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Single-tube mixed agglutination test for the detection of staphylococcal protein A.

A simple, rapid mixed agglutination test using sheep erythrocytes (SRBC) sensitized with rabbit hemolysin and intact viable staphylococci is described for the detection of bound staphylococcal protein A. Soluble protein A was heat extracted from 50 clinical isolates as well as the Cowan I and Wood 46 strains of Staphylococcus aureus and titered by a hemagglutination test using sensitized SRBC and dilutions of soluble protein A. Protein A could be detected in all of these supernatants including that of S. aureus Wood 46, a strain generally considered to be protein A negative. These organisms were later retested by the mixed agglutination test and even those staphylococcal isolates expressing very low heat-extractable soluble protein A concentrations (1:2 titers) were positive, confirming the sensitivity of the test. In a screen of clinical isolates, only 4 of 235 (1.8%) coagulase-positive isolates were negative in the mixed agglutination test. Of 25 coagulase-negative isolates, none yielded a positive reaction.

Agglutination Tests

Latex agglutination tests for measurement of antiplague antibodies.

A latex agglutination test was evaluated as a method for detection and titration of antiplague antibodies. Slide and microtiter techniques using polystyrene latex particles coated with specific fraction-I-antigen of Yersinia pestis were found to be comparable in specificity and sensitivity to serological tests commonly used in laboratory practice. The latex agglutination titers correlated well with those measured by the World Health Organization standard method of indirect hemagglutination, although there was a tendency for the former to be a little lower than the latter. With further study, the latex agglutination test may have application in the seroinvestigation of plague infection in rodents.

Animals

Serological classification of Pseudomonas aeruginosa by a slide agglutination test.

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.

Agglutination Tests

An evaluation of the tuberculosis agglutination test.

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.

Agglutination Tests

Assessment of the agglutination test for tuberculosis.

A new whole cell agglutination test for active tuberculosis, for which encouraging results have recently been reported, has been carried out on the serum of 112 subjects. The results have been disappointing; the test had no predictive value in the diagnosis of active tuberculosis.

Agglutination Tests

The effects of Brucella strain 19 vaccination on titers in the tube agglutination test: a case control study.

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.

Agglutination Tests

Rapid slide agglutination test for Lancefield grouping of streptococci.

A rapid slide agglutination test (the Phadebact [PB] Streptococcus test) was compared with the standard autoclave extraction method of Lancefield and presumptive clinical laboratory tests for grouping of streptococci (bacitracin disk sensitivity for group A and sodium hippurate hydrolysis for group B). Identification of group A streptococci by the PB kit was statistically as accurate as by the Lancefield method, whereas bacitracin grouping was significantly less accurate than the Lancefield method (P = less than .02). With regard to group B, there was no statistically significant difference between the PB test and the sodium hippurate test. The PB test correctly identified all group C and G streptococci. The PB kit provides a rapid and reliable method for Lancefield grouping of streptococci.

Agglutination Tests

The slide agglutination test for the diagnosis of filariasis in camels.

The slide agglutination test was adapted for the diagnosis of filariasis in camels, using an antigen prepared from the microfilariae by a simple lytic technique. The preliminary results were satisfactory as the test detected 86 per cent of the infected animals. Only 6 per cent of the healthy camels with no blood parasites or microfilariae in their blood gave positive results and no positive reactions were obtained from 18 animals suffering from Trypanosoma infection.

Agglutination Tests

Antibodies and the Aberdeen typhoid outbreak of 1964. II. Coombs', complement fixation and fimbrial agglutination tests.

In a previous paper (Brodie, 1977) the value of the Widal test in the diagnosis of typhoid fever was shown to be limited. Evaluation of possible alternative tests showed that: (1) the sensitivity of the anti-human globulin (Coombs') test appeared greater than that of the agglutination test but the length of time (48 h) before results were available rendered it of little value in rapid diagnosis: (2) the complement fixation test offered no particular help towards diagnosis; (3) immunized and non-immunized typhoid fever patients developed fimbrial antibodies, as also did immunized healthy individuals. In this latter group, however, those immunized with alcoholized TAB vaccine had higher antibody titres to fimbrial antigen than those immunized with heat-killed phenolized vaccine.

