sensitivity of the method (the detection limit being 1 mM of cholesterol in a luminometer cuvette) permits high dilutions of the serum, this essentially eliminating the admixture effects on the enzymatic reactions. The findings of the suggested method are in good correlation with those of the spectrophotometric assay of cholesterol using the KONE kit.
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PMID:[The chemiluminescent determination of cholesterol]. 172 47

A manual enzymatic method is described for sensitive fluorometric determination of uric acid in human serum. This method is based on an enzymatic reaction with uricase to form hydrogen peroxide from uric acid and the following oxidation of o-phenylenediamine with peroxidase and hydrogen peroxide for the production of a fluorescence compound. The specificity and the selectivity in the method are due to the uricase reaction and the fluorometry, respectively. The formed fluorescence in the reaction mixture is measured at 410 nm (an excitation) and 550 nm (an emission). This enzymatic method can determine uric acid at 30-1000 microM, with a between-assay relative standard deviation of 4.35% or less. A good correlation is obtained between the present method and the colorimetric kit method.
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PMID:Use of the o-phenylenediamine fluorescence system in the enzymatic assay of serum uric acid. 180 95

A method of "sandwich" enzyme immunoassay was developed for determination of human serum myoglobin with the use of myoglobin isolated from human myocardium and gammaglobulin fraction of a specific sheep antiserum labelled with horseradish peroxidase. The linear part of the calibration curve within the range of 0.08-2.2 nmol/l is suitable for accurate quantitative reading of myoglobin concentration. Intra- and interassay variation coefficients are 7% and 11.2%, respectively. A comparison of 100 serum samples assessed by means of commercially available RIA kit and by the given method revealed a correlation coefficient of 0.86.
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PMID:Enzyme immunoassay to determine serum myoglobin in patients with acute myocardial infarction. 181 Jun 99

MoAbF9 immunoreactivity was investigated in frozen sections of 123 breast carcinomas using an avidin or streptavidin biotin peroxidase kit. A standardized computer image analysis system was used to evaluate immunostaining. The percent of cell surface staining and mean optical densities were correlated with morphological criteria of prognosis such as tumor size histological grade, blood and lymph invasion and axillary lymph node involvement, with immunoreactivity to other MoAb, i.e. Ki67, anti-RE and anti-RP, anti-p.HER-2/neu and with tumor aneuploidy and AgNORs content in tumor cell nuclei. Despite some heterogeneity, MoAbF9 was reactive with all breast carcinomas tested. The percent of F9 immunostained cell surface and mean optical density increased with Ki67 immunoreactivity, tumor aneuploidy and AgNORs nucleus surface but were independent of p.HER-2/neu oncoprotein distribution and tumor receptor content. These findings suggest that F9 could not only allow detection axillary lymph node micrometastases but also be used as plasmatic marker for tumor recurrence and metastases.
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PMID:Monoclonal 3C6F9 distribution in human breast carcinomas: image cytometry of immunocytochemical assays. 182 Apr 90

A rapid, simple dot immunoperoxidase assay (DIPA) is described for visual detection and identification of bluetongue virus (BTV) antigens in samples of infected cell culture fluid. The assay was performed using nitrocellulose (NC) paper and 'dipsticks'. Dots of samples were adsorbed to the NC surface and the remaining non-specific binding sites were blocked with skim milk solution. BTV was detected with either of two murine monoclonal antibodies (4H4, 5G12) to the major group specific antigens of BTV, and the complex was reacted with a peroxidase conjugated anti-mouse immunoglobulin G (heavy- and light-chain specific). Positive reactions were easily visualized as brown spots after enzyme degradation of substrate containing H2O2 and diaminobenzidine (DAB). The DIPA was specific in detecting BTV in samples of cell culture fluid from baby hamster kidney (BHK-21) cells infected with U.S.A. isolates of the five BTV serotypes (2, 10, 11, 13 and 17) known to exist in the U.S.A., and South African isolates of 17 BTV serotypes (1-12, 14-16, 18 and 20), but not with two North American isolates of epizootic hemorrhagic disease of deer virus (EHDV) representing serotypes 1 and 2. Attempts to detect BTV directly in infected sheep blood cells and chick embryo tissue suspensions by DIPA were unsuccessful. Of 55 cell culture fluid samples examined from BHK-21 or Vero cell monolayers inoculated with 55 clinical specimens, propagated initially in embryonating chicken egg (ECE) 11 proved positive and 44 were negative by DIPA. The results were in complete agreement with the conventional ECE and tissue culture isolation systems. The DIPA appears to have potential application, especially as a 'dipstick' kit, for rapid and inexpensive laboratory diagnosis of bluetongue virus infection.
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PMID:Dot immunoperoxidase assay using monoclonal antibody for detection of bluetongue virus antigens. 184 12

For the program to eliminate tetanus neonatorum in this country, we have improved the sensitivity of enzyme immunoassay kit (EIA kit) prepared on 1987 for the detection of human tetanus immunoglobulin (TIG) by competitive principle. (Chinese J Microbiol Immunol 1987; 20: 269-278) Horse-radish peroxidase-conjugated to tetanus toxoid monoclonal antibody was involved in the new kit, and tetanus hybridoma clones were prepared by this laboratory. The lowest detectable TIG level is 0.05 IU/ml serum instead of 0.1 IU/ml serum. The dose-response curve and cut-off determination system of the new EIA kit are better than those of the original one. We proposed that the newly designed EIA kit could be used for understanding the TIG level in women who are in the age group for giving birth and in the tetanus vaccination group.
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PMID:[Improvement of the EIA kit for the detection of tetanus immunoglobulin in human sera]. 185 3


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