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At least 19 recordsLinked to original sources
PubMed1992

Detection of false seroconversions in cytomegalovirus seronegative platelet donors by serial complement fixation or IgG-specific radioimmunosorbent tests.

A panel of 421 cytomegalovirus (CMV) seronegative-platelet donors was followed using complement-fixation (CFT) and IgG-specific radioimmunosorbent tests (RIST) to detect seroconversion. During 4623 person-months of observation 2452 serum specimens were tested, and the annual rate of seroreactivity detected by RIST was 56%. However, most of the positive RIST results were followed by negative results with later serum samples. CFT seroreactivity was a poor predictor of RIST seroreactivity. The high rate of transient seroreactivity detected by RIST may have reflected the lack of absolute specificity of the test. Alternatively, the results could illustrate the difficulty of determining whether an individual harbours latent CMV infection using antibody tests when some seronegative individuals appear to be latently infected. In the light of our observations recommendations for CMV antibody testing of blood donors are proposed.

Antibodies, Viral↗
PubMed1979

A modified sandwich technique for the measurement of IgE.

The paper radioimmunosorbent test (PRIST) has been described as the method of choice for determining low serum IgE levels; the radioimmunosorbent test (RIST) as the method of choice for determining normal and elevated serum IgE levels. By replacing the 125I-labelled anti-IgE antibody used in the paper radioimmunosorbent test by the 125I-labelled anti-IgE reagent used in radioallergosorbent test (RAST) and by changing the serum dilution and the incubation time, this modified sandwich technique (MST) became comparable to the RIST in the normal and elevated IgE-region and showed results similar to the PRIST in the very low IgE-region. The affinity of the 125I-labelled anti-IgE of the RAST proved to be about 2.6 times higher than the antibody used in the PRIST, which explains the improved results in the normal and the good results in the very low IgE-region. The lowest serum IgE level measurable by this method was as low as 0.05 I.U./ml, as determined in 20 cord sera. The mean IgE level in cord sera was 0.45 I.U./ml (range 0.05--2.63 I.U./ml). The results of this study suggest replacement of the antiserum used in PRIST by the one available for RAST.

Asthma↗
PubMed1978

Reutilization of 125I-labelled anti-IgE antibody and paper discs in PRIST and RAST IgE determination.

In the paper radioimmunosorbent test (PRIST) anti-human IgE coupled paper discs are used for the estimation of total IgE in blood serum: in the radioallergosorbent test (RAST) allergen-coupled paper discs are used for the estimation of specific IgE in blood serum. The bound IgE or specific IgE is quantified by a 125I-labelled anti-human IgE. The non-bound 125I-labelled anti-human IgE can be collected and used in new assay. By a 4-h incubation of the used paper discs with 1 M glycine-HCl buffer (pH 2.7) the IgE-labelled anti-IgE complex can largely be removed. The paper discs treated in this manner can be used in a new assay.

Antibodies, Anti-Idiotypic↗
PubMed1977

Comparison of procedures for measurement of IgE protein in serum and secretions.

Because of conflicting reports in the literature concerning the value of various procedures for measurement of IgE in serum and secretions, we compared four different methods, the radioimmunosorbent test (RIST), the double-antibody radioimmunoassay (RIA), the paper disc immunosorbent test (PRIST), and radial immunodiffusion (RID). The standards used in the assays were tested initially in the double-antibody RIA with the use of the reference from the World Health Organization. The results showed that RID as expected was relatively insensitive and IgE was reliably measured only above approximately 1,000 international units (IU). Moreover, certain sera containing low levels of IgE by the other procedures gave distinct precipitin zones and presumably falsely high levels of IgE protein. Thus RID may yield apparently erroneous results when used as a screening procedure for measurement of IgE levels. Among the other procedures PRIST and the double-antibody RIA showed the best agreement. With serum samples RIST yielded values for IgE in the low level range higher than those given by the PRIST and double-antibody RIA. With breast milk and colostrum, values of IgE between 120 and 690 ng/ml were found by RIST, whereas IgE was not detected by double-antibody RIA and PRIST. No evidence of an inhibitor of IgE was found in breast milk, so that the apparent elevation of IgE in breast milk by the RIST is likely false. These findings confirm prior reports of spurious elevations of IgE with the RIST and indicate the usefulness of the PRIST and double-antibody RIA for the measurement of IgE in sera and secretions.

