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At least 19 recordsLinked to original sources
PubMed1982

Human tear lysozyme. A comparison of electro-immunodiffusion, radial immunodiffusion and a spectrophotometric assay.

In 116 persons (67 men and 49 woman) the tear fluid of both eyes was collected using paper discs and once more after topical anaesthesia. After weighing the discs and eluting the tear fluid, the lysozyme content was measured using two immunological methods [radial immunodiffusion (RID) and rocket immuno-electrophoresis (RIE)] and a spectrophotometric assay. Standard curves were established with purified human lysozyme and egg-white lysozyme. In addition, albumin was determined in all the samples by rocket immuno-electrophoresis. All lysozyme assays showed a high degree of correlation, the spectrophotometric technique giving significantly lower values (compared with RID and RIE) when human lysozyme was used for standardization. The lysozyme content of the tears did not correlate with the fluid uptake in the discs, but showed a significant decrease with age, which was abolished by topical anaesthesia. In contrast, albumin levels were dependent upon the weight and independent of the age of the patient. It is suggested that the determination of albumin as a "reference protein' might help in comparing the results of different tear sampling techniques used by various groups, which induce variable amounts of reflex tearing.

Adolescent↗
PubMed1976

An aspergillus immunodiffusion test in the early diagnosis of aspergillosis in adult leukemia patients.

An Aspergillus fumigatus immunodiffusion test was performed biweekly for one year on 80 hospitalized patients with acute leukemia to determine if serologic conversion accompanied clinical aspergillosis. A micro-ouchterlony technique with agarose was used. The antigens were prepared from concentrated A. fumigatus culture filtrates and the sera were concentrated 3-fold before testing. Of 80 patients, 10 were proved at autopsy, lung biopsy, or closed space culture to have invasive aspergillosis. Six of 10 patients converted from a negative to a positive immunodiffusion test, whereas a seventh patient's weakly positive test became strongly positive. Three patients with documented aspergillosis did not develop a positive immunodiffusion test. Four of the patients who converted from a negative to a positive test were treated early and successfully with amphotericin B. A fifth patient developed immunodiffusion test antibody late in the course and died despite therapy. A sixth patient died of concomitant mucormycosis despite early therapy. Six additional patients who converted from a negative to a positive immunodiffusion test could not be evaluated because of inadequate documentation of aspergillosis. In severely immunosuppressed patients, our immunodiffusion test proved to be a specific but not always a sensitive test for aspergillosis. In 4 patients, biweekly tests showed conversion associated with invasive aspergillosis, which was diagnosed early and treated successfully.

Acute Disease↗
PubMed1996

Thyroglobulin determined in thyroid fine needle aspiration biopsies by radial immunodiffusion and electroimmunodiffusion.

Radial immunodiffusion and electroimmunodiffusion were used to measure thyroglobulin, the main component of thyroid colloid, in thyroid fine needle aspiration biopsies. A linear relationship was established between precipitation ring diameter and thyroglobulin concentration by radial immunodiffusion (0.5-3.0 g/l), and between "rocket" height and thyroglobulin concentration by electroimmunodiffusion (0.1-2.0 g/l). A nearly complete correlation was observed between the two methods (r = 0.97). In radial immunodiffusion the ring diameter is dependent on time of diffusion and on the antiserum concentration in the agar gel. In this study, the observation time was standardised at 48 h, and the rabbit anti-thyroglobulin serum concentration at 26 ml/l. The intrathyroidal concentration of thyroglobulin was determined by radial immunodiffusion and the thyroid find needle aspiration biopsy of 45 thyroid tumours with different cytological-laboratory- and clinical diagnoses. It was found that in colloid nodules or cysts thyroglobulin is markedly higher than in euthyroid nodular goitre (13.7 +/- 11.9 g/l vs. 1.35 +/- 0.8 g/l, p = 0.005). In conclusion radial immunodiffusion and electroimmunodiffusion are precise, easy to perform, low cost, non polluting methods, which do not require high sample dilution (in contrast, high sample dilution is necessary for measurement of thyroglobulin in thyroid fine needle aspiration biopsy by radial immunodiffusion). Measurement of thyroglobulin in thyroid fine needle aspiration biopsy provides a quantitative estimate of colloid, an important marker in the differential diagnosis of thyroid nodules.

