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Isolation of serovar-specific leptospiral antigens for use in an enzyme-linked immunosorbent assay (ELISA) compared with the microscopic agglutination test and immunofluorescence.

A total of 10 different detergents were used with the intention of extracting serovar-specific antigens from eight leptospiral serovars for use in an ELISA. Extractions with 2% sodium taurocholate at 50 degrees C proved to be best suited for this purpose. Taurocholate extracted antigens could be separated in SDS-PAGE and antigenic activities demonstrated in immunoblotting. In comparative studies on sera taken from cattle, pigs, horses and dogs, the self-made ELISA proved to be more sensitive than the microscopic agglutination test (MAT) and the immunofluorescence test (IFT). The IFT gave different results from those obtained using MAT or ELISA.

Agglutination Tests↗

The significance of serum sperm-agglutinating antibodies in men with infertile marriages.

Three hundred consecutive male partners of infertile marriages had serum samples tested for sperm agglutinating and sperm immobilising antibodies, using a micro-technique: 30.3% were found to be positive. Out of a control group of 100 fertile men 10 had antibodies detected (10%) and these at low titre; this difference was significant. The results of these serum tests were found to correlate with the results of seminal plasma tests, the mixed antiglobulin reaction for IgG and sperm cervical mucus contact testing, although all of these tests were less discriminatory with regard to fertility. The serum micro-immobilisation test showed a statistical association with fertility when analysed in those couples where both husband's and wife's tests showed no other bar to fertility.

Antibodies↗

Rapid detection of urinary albumin at low concentration by an agglutination inhibition techniqué.

Subclinical elevation of urinary albumin excretion (microalbuminuria) identifies individuals at high risk of developing nephropathy in insulin-dependent diabetes. We have evaluated the performance of a new specific method for the rapid detection of microalbuminuria employing a latex agglutination inhibition technique. A total of 96 consecutive sterile first morning urine samples from insulin-dependent diabetic subjects were tested using AlbuScreen TM (Cambridge Life Sciences). Fifteen samples with albumin concentration greater than 27 micrograms/ml (range 27.6-780 micrograms/ml) read positive with AlbuScreen and were identified with 100% sensitivity and specificity. This test which is ten times more sensitive than the qualitative methods in current use and which detects a wide range of urinary albumin concentrations from microalbuminuria to heavier proteinuria may prove useful in the outpatients setting.

Adolescent↗

Monoclonal antibody inducing human sperm agglutination.

A mouse fused splenic cell line producing antibody that induces agglutination of human sperm was cultivated. The antibody was localized to the midpiece and tail of human spermatozoa by an indirect immunofluorescence method. The interacting antigen was identified in a sperm membrane protein preparation by SDS gel/protein blot immunofluorescence method. The molecular weight of the antigen was in the vicinity of 84 Kdaltons.

Animals↗

Antisperm antibodies detected by ZER enzyme-linked immunosorbent assay kit are not those detected by tray agglutination test.

Antisperm antibodies may play a role in the pathogenesis of infertility, particularly in the male. One of the standardized methods for detecting antisperm antibodies is the tray agglutination test (TAT). Unfortunately, this assay requires fresh motile spermatozoa. Tests for binding of antibody to fixed sperm or sperm extracts have been developed as enzyme-linked immunosorbent assays (ELISA), and we compared the results of using one such ELISA method with the TAT to detect antisperm antibodies in a panel of known positive and negative sera from infertile and control patients. With respect to the TAT assay, the ELISA gave a 75% false-positive test rate and a 63% false-negative rate. It is important to validate new assays such as the ELISA before widespread application to patient screening particularly since patients judged to have antisperm antibodies may be treated with high-dose corticosteroid drugs that are not without significant side effects.

Antibodies↗

Studies on mycobacteria isolated from animals, with special reference to the agglutination test.