Colostrum↗
PubMedDate not supplied

Immunoglobulin E in chronic middle ear effusions: conmparison of RIST, PRIST, and RIA techniques.

To investigate the possible role of allergy in otitis media with effusion (OME), the immunoglobulin E (IgE) content of 138 middle ear effusions (MEE) and paired serum samples from patients with chronic otitis media with effusion was determined. The initial 62 paired specimens were assayed for IgE by the radioimmunosorbent test (RIST), while the later 76 paired specimens were assayed for IgE by the paper radioimmunosorbent test (PRIST). When the results obtained by these two techniques were compared, it was noted that the PRIST procedure gave significantly lower IgE values for effusions than the RIST method. When the effusion-to-serum ratios (E/S ratios) were computed from the PRIST data, the E/S ratio was less than one, while RIST data gave an E/S ratio greater than one. The results obtained with the PRIST procedure were confirmed by double antibody radioimmunoassay for IgE. Thus, the PRIST procedure appears to measure the IgE content of MEE more accurately, and the results obtained by this procedure fail to support the concept of allergy as a major causative factor in OME.

Adolescent↗
PubMed1990

Diagnosis of cytomegalovirus infection in cyclosporin-treated renal allograft recipients.

The relative merits of antibody response and virus shedding as markers of cytomegalovirus (CMV) infection among cyclosporin-treated renal allograft recipients were analysed. CMV-specific antibody was assayed by IgG-specific radioimmunosorbent test (RIST) and by complement fixation test (CFT). CMV shedding was assayed by virus isolation and by the rapid test immediate early nuclear antigen detection (IENAD). RIST and CFT detected seroconversion in similar numbers of patients, but the former test was the more sensitive when CMV antibody was sought in pretransplant sera to differentiate primary from recurrent infection. IENAD detected or excluded CMV shedding for more urine specimens than virus isolation (462/515 [90%] vs. 366/515 [71%]), but the reverse applied to saliva specimens (33/57 [58%] vs. 54/57 [95%]). The high specificity of IENAD allowed positive results by IENAD to be accepted when virus isolation failed to provide a result. IENAD was, however, less sensitive than virus isolation even when specimens yielding CMV by IENAD, but no result by virus isolation, were included in the analysis (27/44 [61%] vs. 38/44 [86%]). Assays of both antibody response and virus shedding were required to maximise the diagnosis of recurrent CMV infections, but the former assay detected all primary CMV infections. The diagnostic implications of these results are discussed.

Antibodies, Viral↗
PubMed1986

Quantitative measurement of human immunoglobulin E using monoclonal antibodies to distinct epitopes.

Monoclonal antibodies (MAb) which recognize distinct epitopes on human immunoglobulin E have been used to develop two-site sandwich radio- and enzyme-linked immunoassays for the quantitation of human IgE. In the first step, a purified anti-IgE MAb coated to polyvinyl or polystyrene microtiter plates specifically bound the IgE contained in the samples. In the second step, another anti-IgE MAb (either iodinated or conjugated to beta-galactosidase) directed to a different antigenic determinant was used to estimate the amount of bound IgE. This simple method permitted the determination of IgE concentrations of 10 ng/ml and greater in about 3 h. Coefficients of variation on a single day did not exceed 7.5% for IgE levels, covering a wide range of the standard curve. The values obtained on serum samples showed a good correlation with those obtained using the paper radioimmunosorbent test (PRIST).

Antibodies, Anti-Idiotypic↗
PubMed1991

Detection of immunoglobulins from explant cultures of periapical lesions.

The presence of immunoreactive cells in periapical inflammatory lesions suggests that immune responses participate in the disease process. The purpose of this study was to determine the presence and concentration of immunoglobulins in the supernatant fluids of explant cultures of periapical lesions. Ninety periapical lesions that had been contiguous with the apex of a root were removed and maintained in explant cultures for 96 h. Tissue culture medium was replenished at 24, 48, 72, and 96 h. Double diffusion in agarose assays demonstrated the presence of IgG in 100% of the 24-h supernatant fluids and IgA in 65% of the 24-h supernatant fluids. However, IgM was not detected. Radial immunodiffusion assays were used to detect and quantitate IgG, IgA, and IgM in samples of 24-h supernatant fluids from 90 explant cultures. IgG was the predominant immunoglobulin followed by IgA. A radioimmunosorbent test was used to detect and quantitate IgE in samples of 24-h supernatant fluids from 90 explant cultures. Forty of the 90 supernatant fluids contained measurable IgE. All detected immunoglobulins decreased in concentration in daily supernatant fluids with time (24, 48, 72, and 96 h) in the culture.

