[Studies on immunized rabbits with Candida albicans--by lymphocyte blastoid transformation test, agglutination test and skin test (author's transl)].
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Neutralisation tests were done with pertussis immunoglobulins and immunosera in which mice were immunised by intraperitoneal injection and challenged 4 hours later by intracerebral injection. The results of the neutralisation tests were compared with those of the agglutination tests done with the same preparations. For 2 pertussis immunoglobulins a correlation was found between the results of both tests, but generally no correlation exists between the agglutinating activity and the protective value of the preparations. There is a more than 100-fold difference in their relationship. Therefore a neutralisation test should be used for potency testing of pertussis immunoglobulins.
An antigen extracted and purified from Neisseria gonorrhoeae B370, type 4, was employed to sensitize charcoal particles for use in an agglutination assay of human sera for antibodies that indicate active gonorrheal infection. The screening card test is more sensitive than the fluorescent gonorrheal test-heated (FGT-H), especially in men. A simple one-step absorption improved the specificity without reducing the sensitivity of the test.
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A slide co-agglutination test (Phadebact Gonococcus Test) for the serological identification of Neisseria gonorrhoeae was assessed on gonococcal-like, oxidase positive colonies from 120 cultures, originating from about 6,500 consecutive tonsillo-pharyngeal specimens received at the Neisseria Department, Statens Seruminstitut. The test was performed after subculture on a serum-free medium, since this procedure was found to reduce the number of strains showing inconclusive reactions (pseudo co-agglutination). If this pseudo co-agglutination does occur, however, the test can be repeated with the addition of trypsin to the test system. This causes the previously inconclusive reactions to be reverted to clearly positive reactions in the case of gonococci, and to clearly negative reactions in more than half of the previously inconclusive reactions with other bacterial strains. The results obtained by the Phadebact Gonococcus Test were compared with those obtained by bacteriological identification procedures. Fifty-six of the 120 cultures examined contained gonococci, and all strains were identified by the slide co-agglutination test (five strains with the addition of trypsin). The remaining 64 cultures were negative or exhibited consistently pseudo co-agglutination (eight strains). The specificity and sensitivity of the reagent was further confirmed by the examination of 53 strains of Neisseria gonorrhoeae and 50 strains representing Neisseria species commonly occurring in tonsillo-pharyngeal specimens. The Phadebact Gonococcus Test was considered to be a reliable alternative to routine bacteriological identification of Neisseria gonorrhoeae.
The latex agglutination test for FDP is widely employed clinically to aid in the diagnosis of DIC and other conditions. Of sera containing RF, 93% demonstrated positive FDP latex agglutination tests. Reducing agents in all instances destroyed the RF agglutinating capability. Futhermore, 86% of sera positive for FDP and RF became FDP-negative following reduction. Therefore RF was responsible for false-positive FDP latex agglutination tests in the majority of patients. Reduction of patient sera is a rapid, simple method to distinguish a positive FDP test from a false-positive due to RF.
The agglutination of avian red cells, particularly pigeon and budgerigar cells, by sera from patients who had been exposed to birds and had respiratory symptoms, as well as from healthy bird fanciers, has been used to detect the presence of avian specific antibody. Agglutination tests performed by standard techniques using bird erythrocytes can be read within 45 minutes. One hundred and sixty-five sera were tested, and the 44 which were precipitin positive gave a positive agglutination test with one exception. Of 61 sera which gave positive agglutination tests, 43 gave positive precipitin tests with unconcentrated serum and seven others after fourfold concentration of the serum. Avian antibodies were demonstrated in the remaining 11 sera by radioimmunoelectrophoretic tests. Absorption of the sera with pigeon IgG and IgM and the use of animal antisera directed against pigeon IgG showed that the agglutination reactions were mediated by antibodies directed against pigeon IgG and IgM, and against light chains derived from pigeon IgG.
A simplified tube O-agglutination test was developed and evaluated for the determination of somatic serogroup (O) antigens of Serratia marcescens. Use was made of Tryptic Soy broth (TSB)-grown O-cells that had been boiled for 1 hour; 0.145 M NaCl proved a satisfactory diluent. Various technical parameters of this test were examined as well. Rabbit anti-O immune sera, that had been elicited with 5 x concentrated, TSB-grown, 1 hour-boiled O-cells of all 15 currently employed O-antigen reference strains of S. marcescens yielded satisfactory O-agglutinin titers. The tube O-agglutination test compared favorably with the indirect hemagglutination technic, although the latter technic yielded significantly higher O-agglutinin titers with merely 7 of the 15 O-antigens of S. marcescens. The tube O-agglutination test permitted detection of higher O-agglutinin titers than a microtiter O-agglutination test utilizing O-cells that had been stained with safranin O. Conversely, titers obtained with TTC-stained O-cells in a microtiter agglutination procedure approximated those yielded by the tube O-agglutination test, but O-cells of the various S. marcescens strains were stained nonuniformly by triphenyltetrazolium chloride (TTC). As before, there existed marked serologic cross-reactivity between O-antigens O6 and O14. A new O6 candidate strain, S. marcescens isolate S 1i, serotype O6:H20, was proposed. Contrary to O-agglutinins of human control serum, the O-agglutinins of rabbit anti-O immune sera proved refractory to treatment with 0.1 M of 2-mercaptoethanol (2-ME) and 0.01 M dithiothreitol (DTT) respectively. Dual absorptions of rabbit anti-O immune sera with killed cells of Staphylococcus aureus Cowan I (protein A), failed to significantly reduce O-agglutinin titers, although human IgG and IgM was bound by protein A. It was tentatively concluded that the 2-ME- and DTT-refractory rabbit anti-S. marcescens O-agglutinins resided in the IgM immunoglobulin class.