Animals↗
PubMed1979

Diagnosis of Corynebacterium pyogenes infection in pigs by immunodiffusion test with protease antigen.

A protease antigen was prepared from the culture supernatant of Corynebacterium pyogenes by concentrating with a flash evaporator and ultrafiltration. It was adjusted to the concentration of 32 units by the single radial immunodiffusion with a tentative standard serum. In the immunodiffusion test, the antigen of 4 units reacted enough with sera having an antibody titer ranging from 1 to 128. As a result, it was decided that the antigen of 4 units should be used in the immunodiffusion test for the detection of protease antibody. By the immunodiffusion test, protease antibody was demonstrated in about 35% of 443 sera from pigs collected at random. The antibody titer showed the distribution of 2 peaks. The summits of the two peaks were seen at 4 and 32 of antibody titer, respectively. The valley between the two peaks was seen at 16 of titer. From the result, a diagnostic criterion of the immunodiffusion test was decided provisionally as follows: above 16 of antibody titer is positive, 1 to 8 suspect, and less than 1 negative. On the other hand, protease antibody was demonstrated in sera from 13 of 14 pigs carrying abscesses from which C. pyogenes had been isolated. Its titer was 8 (in 2 pigs), 16 (in 1), 32 (in 3), 64 (in 6), and 128 (in 1). From these results, it was proposed that the immunodiffusion test with protease antigen be used for the diagnosis of C. pyogenes infection in pigs.

Animals↗
PubMed1983

Comparison of immunodiffusion and crossed-immunoelectrophoresis in the diagnosis of invasive candidiasis.

A retrospective single blind study was conducted to compare the efficacy of crossed-immunoelectrophoresis using concanavalin A intermediate gel with double immunodiffusion tests in the diagnosis of invasive candidiasis. On the basis of cytoplasmic antibody detection, crossed-immunoelectrophoresis with concanavalin A differentiated reliably between Candida albicans fungemia and invasive candida infection, whereas double immunodiffusion did not. With regard to sensitivity, specificity, and predictive value crossed-immunoelectrophoresis with concanavalin A was superior to double immunodiffusion. Analysis of double immunodiffusion precipitin bands revealed fuzzy and sharp precipitin bands which corresponded to mannan (fuzzy) and cytoplasmic (sharp) antibodies in the crossed-immunoelectrophoresis with concanavalin A assay. Therefore, the finding of sharp bands in the double immunodiffusion procedure supports the diagnosis of invasive candidiasis.

Adolescent↗
PubMed1986

Influence of C3 level on the determination of C3d in plasma and synovial fluid by radial immunodiffusion.

The influence of C3 levels on the determination of C3d in plasma and synovial fluid by radial immunodiffusion was investigated. In the method used, C3 is precipitated by 11% polyethylene glycol (PEG), and C3d is measured in the supernatant. In 51 healthy donors, a weak though significant correlation between C3 and C3d levels was found. The mean concentration of C3d was 1.6% of that in aged serum from healthy donors. So, small amounts of C3 (i.e., 1-2% of the normal plasma level) in the 11% PEG supernatants may contribute significantly to the C3d levels measured. A radioimmunoassay that detects C3, C3b, iC3b and C3c was used to measure C3 levels in the PEG supernatants. In PEG supernatants of 4 plasma samples, 0.3-0.6% of the C3 level in normal plasma was found, whereas in those of 2 synovial fluids much higher levels were found (4-10% of the normal plasma level). When purified 125I-labeled antibodies against C3c were added to the gel of the radial immunodiffusion, C3c antigen was detected in the precipitation rings obtained with all PEG supernatants of plasma samples from patients. Therefore, the quantitative contribution of C3 to the precipitation rings in the C3d radial immunodiffusion was analyzed after the addition of an excess of anti-C3c antibodies to the gel. No effect on the size of the C3d-precipitation rings obtained with plasma samples from patients was observed. However, the C3d precipitation rings obtained with synovial fluids were significantly smaller when the gel used in the radial immunodiffusion contained an excess of anti-C3c antibodies together with the anti-C3d serum. We conclude that it is necessary to add an excess of anti-C3c antibodies to the gel used for the radial immunodiffusion, for the determination of C3d levels in synovial fluid. An antiserum against human C3b, which contains both anti-C3c and anti-C3d antibodies, can be used for this purpose.