Adolescent↗
PubMed1989

Nonspecific immunoglobulin E in aqueous humor: evaluation in uveitis.

The levels of immunoglobulin E (IgE) in aqueous humor (AH) and serum were determined in 74 uveitis patients and 13 control patients with senile cataract. Uveitis patients' levels ranged from less than 0.06 to 460 IU/ml according to the paper radioimmunosorbent test (PRIST; limit of sensitivity, 0.06 IU/ml). Only one control patient had an AH level higher than the sensitivity limit (0.08 IU/ml). Of the five patients with the highest levels in the AH, four had toxocaral retinitis. The ratio of AH/serum to IgE normalized to the albumin concentration was calculated in 25 uveitis patients and was greater than 0.60 in 13 cases, indicating a possible intraocular IgE synthesis.

Adolescent↗
PubMed1976

Immunoglobulins in nasal secretion with special reference to IgE. I. Methodological studies.

Different ways of collecting relatively large volumes of nasal secretion with as physiological a composition as possible were studied. Nasal secretion was collected by the so-called nasal spray washing method from 5 patients with allergic rhinitis due to pollen and 5 healthy persons during a pollen-free season. The collection was performed without any provocation and following nasal provocation with histamine or allergen solution. With the radioimmunosorbent test, in which the lower limit of measurement was 0.1 units/ml, IgE could be quantified in 52 of 60 analysed secretions. IgA, IgG and albumin were demonstrated in all secretions. In the allergic patients, following histamine and allergen provocation, a relative increase in the concentration of IgE and albumin and a significant decrease of the IgA/albumin ratio in nasal secretion was found. In the healthy subjects, such changes in the secretion were observed only after histamine provocation. Calculations also suggested some local production of IgE, but not at all of the same order of magnitude as of IgA.

Adult↗
PubMed1979

Analysis of the serum IgE levels in nonimmunized rats of various strains by a radioimmunoassay.

An assay, the paper radioimmunosorbent test (PRIST) for the measurement of total serum IgE in the rat, that allows the measurement of IgE down to a level of 0.25 ng/ml, is described. With this assay it has been demonstrated that significant differences exist in normal serum IgE levels in non-immunized rats. These differences are strain-dependent, and strains may be classified as to low (less than 50 ng/ml), medium (50--300 ng/ml) or high (greater than 300 ng/ml) serum levels. The serum IgE level is not sex-dependent. Environmental factors have a pronounced influence, optimal housing conditions resulting in low and stable levels while suboptimal conditions result in high levels with great fluctuations. Hybridization experiments suggest a multigenic control, with low IgE strains having a dominating suppressor effect.

Age Factors↗
PubMedDate not supplied

Radioimmunoassay of IgE in middle ear effusions.

Fifty-six sample pairs of middle ear effusions and sera obtained from patients with otitis media with effusion were investigated for The IgE antibody activity against mites, using radioallergosorbent test (RAST) which was recently developed as an in vitro allergy test. The IgE concentrations were also determined by radioimmunosorbent test (RIST). Results showed that the IgE antibody activity against this allergen was found in 5 middle ear effusions and 3 sera. Four of 5 patients with the positive RAST test had nasal allergy or signs suggesting allergy. Theree were two middle ear effusions which seemed to be an allergic fluid. However, findings in the present study do not suggest that middle ear effusions result from the direct allergic reaction induced by mites in the mucous membrane lining the middle ear cavity.

Acute Disease↗
PubMed1991

Diagnosis of nasal allergy to the house dust mite.

Twenty-five patients with perennial rhinitis and a positive skin prick test (SPT) for Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farinae (Df) were submitted to nasal provocation and the radioimmunosorbent test (RAST) for specific IgE-antibodies. We found a significant correlation in the reaction to both allergen extracts for all parameters examined. In addition, there was a significant correlation among the SPT, the RAST and the nasal provocation for Dp and between the SPT and the RAST for Df. In patients with perennial rhinitis we recommend the combination of all three methods to differentiate unspecific rhinitis from an allergic rhinitis. Only the patients with proved allergic rhinitis could benefit from a specific hyposensitisation.