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A mixed agglutination (MA) test employing staphylococcal protein A (SpA) instead of an antiglobulin reagent has been developed for the detection of IgG antibodies bound to cell-surface antigens. In the SpA MA test, a single indicator system may be used for the detection of IgG antibodies from several mammalian species. The sensitivity of this test compares favorably with that of the conventional MA test in the detection of most mammalian IgG. The main advantage of the conventional SpA MA test is its usefulness in the study of antigens on cells because of adherence of antibody-coated indicator erythrocytes.
This study presents the results of a number of serological tests, in particular the plate agglutination test using acid antigen stained with rose bengal, in the examination of 440 human sera for brucellosis. In addition, the sera were examined for the presence of specific antibodies of various classes (IgM and IgG). These investigations showed the rose bengal plate agglutination test to be highly specific and sensitive as a rapid method for the diagnosis of human brucellosis.
Most samples of cerebrospinal fluid from 49 patients with or without coccidioidomycosis yielded positive agglutination with the Hyland ("CM") latex-particle agglutination test. Furthermore, simple dilution of sera from patients with or without coccidioidomycosis yielded a strong agglutination reaction with the latex particle agglutination test reagent. These reactions were found with three lots of the latex-particle agglutination test reagent. One lot latex particles was found to give agglutination reactions merely by addition of saline solution or water, though a second lot did not. It is advisable, therefore, that the latex-particle agglutination test not be applied to cerebrospinal fluid, and that it be applied cautiously, particularly with diluted sera.
A microtiter agglutination test (MTA) was developed for assaying anti-Pasteurella multocida antibodies in turkeys vaccinated with live avirulent Clemson University (CU) strain. Safranine-O was incorporated with the decapsulated P. multocida antigen and greaterly enhanced the visibility of the sedimented free antigen, which formed a compact red button on the bottom of the well. The advantages of the MTA test over the tube agglutination test were that: 1) less antigen and serum were required; 2) delineation of the end point was improved by staining the antigen with safranine-O; and 3) less labor was required.
Serological classification of Pseudomonas aeruginosa by the slide agglutination test with live organisms was studied, based on the O antigen schema adopted by the international expert panel sponsored by the Subcommittee on Pseudomonas and Related Organisms of the International Committee on Systematic Bacteriology. The typing results obtained by the slide test with well-absorbed O sera were identical to those obtained by the conventional tube agglutination test with autoclaved organisms. Most O antigens occur singly; but O2, O5, and O16 occur in four combinations. Antigens O13 and O14 are closely related, as are O7 and O8, and it would be convenient to classify organisms possessing these antigens collectively as O7,8 and O13,14.
The tuberculosis agglutination titre as described by Nicholls (1975) was measured without prior knowledge of the diagnosis in 27 patients with open pulmonary tuberculosis, 100 patients with other diseases and 14 control patients with no apparent disease. Reproducibility of the test, as judged by repeat estimations on split samples, was poor. When attempting to distinguish between the disease categories the test gave 29% false negatives in patients with tuberculosis and 52% false positives in patients with other disease. Therefore the tuberculosis agglutination titre cannot be recommended for use in distinguishing tuberculosis from other conditions.
Spontaneous agglutination of Haemophilus gallinarum (HG) cells cultured in Kato's broth was inhibited by trypsin treatment. The treated cell had the same antigenicity as the common antigen prepared from chicken meat infusion broth. Trypsinized antigen could be used in the slide-agglutination test to detect HG antibody in chicken serum.
The degree of agglutination in the tube agglutination test of the sera of 373 "reactor" cattle was compared to that of 800 "non-reactor" cattle from multiple reactor herds in southwestern Ontario. Vaccinated cattle had lower titers than did nonvaccinated cattle. Thus, vaccinated cattle were less likely to be classified as positive than nonvaccinated cattle irrespective of the method of interpreting the results of the tube test. When the full allowance for vaccination was used, vaccinated cattle were less likely to be classified as suspicious than nonvaccinated cattle. The extent of herd vaccination did not appear to markedly influence the effects of vaccination or the prevalence of reactors. Thus, it appeared that vaccination with strain 19 produced a small protective effect (inferred from the lower titers in vaccinated animals), but that the magnitude of this effect was markedly dependent on the appropriateness of the vaccine allowance for classifying cattle. Further work is needed on this aspect of vaccination with strain 19.