Blood Donors↗
PubMed1970

Immunodiffusion analysis of Yaba poxvirus structural and associated antigens.

Yaba poxvirus virions were extracted and purified from Rhesus monkey tumors. A saline-soluble virion fraction (Y-xp), obtained by mechanical fractionation of purified virions with an X-press, contained seven components in acrylamide gel electrophoresis; five of these components were reactive in immunodiffusion with whole virion and Y-xp antisera produced in rabbits and monkeys. The saline-insoluble residue remaining after X-press treatment was hydrolyzed with sodium dodecyl sulfate, urea, and 2-mercaptoethanol (SUM). This fraction, Y-sum, contained five components, four of which were demonstrable by immunodiffusion. There was no evidence of antigenic relationships between Y-xp and Y-sum antigens in immunodiffusion. In acrylamide gel electrophoresis, one Y-xp and one Y-sum component had similar mobilities. Y-xp but not Y-sum antisera contained viral-neutralizing antibodies. Virus-free saline extracts of Yaba tumor prepared with Genetron (YS) were essentially devoid of virion structural antigens. They failed to induce precipitating antibodies for virion antigens, were nonreactive in immunodiffusion with virion antisera, and gave low complement-fixation titers with virion antisera. Yaba virion antigens were recovered from the Genetron tumor sediment by SUM and alkaline hydrolysis. Antisera prepared to YS extracts gave a maximum of 17 precipitin lines in immunodiffusion with YS extracts; none was identified as a virion structural antigen. Saline extracts of tumor prepared without Genetron contained immunogenic amounts of 5 virion antigens and 12 to 14 associated antigens. Animals immunized with infected cell culture extracts (virus-free) formed antibodies to six to seven virion antigens. The implications of using extracts of Yaba poxvirus-infected tissues in complement-fixation tests to measure virion antibodies were discussed.

Animals↗
PubMed1987

The role of isotype and epitope specificity of monoclonal antibody mixtures in immunodiffusion reactions.

Twenty monoclonal antibodies to human alpha-fetoprotein have been characterized in terms of IgG subclass, epitope specificity and immunodiffusion properties. The Ig molecules consisted of six G1, ten G2a and four G2b isotypes. The epitope analysis was conducted by a solid-phase RIA employing 125I-labelled AFP. The RIA analysis resulted in the identification of seven determinants, two of which were specific for one antibody each, while others were specific for two or more antibodies. In Ouchterlony tests with the antibodies, none formed bands of immunoprecipitate when diffused individually against the antigen. Tests with all possible mixtures of pairs of the antibodies (190) resulted in positive immunodiffusion responses with only two mixtures. These each contained antibodies of the 2b isotype and demonstrated distinct epitope specificities. The immunodiffusion of mixtures of three antibodies from a group of 12 that were selected to represent equal numbers of isotypes resulted in 84 positive responses and 136 negative responses, i.e. from a total of 220 mixtures. A striking correlation was noted between positive immunodiffusion tests and composition of antibody isotypes and of epitope specificities of the mixtures. Each mixture was found to contain antibodies of at least one 2b isotype and of different epitope specificities. None of the mixtures that lacked a 2b isotype (56) responded in immunodiffusion tests. Similarly, in instances (46) in which the epitope specificities of the antibodies in the mixture were the same, i.e. duplicated or triplicated, the results were again negative despite the presence of a 2b isotype. The comparison of these studies with similar studies of a smaller group of antibodies to pregnancy-specific beta 1-glycoprotein strongly suggests that the G3 antibody isotype may have an immune precipitation enhancing effect similar to the G2b isotype.

Animals↗
PubMedDate not supplied

Immunodiffusion screening method for detection of motile Salmonella in foods: collaborative study.