Ninety-three strains of slowly-growing mycobacteria were studied biochemically. Ninety of these were isolated from animals (pigs, cattle, dog and poultry) and three from dust and sawdust-bedding in a pighouse. One strain from a lymph node of a pig was identified as M. gordonae. Ninety-two strains fitted into the M. aviam-intracellulare complex. Of the 92 biochemically confirmed M. avium-intracellulare strains, 78 were tested serologically ad modum Schaefer. Of 73 strains from pigs, one was serotype 1, fifty serotype 2 and eight serotype 8, while two could not be type and twelve were autoagglutinable. Three strains from pighouse environment were serotype 8 and two from cattle and a dog were both serotype 2. A slight modification of Schaefer's agglutination method, using smaller amounts of antigen and antiserum, was developed.

Agglutination Tests↗

Laboratory identification of pathogenic Neisseria with special regard to atypical strains: an evaluation of sugar degradation, immunofluorescence and co-agglutination tests.

Sugar degradation tests (SDT) were compared with immunofluorescence (IFL) and co-agglutination (COA) tests for the diagnosis of Neisseria gonorrhoeae (GC) and Neisseria meningitidis (MC). Somewhat more than 5% of the GC strains and 8% of the MC strains were misinterpreted by SDT. On most occasions the disagreement between SDT and serological tests was due to the inability of the MC strains (less so for GC strains) to degrade sugars correctly. Because of this, three out of 15 strains (20%) from pharyngeal specimens were primarily considered to be GC by SDT but were identified as MC by COA tests. Deficiencies in sugar degradations were also found in a group of clinical problem strains. Many of them were unable or had a decreased ability to degrade glucose or maltose but were diagnosed distinctly as MC by the COA test. There were no false positives with the IFL or COA tests, but 2% of the GC strains and 26% of the MC carrier strains (non-groupable) were not identified by COA. Both IFL and COA tests are good adjuncts to SDT for the diagnosis of GC and clinically significant MC, since the results are reliable and the tests rapid and simple to perform.

Agglutination Tests↗

Serology of Neisseria gonorrhoeae. Demonstration of strain-specific antigens by immunoelectrophoresis, immunofluorescence and co-agglutination techniques.

Among a group of four selected strains of Neisseria gonorrhaeae, common and strain-specific antigens were demonstrated by immunoelectrophoresis, indirect immunofluorescence (IFL) and co-agglutination (COA). In preparations of ultrasonically-disrupted organisms, the strain-specific antigens appeared in crossed-line immunoelectrophoresis (C-LIE) with antigen containing intermediate agar gel as a two-peak precipitin line, one peak close to the antigen well, the other towards the anode. A similar pattern was found for each of the four strains studied. These strain-specific precipitin lines were also identified in rocket-line immunoelectrophoresis (R-LIE) tests, which were found useful for comparative studies. Using whole cells, strain-specific antigens were also demonstrated by COA and IFL tests with the use of cross-absorbed antibodies. The results corresponded to those obtained with C-LIE and R-LIE. Immunosorption experiments indicated identity between the strain-specific antigens shown by COA tests and those demonstrated by C-LIE and R-LIE tests.

Agglutination Tests↗

Serology of Neisseria gonorrhoeae. Classification by co-agglutination.

The co-agglutination (COA) method has been adapted for serological classification of Neisseria gonorrhoeae. COA reagents were prepared with selectively absorbed rabbit hyperimmune antibodies against gonoccal (GC) major outer membrane protein (MOMP) serotype strains. Using these reagents, the 16 MOMP reference strains could be referred to at least three antigen classes, tentatively named W, J and M. The GC antigens of class W were divided into three groups I, II and III, and they were in part sensitive to pronase. The antigens of class J reflected strain specific or serotype reactions, some sensitive and others resistant to proteolytic enzymes. The antigens of class M were sensitive to periodate and resistant to pronase. Strains used in serological studies by other authors were tested. The properties of class W correlated well with those of the so-called micro-immunofluorescence and immunotype systems, and class M with those of the so-called endotoxin and acid polysaccharide systems. Strains from three different laboratories could all be grouped by class W and M reagents. Identical strains obtained independently from different laboratories gave very similar reaction patterns with the reagents available. Repeated GC-isolates from patients infected with beta-lactamase producing strains showed stable reactions with class W and J reagents, while there was a time-related variation of the class M pattern. We have found that the COA method is rapid, easy and reproducible in the serological classification of Neisseria gonorrhoeae and all the 117 GC-strains tested could be classified.