Adult↗
PubMed1975

Measurement of absolute amounts of antigen-specific human IgE by a radioallergosorbent test (RAST) elution technique.

A technique for the absolute quantification of antigen-specific human IgE is described. It employs elution of a calculable amount of antigen-specific IgE from an allergosorbent-antibody complex by means of alkaline pH treatment, followed by measurement of the IgE content of the eluate with a modified radioimmunosorbent test (RIST). With this method IgE antibody directed against the benzylpenicilloyl determinant of penicillin (BPO) was measured quantitatively in sera from seven penicillin allergic patients. IgE specific for ragweed antigen E was measured in sera from 33 ragweed allergic patients. Values obtained for IgE anti-BPO ranged from 19 to 1806 ng/ml and comprised from 1.3 to 27.5% of total serum IgE. Values of IgE anti-antigen E ranged from 9 to 1807 ng/ml, comprising from 3 to 84% of total serum IgE. Excellent correlation (r = 0.99; p less than 0.001) was obtained for both antigen systems between values determined by the RAST elution technique and by simple RAST assay with interpolation from a reference serum of known specific IgE content as determined by the elution technique may be needed only for primary standardization of reference sera.

Allergens↗
PubMed1977

IgE studies on respiratory tract allergies.

The IgE content was determined and specific IgE antibodies were identified in serum samples and nasal secretions of 40 control subjects, 103 patients with nasal allergy, and 37 patients with bronchial asthma using a radioimmunosorbent test an both the serum samples and nasal secretions of allergic patients. Approximately 70% of the serum samples and nasal secretions obtained from patients with positive house dust skin test results were found to contain specific IgE antibodies to mites. The results of this study proved the usefulness of the determination of IgE and specific IgE antibodies in the diagnosis and study of the pathogenesis of respiratory tract allergies. The IgE in nasal secretions was much more closely related to pathogenesis of respiratory allergies than was IgE in serum samples. However, unless more concentrated nasal secretions are assayed, the diagnostic value is limited.

Adolescent↗
PubMed1992

Strong serum inhibition of specific IgE correlated to competing IgG4, revealed by a new methodology in subjects from a S. mansoni endemic area.

A method allowing the immunopurification of human IgE from small volumes of sera with a yield close to 100% (mean = 97.8%; SEM = 0.7) has been developed. The immunopurification eluates were cleared of other class antibodies that could compete with IgE in specific assays. Immunopurification of IgE followed by specific IgE enzyme-linked immunosorbent assay (ELISA) (IMMEL) was then applied to sera of 160 individuals from an area endemic for Schistosoma mansoni. In comparison with radioimmunosorbent test (RAST) and ELISA performed on unfractionated sera, IMMEL provided the highest specific IgE signals. Furthermore, the best correlations between the specific IgE levels and either the specific basophil histamine release levels (r = 0.84; p less than 10(-4) or the anti-S. mansoni skin test values (r = 0.45; p = 10(-4)) were obtained with IMMEL. Measurement of anti-S. mansoni IgE levels in immunopurified fractions and in unfractionated sera of these 160 individuals revealed a strong serum inhibition (geometric means of 98.6% and 96.8% for the adult worms and the larvae, respectively) of the specific IgE reactivity in ELISA. This inhibition was correlated with the anti-adult worm and anti-larval IgG4 levels (r = 0.65; p less than 10(-4) and r = 0.58; p less than 10(-4), respectively). In contrast, this inhibition did not correlate with the specific IgG1, IgG2, IgG3 and IgM levels. Furthermore, the level of specific IgG4 was clearly lower than that of specific IgG1, suggesting that the major contribution of IgG4 in the competition effect is not due to higher levels but rather to a specificity spectrum close to that of the specific IgE. These results support the idea that a specific function of IgG4 in serum might be to control antigen recognition by IgE and consequently, to regulate anaphylactic reactions and IgE-mediated immunity.

Animals↗
PubMed1976

Comparison of serum IgE determination by radioimmunoassay and by single radial immunodiffusion.

Two commercially available methods of total serum IgE determination have been evaluated, viz: the radioimmunosorbent test (RIST) and the assay by single radial immunodiffusion (RID). RIST was found a suitable and rapid method for the wide range of IgE concentrations to be expected in the sera of an allergic population. The RID method has a lower limit of detection of about 1000 I.U./ml; over this value, both techniques provided statistically correlated results. However, for technological reasons RID was considered less suitable for routine application.

Asthma↗