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#26698651   2016/03/08 Save this To Up

New Commercially Available IgG Kits and Time-Resolved Fluorometric IgE Assay for Diagnosis of Allergic Bronchopulmonary Aspergillosis in Patients with Cystic Fibrosis.

Allergic bronchopulmonary aspergillosis (ABPA) is difficult to diagnose; diagnosis relies on clinical, radiological, pathological, and serological criteria. Our aim was to assess the performance of two new commercially available kits and a new in-house assay: an Aspergillus fumigatus enzyme-linked immunosorbent assay (ELISA) IgG kit (Bordier Affinity Products), an Aspergillus Western blotting IgG kit (LDBio Diagnostics), and a new in-house time-resolved fluorometric IgE assay (dissociation-enhanced lanthanide fluorescent immunoassay, or DELFIA) using recombinant proteins from an Aspergillus sp. recently developed by our laboratory for ABPA diagnosis in a retrospective study that included 26 cystic fibrosis patients. Aspergillus fumigatus-specific IgG levels measured by a commercial ELISA kit were in accordance with the level of precipitins currently used in our lab. The ELISA kit could accelerate and help standardize ABPA diagnosis. Aspergillus fumigatus-specific IgE levels measured by ImmunoCAP (Phadia) with A. fumigatus M3 antigen and by DELFIA with a purified protein extract of A. fumigatus were significantly correlated (P < 10(-6)). The results with recombinant antigens glucose-6-phosphate isomerase and mannitol-1-phosphate dehydrogenase were encouraging but must be confirmed with sera from more patients. The DELFIA is an effective tool that can detect specific IgE against more fungal allergens than can be detected with other commercially available tests.

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#12100051   2002/07/08 Save this To Up

Der p 2 isoallergens have different allergenicity, and quantification with 2-site ELISA using monoclonal antibodies is influenced by the isoallergens.

Der p 2 isoallergens have been reported and the possibility of different allergenicity has also been suggested. In addition, the quantification with 2-site ELISA may be affected by the isoallergens.

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#9780829   1998/11/16 Save this To Up

Evaluation of a commercial ELISA test for the detection of allergen-specific IgE antibodies in atopic dogs.

A commercial enzyme-linked immunosorbent assay (ELISA) designed to detect allergen-specific immunoglobulin (Ig)E antibodies were evaluated in 36 atopic dogs and in 12 normal dogs. The test showed a sensitivity of 72.23% and a specificity of 41.6%. Positive and negative predictive values were 76.47 and 35.71% respectively. Correlation between the ELISA kit results and intradermal skin testing varied depending on the allergen and ranged from 47.1 to 80.4%, although positive correlation (i.e. allergens positive in both tests) ranged rom 2.7 to 19.4%. In conclusion, this serological test gave both false positive and false negative results. Sensitivity, specificity and predictive values indicate that this ELISA may not be useful in canine atopy. Although correlation studies were hampered by the impossibility of using the same allergenic extracts, the correlation observed between intradermal and serological testing indicates that results from both tests are not interchangeable.

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#8732236   1996/11/19 Save this To Up

Decreased production of IFN gamma and increased production of IL-6 by cord blood mononuclear cells of newborns with a high risk of allergy.

The underlying mechanisms of elevated IgE level in atopic patients are still obscure, however, extensive efforts have been tried to identify an immunological parameter as a predictor of atopy.

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#9008947   1997/02/20 Save this To Up

[Measurement of total serum IgE by enzyme immunoassay with three commercial reagents].

We measured total serum IgE in 14 patients with allergic diseases and 16 healthy subjects, using three commercial ELISA kits. The correlation of results among the three kits was analyzed using Passing and Bablock regression parameters. Results show that measurements of the different kits do not coincide. One kit shows differences using sera from normal subjects. There is no correlation among kits when using sera from allergic patients. It is concluded that it is not possible to determine exactly the amount of IgE using these kits, specially in subjects with elevated levels.

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#8166941   1994/05/27 Save this To Up

[Evaluation of the CIS Allergen Screen I kit versus the Pharmacia Cap System and skin tests in the diagnosis of respiratory allergy].

This study has given evaluation of a new pneumallergen diagnostic test CIS Allergen Screen I in comparison with Pharmacia Cap System and intradermal skin test. Five allergens (Mite (DPT) D1, Mite (DF) D2, Cat E1, Dog E2, Orchard grass G3) have been studied with in vitro tests (CIS Allergen Screen I Cap System) and the results obtained gave on patient to patient comparison a sensitivity of 91%, a specificity of 100% and on allergen comparison a sensitivity of 84%, a specificity of 99% and an accuracy of 93%. Compared with intradermal skin test for two allergens (Mite (DT) D1 and Orchard grass G3), CIS Allergen Screen I have good results to G3 but less specificity and sensitivity to D1. These results could be to depend on different standardisation between allergen extracts especially for Mite.

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#3631654   1987/10/19 Save this To Up

Performance characteristics of immunoenzymatic allergosorbent testing for total and specific immunoglobulin E.