Agglutination Tests↗

Comparison of a micro-agglutination test and the indirect immunofluorescence test for Legionella antibodies in patients.

A micro-agglutination (MA) test for antibodies to 10 different Legionella antigens was compared with the indirect immunofluorescence (IF) test. The antigens used were prepared from Legionella (L.) pneumophila serogroups (SG) 1-6, L. micdadei, L.bozemanii, L.dumoffii, and L.gormanii. Sera from 728 patients suspected of suffering from Legionella pneumonia were investigated by both tests. Thirty-five patients showed a significant rise in IF titre to greater than or equal to 128, and 43 had single or static titres of greater than or equal to 256 while the remaining 150 had negative IF titres. The results of the comparison indicate that the MA test offers at least the same sensitivity as the IF test in demonstrating diagnostic titre rises. It may be used as a screening test for selection of sera for further analysis by the IF test. In contrast to the IF test, MA is simple and inexpensive.

Agglutination Tests↗

A comparison of commercial Mycoplasma gallisepticum antigens in the rapid serum agglutination test.

Twenty-three chickens, aged 9 weeks, were housed in contact with a similar group of chickens which were infected with M. gallisepticum. The onset and development of the serum agglutinin responses which were detected by 3 commercial antigens using rapid serum agglutination tests up to 20 weeks of age, differed greatly both between the infected and in-contact groups and, within the latter group, between the antigens. Few reactions, which were weak and transient, were detected in 2 similar groups of chickens, neither infected with M. gallisepticum but 1 vaccinated at 12 and 17 weeks of age with killed Pasteurella multocida oil adjuvant vaccine.

Agglutination Tests↗

The serological diagnosis of bovine brucellosis: an evaluation of the complement fixation, serum agglutination and rose bengal tests.

In a collaborative investigation in which 4 laboratories took part, the Rose Bengal Test (RBT), Serum Agglutination Test (SAT) and 4 different Complement Fixation (CFT) techniques were evaluated in selected cattle for the diagnosis of bovine brucellosis, by comparing the results they gave with the bacteriological examination of a selection of lymph nodes taken from the same animals at slaughter. The RBT correctly classified all but 1 of 79 culture-positive cattle, but was more often positive in culture-negative animals than the other tests. The RBT may be most useful as a screen test. 11% of the culture-positive cattle had SAT titres below 100 iu and almost 4% of them had less than 30 iu. However, the SAT was more effective in cattle vaccinated with the 45/20 vaccine. The CFT, in 1 of 4 techniques used, identified all of the culture-positive cattle at a serum dilution of 1/4 or above and was considered to be far superior to the SAT as a diagnostic test, except perhaps in cattle vaccinated with the 45/20 vaccine.

Agglutination Tests↗

A latex cryptococcal antigen agglutination test for diagnosis and monitoring of therapy for cryptococcosis.