A collaborative study was performed to validate the performance of the 1-2 TEST for detection of motile salmonellae in foods. Detection is based on observation of an immobilized band of cells. Twenty-three laboratories participated in the study. The 1-2 TEST (immunodiffusion test) was compared with the standard culture procedure (BAM/AOAC; FDA Bacteriological Analytical Manual) for detection of Salmonella in 6 food types: ground black pepper, soy flour, dried whole egg, milk chocolate, nonfat dry milk, and raw deboned turkey. Uninoculated and inoculated samples were included in each food group analyzed. After the tests on the 6 foods were completed, analysis of the data for turkey and soy flour showed that certain collaborators obtained data inconsistent with the data from the majority of collaborators. No specific method deviations to account for the inconsistencies were reported by those collaborators, so the collaborative testing of these 2 foods was repeated. Analysis of data for pepper, chocolate, nonfat dry milk, dried whole egg, and the second set of soy flour and turkey indicated 96.1% agreement between the BAM/AOAC and immunodiffusion test methods. The false negative rates for the immunodiffusion test and BAM/AOAC methods were 3.6 and 1.7%, respectively. There was no significant difference in the productivity of the immunodiffusion test and BAM/AOAC method at the 5% level for any of the 6 foods. The immunodiffusion screening method has been approved official first action for detection of motile Salmonella in foods.

Animals↗
PubMed1985

[Diagnosis of transmissible gastroenteritis in swine by the agar gel immunodiffusion test].

A transmissive gastroenteritis antigen was obtained from intestinal content of pigs with symptoms of the disease, the presence of which was demonstrated by the agar gel immunodiffusion test. It was shown that the antigen was rough and lacking in purity, but it proved to be sensitive and specific. In the conditions of agar gel immunodiffusion in the presence of a reconvalescent serum the antigen produced a clearly expressed precipitation line for 24 to 48 hours. A correlation was also found with 17 out of 25 reconvalescent sera of pigs, investigated via virus-neutralization and agar gel immunodiffusion. Four sera were positive through virus-neutralization only, and four--through agar gel immunodiffusion, while the controls were negative. The agar gel immunodiffusion test was useful in the demonstration of both antibodies in reconvalescent sera and virus in the intestinal content of pigs either suffering or died of transmissive gastroenteritis.

Animals↗
PubMed1978

Two immunofluorescent methods compared with a radial immunodiffusion method for measurement of serum immunoglobulins.

We found high correlations when the values for immunoglobulins G, A, and M, obtained from the analysis of a group of 50 or more human sera by a measured-time radial immunodiffusion method, were compared to values found by two new commercial immunofluorescent methods. The Fiax and the Immuno-Fluor systems make use of fluorescently labeled antibodies and solid-phase immunoadsorbents and offer simple, precise, and objective methods for immunoglobulin quantitation. The Fiax procedure is less time consuming than is the Immuno-Fluor and gives IgG and IgA concentrations that are in close agreement with those found by radial immunodiffusion and IgM values that are significantly lower than the radial immunodiffusion values. The IgG results determined by the Immuno-Fluor method compare well with the radial immunodiffusion results, uhile the IgA values are significantly higher and IgM values significantly lower than those obtained by radial immunodiffusion.

Evaluation Studies as Topic↗
PubMed1984

Standardization of the equine infectious anemia immunodiffusion test and its application to the control of the disease in the United States.

In 1972 the US Department of Agriculture (USDA) established requirements that horses which had immunodiffusion antibody against equine infectious anemia could not be transported interstate. Forty-two states had regulations requiring that horses have a negative equine infectious anemia immunodiffusion test before movement. In order to standardize immunodiffusion testing, it was stipulated in the 1972 regulations that tests must be performed in approved laboratories. The approved laboratories were required to have personnel trained in the immunodiffusion test procedure, to follow the standard protocol, to use licensed reagents, successfully complete proficiency tests, and to report results to federal or state animal health officials. The number of approved laboratories was 160 in June 1983. The number of immunodiffusion tests performed in the United States increased from 82,777 in 1972 to 793,536 in 1977, and has remained at about that level. The percentage of positive samples has decreased from 3.9 in 1972 to 0.6 in 1982. Due to the laboratory standardization program, the reproducibility of test results has been excellent.

Animals↗