Evaluation of a human anti-mouse antibody rapid test for patients requiring radio-immunodiagnostic (original) (raw)
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Detection of Antinuclear Antibodies by Solid-Phase Immunoassays and Immunofluorescence Analysis
Clinical Chemistry, 2004
Background: Antinuclear antibodies (ANAs) are associated with several inflammatory rheumatic diseases. The aim of the present work was to evaluate enzyme immunoassays (EIAs) and compare them with classic immunofluorescent analysis (IFA) for the detection of ANA. Methods: Seven enzyme immunoassays were used in this study. All assays were applied as described by the manufacturers. Three populations were included in the study: (a) a population of patients with well-established autoimmune inflammatory disease (n ؍ 102); (b) a population in which a rheumatic disease was diagnosed up to 5 years after an IFA was performed (n ؍ 164); and (c) a population of consecutive outpatients suspected to have a rheumatic disease (n ؍ 91). The current clinical diagnoses of the patients served as the standard against which performance of the assays was evaluated. Results: In patients with well-established rheumatic disorders, the newly developed EIA in which HEp-2 extracts were included had sensitivities and specificities comparable to or in some instances better than the IFA. The assays without HEp-2 extracts included had significantly lower sensitivities and specificities. In the outpatient population, up to 51% of patients had positive ANA tests that did not correspond to classic ANA-associated disease. However, in the assays in which the HEp-2 extracts were not included, the false-positive rate was <10%. The false-negative rate judged against IFA differed from assay to assay and disease to disease and was mostly <10%. Conclusions: In this study, the sensitivities of EIAs and IFA were largely comparable. However, EIAs without HEp-2 extracts included had a low sensitivity but a high specificity, particularly in nonselected populations. The choice of test is highly dependent on the clinical setting in which the ANA test is to be used and on laboratory policy.
A new method for measuring antibody using radiolabeled protein A1 in a solid-phase radioimmunoassay
Journal of Immunological Methods, 1979
A micro solid-phase radioimmunoassay was developed which utilizes radiolabeled staphylococcal Protein A ([12SI]Pro~ein A)in place of radiolabeled anti-immunoglobulin ([12SI]anti-IgG) for the measurement of antibody. For the assay, antigen is adsorbed to the wells of a microtiter plate followed by dilutions of serum and [12sI]-Protein A in subsequent steps. We found that this assay can be used to measure antibody (Ab) against a variety of antigens in human and rabbit but not goat immune serum. Binding of [12sI]Protein A and [12SI]anti-IgG to human and rabbit IgG was comparable. It was possible to quantify this amount of Ab in human serum by reference to immune rabbit serum. The sensitivity of this assay for rabbit antibody was 1 ng/ml.
1985
A relatively simple, specific and sensitive radioimmunoassay system has been developed for the detection of heterophile Hanganutziu-Deicher (H-D) antigen(s) and antibodies. The 125I-labeled H-D antigen-active molecule used for the assay is a bovine erythrocyte major glycoprotein previously found to have a strong H-D antigen potency. The antigen-antibody complex was precipitated with normal human serum as the carrier protein, followed by the addition of rabbit anti-human IgG F(ab')2 serum. With this method, different H-D antigen-active molecules were compared for heterophile H-D antigen potency with reasonable sensitivity detecting about 0.3 ng of cold glycoprotein. 8 different lung cancer tissues were assayed for H-D antigen. The sera from the 8 lung cancer patients were also screened by ELISA and RIA in an attempt to correlate expression of H-D antigen on tissues with elevation of H-D antibodies. The results showed that all patients' tissues expressed the antigen(s) but only 3 of them had abnormal levels of H-D antibodies. This could have been due to excess antigens in circulation or immune complexes.
Clinical chemistry, 1993
The increasing use of monoclonal antibodies (MAbs) for disease diagnosis and therapy has created a class of patients at risk for systematic error in clinical testing due to interference by human anti-murine antibodies (HAMA). HAMA interference is often difficult to detect and can cause either an increase or a decrease in apparent concentrations of antigen present. We undertook a clinical study to test a HAMA-resistant enzyme immunoassay (EIA) format for carcinoembryonic antigen (CEA) determination. Using the Food and Drug Administration-approved CEA-EIA Monoclonal One-Step Assay (Abbott) with the addition of an acid/heat extraction of patients' specimens, we found that the resulting CEA values accurately reflected the patients' status. We demonstrated that the acid/heat-extracted specimens yield linear dilution curves and show analytical recoveries of added CEA in the range of 76-123% in HAMA-positive specimens and 86-103% in HAMA-negative specimens. The correlation of CEA v...