Paper disc-based solid phase radioimmunoassays are widely used in vitro diagnostic test kits for measuring total IgE (Phadebas PRIST) and specific IgE antibodies (RAST). Recently, these kits have been modified by the substitution of an enzyme-linked immunosorbent assay detection system (Phadezym PRIST/RAST). We studied the performance characteristics of Phadezym PRIST and RAST kits. Phadezym PRIST was sensitive to 0.5 IU/mL IgE. Reproducibility was excellent in the range of 5 to 200 IU/mL IgE and adequate in the range of 5 to 1000 IU/mL using 1:10 serum dilutions (average inter-assay coefficient of variation = 19%). Phadezym RAST was specific, but sensitivity was limited by absorbances in the RAST class 1/0 range indistinguishable from background values. Average inter-assay coefficient of variation was 29% for the semi-quantitative 'Phadezym RAST Unit' (PRU) reporting system. We modified the test kit procedure by disc incubations in microtiter plate wells with rotational agitation and use of ELISA-dedicated spectrophotometer and computer software. These microplate accelerated computerized assays ('MacPRIST' and 'MacRAST') were shown to perform similarly to the conventional Phadezym procedures with advantages in speed, ease, and handling of data.

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#3318239   1988/01/21 Save this To Up

[Use of solid-phase immunoenzyme analysis for determining allergen-specific IgE antibodies].

An ELISA system for the detection of allergen-specific IgE antibodies to ragweed allergen has been developed. The system is highly sensitive and specific. Ragweed pollen allergen has been obtained by the dialysis of water-soluble extract through a kidney membrane. The high molecular fraction of ragweed allergen, showing the whole of the allergenic activity detected by skin tests in untreated patients, has been used for coating polystyrene assay plates. To detect IgE antibodies to ragweed allergen, the conjugate of sheep anti-IgE antibodies with horse-radish peroxidase has been used. The level of allergen-specific IgE antibodies has been determined on the basis of the data on the optical density of the samples in comparison with that of the normal sera. The correlation factor of the results obtained in the assay of specific IgE antibodies with the newly developed assay system and with the commercial kit Phadezyme RAST manufactured by Pharmacia AB (Sweden) has proved to be 0.82 at n = 39, p less than 0.01, while the variation factor in the reproduction of the assay results has proved to be 12% at n = 40.

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#6851184   1983/07/08 Save this To Up

The development of a rapid ELISA for IgE utilizing commercially available reagents.

An enzyme-linked immunosorbent assay (ELISA) for human immunoglobin E (IgE) has been developed. To coat the polystyrene tubes (the solid phase) an antibody concentration of 6 mg/l in sodium carbonate buffer, pH 9.8, at 37 degrees C for 24 h was found superior to other conditions. The maximum amount of globulin adsorbed to the polystyrene surface was estimated to be 2.7 mg X m-2. This is consistent with a monolayer being adsorbed. While some of the anti-IgE molecules are inactivated during adsorption, the remaining molecules, once adsorbed, appear to retain activity for extended periods, and the coated tubes were stable for several weeks. Kinetic studies of the association of analyte, of antibody-enzyme conjugate and enzymic catalysis were used for the optimisation of the assay and allowed us to reduce assay time to 6 h. The association of analyte to the coated tube and the association of anti-IgE-peroxidase conjugate to bound IgE, in the present design, had T1/2 of approximately 0.5 h. Equilibrium is not obtained. The dissociation of analyte, surprisingly, was nearly undetectable. At least one of the two antibodies employed in the sandwich assay, preferably the anti-IgE-peroxidase conjugate, should be free of non-specific antibodies in order to eliminate non-specific reactions. The influence of serum matrix was eliminated by diluting serum samples and standards 1:21 in buffer. For the assay of very low concentrations (i.e. less than 10 kIU/l) standards produced in IgE-free serum are required to compensate the influence of matrix. Comparison with a commercial kit (Hoechst-Behringwerke, Frankfurt/Main, FRG) showed good agreement. All reagents are commercially available and the assay is well suited for routine use.

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#6762821   1983/05/27 Save this To Up

[Quantification of total IgE on microtiter plates by enzyme immunoassay].

This work describes an immunoenzymatic method for the detection of total IgE in human sera. The assay is performed using specially formulated microtiter plates as solid phase (Dynatech microElisa). This facilitates the dispensing of the reagents, the washing steps and the reading of the results. The whole assay is done using commercially available reagents. This guarantees the high specificity and reliability of the test as well as the low interassay variation. The method uses 10 micro liters of human serum and is performed in two work days. IgE levels ranging between 10 and 1000 I.U./ml can be accurately assayed without modifications of the test volumes. The microplate test, when compared with other methods, has an additional advantage: the absorbance values of the wells can be processed by a microcomputer. The method has been compared with a commercial kit (EnzygnostTM) and the correlation found in 150 cases was excellent (r = 0.983). Using this method, we have assayed more than one thousand sera with very satisfactory results. In addition, the cost of each test has been reduced by 75%.

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