A latex cryptococcal antigen agglutination test (LCAT) was performed on sera obtained during the first 14 days of treatment from 58 animals (46 cats, 9 dogs, 2 koalas and 1 long billed corella) with cryptococcosis. The same commercial kit was used for all samples, and most serum samples were treated with pronase before testing. Sera from all 58 cases tested positive with the qualitative LCAT protocol (using undiluted sera), while sera from all 26 cats without cryptococcosis tested negative. Titres determined using the quantitative protocol ranged from 1 to > or = 131 072 (median titre between 2048 and 4096), with 57 of 58 cases (including all 8 animals that presented for neurological signs) having titres > or = 2 and thus considered positive according to the manufacturer's recommendations. The LCAT titre was positively correlated with disease severity (r = 0.4169; P = 0.0011), and patients with disseminated skin and/or lymph node involvement had significantly higher titres than those that did not (P = 0.0157). The presence of neurological signs, the species of the patient, concurrent viral disease (in cats) and the biotype of the isolate had no significant association with the LCAT titre. Cats that died of active cryptococcosis despite treatment did not have significantly higher titres (P = 0.3010) than those that responded to treatment. Sequential LCAT determinations obtained in 37 patients during treatment provided a useful quantitative indication of clinical progress, although the decline in titre lagged somewhat behind clinical improvement. Generally, the antigen titre declined by 2 to 4 fold per month during successful therapy. Although there are insufficient data to make unequivocal recommendations, we suggest that either antifungal therapy be continued until the LCAT titre declines to less than 1, or therapy be discontinued after a 32 fold or greater reduction in titre, with periodic monitoring of the serum antigen titre. Treatment of serum samples with pronase substantially increased the sensitivity of the LCAT.

Agglutination Tests↗

Prevalence of latent cases of Babesia equi infection in some parts of North West India as measured by the capillary agglutination test.

The prevalence of Babesia equi infection in north west India was assessed by means of the capillary tube agglutination (CA) test. The particulate antigen used in the test was potent and no cross reaction with other related haemaprotozoa was observed. The serological survey showed that from 323 horses from 3 localities there was an overall incidence of 50.1 per cent. In Haryana the incidence was 38.3 per cent in the 196 horses tested, in Uttar Pradesh it was 47.2 per cent from 72 animals and in Rajasthan it was 96.4 per cent from 55 horses.

Agglutination Tests↗

Comparative distribution of the serotypes of Enterococcus faecalis isolated from the urine of patients with urinary tract infections and the feces of healthy persons as determined by the slide agglutination reaction.

In 1992, we reported a new scheme for identifying 21 distinct serovars of E. faecalis by the bacterial agglutination reaction. In this study we isolated strains of E. faecalis from the urine of patients with urinary tract infections and serotyped. We also serotyped strains of E. faecalis isolated from the faces of healthy persons. The results showed that serovars 2, 7, 1 and 3 were the major serotypes a distribution on strains of E. faecalis isolated from patients. Serovars 2, 1 and 13 were the major serotypes of the fecal strains isolated from healthy persons. Single serovars were typable of all strains of E. faecalis isolated from patients in 80.6% (395/496) and in 62.4% of the fecal strains. Examination of the hemolytic zone around colonies on horse/sheep blood agar plates showed that beta-hemolytic strains on horse blood agar plate isolated from patients had 36.1% for all isolated strains from patients and 8.3% for fecal strains isolated from healthy persons.

Agglutination Tests↗

[Measles antibody followed after measles vaccination: comparison with gelatin beads agglutination (PA) antibody, hemagglutination inhibition (HI) antibody and IgG-avidity].

The immunity status of 37 cases has been tested at 6 weeks and 3.5 years after vaccination by using gelatin beads agglutination (PA), hemagglutination inhibition (HI) and avidity of specific IgG. The geometric mean titer (GMT) of HI antibody was 34.5 at 6 weeks and had fallen to 17.9 over 3.5 years. On the other hand, GMT of PA antibody was 36.5 at 6 weeks and had increased to 286.0 over 3.5 years. Also, the average of measles specific IgG avidity was 4.5% at 6 week and had increased to 45.4% over 3.5 years. The multiple regression analysis was performed in order to investigate the relationship of HI antibody and avidity to PA antibody. The significant involvement of the HI antibody and avidity to PA antibody (standardized partial regression coefficients; 0.612, 0.726) was recognized and the multiple correlation coefficient was 0.880 (p < 0.001). Hence it was strongly suggested that the increase of PA antibody with time could be dependent on that of IgG avidity.

Agglutination↗