Radioimmunoprecipitation Assay for Australia Antibody
Vox Sanguinis, 1973
The prozone phenomenon that is observed in the radioimmunoprecipitation (RIP) test at lower dilutions of the sera tested for Australia antibody can be eliminated by adequate dilution of the primary antigen-antiserum mixture after incubation of the latter. Fifty normal persons negative for Australia antibody in the counterimmunoelectrophoresis (CIEP) were found also negative in the RIP test. Out of 50 patients recovering from Australia antigen-positive hepatitis, 2 developed antibody detectable in both tests, RIP and CIEP, 40 were negative, and 8 showed low antibody titers in the RIP test with negative findings in the CIEP. It does not seem possible to evaluate the incidence of serum hepatitis infection within the general population using the RIP test for Australia antibody.
The Diagnostic Role of Radiolabeled Antibodies
2017
In order to image tumors with antibodies, one must target those antigens on the tumor cell that are different, either qualitatively or quantitatively, from antigens on surrounding normal cells. Ideally, the targeted antigens would be unique to tumor cells (ie, not found in any normal tissue in any amount). However, in the real world, most “tumor-associated antigens” are also found in some normal tissues, although sometimes at lower density or in less accessible sites (eg, intracellular) than in tumors. The sensitivity and specificity of radiolabeled antibody imaging depends, in part, on the degree of expression of accessible antigen in the tumor site vs normal tissue [1]. Among the first antigens targeted for immunoscintigraphy were the oncofetal antigens (eg, carcinoembryonic antigen [CEA] and alpha-fetoprotein [AFP]). Later, monoclonal antibody development led to the discovery of other, previously unidentified tumor-associated antigens, (eg, tumor-associated glycoprotein 72 [TAG-7...
Solid phase radioimmunoassays using labelled antibodies: A conceptual framework for designing assays
Journal of Immunological Methods, 1977
A simple theoretical model for the antigen-antibody reaction is presented and used to evaluate the optimum conditions for designing solid phase radioimmunoassays (RIA) using labelled antibodies. Both theoretical and experimental data are presented, using a wide variety of antigens and their corresponding antibodies. The types of RIA described include the direct, the indirect, the direct sandwich assays for detecting either antigen or antibody. The experimental results confirm in a semiquantitative manner that the greatest sensitivity of the RIA is achieved when the smallest amount of labelled antibody is used, that whenever possible the antigen/antibody ratio should be greater than unity (> 1), and that the formation of the antigen-antibody complex is dependent on the mass action effect.
Autoimmune Diseases, 2014
Anti-nuclear antibodies (ANA) have traditionally been evaluated using indirect fluorescence assays (IFA) with HEp-2 cells. Quantitative immunoassays (EIA) have replaced the use of HEp-2 cells in some laboratories. Here, we evaluated ANA in 400 consecutive and unselected routinely referred patients using IFA and automated EIA techniques. The IFA results generated by two independent laboratories were compared with the EIA results from antibodies against double-stranded DNA (dsDNA), from ANA screening, and from tests of the seven included subantigens. The final IFA and EIA results for 386 unique patients were compared. The majority of the results were the same between the two methods ( = 325, 84%); however, 8% ( = 30) yielded equivocal results (equivocal-negative and equivocal-positive) and 8% ( = 31) yielded divergent results (positive-negative). The results showed fairly good agreement, with Cohen's kappa value of 0.30 (95% confidence interval (CI) = 0.14-0.46), which decreased to 0.23 (95% CI = 0.06-0.40) when the results for dsDNA were omitted. The EIA method was less reliable for assessing nuclear and speckled reactivity patterns, whereas the IFA method presented difficulties detecting dsDNA and Ro activity. The automated EIA method was performed in a similar way to the conventional IFA method using HEp-2 cells; thus, automated EIA may be used as a screening test.