Agglutination Tests

[A rapid direct latex agglutination test for HCG (author's transl)].

792 urine samples from pregnant patients were investigated by a direct latex agglutination test (LA). Results of this slide test were compared with data derived from a haemagglutination inhibition test (HI). The same results were obtained by both pregnancy tests in 768 (96.7%) out of 792 urine samples. The pregnancy test was negative in 20 cases (2.5%) as assessed by HI, whereas a positive result was recorded with the LA in these cases. Seven were cases of early pregnancy and control tests performed by HI became positive at a later date. The remaining 13 (1.6%) of these patients belonged to a group of pathological pregnancies (missed abortion, threatened abortion, incomplete abortion and ectopic pregnancy). The slide test is more sensitive (1000 I.U. HCG/1 urine) than the HI (1500 I.U.HCG/1 urine). No false positive results were obtained with the LA; false negative results were registered in only 0.5% of cases. A semi-quantitative HCG determination was performed by means of the tube and slide test in 29 urine samples. However, agreement of the data by the two methods was relatively poor, owing to the higher sensitivity of the LA, with consequent inaccurate assessment of HCG excretion. Not much importance need be attached to this finding in view of the diagnostic and prognostic deficiencies of HCG determination. The new slide test was found to be a rapid, simple and accurate pregnancy test.

Agglutination Tests

Rapid isolation and identification of group B streptococci from selective broth medium by slide co-agglutination test.

Direct identification of group B streptococci from a selective broth medium was performed with the Phadebact streptococcus test to determine the feasibility of this technique for early detection of streptococcal colonization. Of 124 clinical isolates, 122 (98.4%) were correctly identified in less than 24 h from the selective broth medium, whereas standard cultures from blood agar plates identified, after 48 h, only 66 (53.2%). The presence of group B streptococci in mixed cultures was always detected by the Phadebact test, and no false-positive co-agglutination tests were observed in 372 cultures from which organisms other than group B streptococci were isolated.

Adult

An agglutination test for the detection of Bordetella bronchiseptica infection in swine.

An agglutination test with the use of formalin-killed antigen of the cell carrying the capsule was developed and used for the detection of antibody in swine naturally infected with Bordetella bronchiseptica. Under optimum antigen concentration and reaction temperature 210 or 60% of 342 serum samples tested from 42 conventional swine herds were positive for Bordetella infection. In contrast, only 34 or 10% of 342 nasal swabs from the same animals were positive for Bordetella by culture technique. The test was relatively free of cross-reactivity to related organism. However, 2.7 and 13.0% of sera from growing pigs and mature hogs, respectively, reacted with antigen of Pasteurella multocida. Because of this, only agglutinin reactions in 1:20 dilutions or higher to Bordetella were considered positive. The bulk of the antibody activity of selected sera tested from various age ranges of swine was mercaptoethanol sensitive, suggesting that serum antibody in Bordetella infection may be associated with immunoglobulin IgM. Because of the high agglutinability and stability of formalin-killed antigen the test may be useful as an auxiliary aid for the diagnosis of Bordetella infection where the organism cannot be identified by culture means.

Agglutination Tests

[Highly specific agglutinating O- and OK-immunoglobulins for identification of Escherichia in the agglutination test on glass].

Escherichia agglutinating O- and OK-immunoglobulins of class G were obtained by the method of ion exchange chromatography; in studying with live and heated O- and K- cultures of the test strains from the International escherichia collection these immunoglobulins proved to be diagnostically highly specific and useful for the identification of escherichia strains in the express agglutination reaction on glass.

Agglutination Tests

A latex agglutination test for rapid quantitative estimation of the plasmin-antipalsmin complex in human plasma.

An antiserum was raised in rabbits against human plasmin-antiplasmin complex and rendered specific for neoantigens of this complex by absorption with purified plasminogen and plasma. Polystyrene particles were coated with the specific antibodies and used in an agglutination test for the determination of plasmin-antiplasmin complex in the plasma from various patients. Purified plasmin-antiplasma complex at a concentration of 0.1-0.2 mg/l was found to cause a clear agglutination of the particles. Activation of fresh human plasma with urokinase caused progressive generation of agglutinating activity up to a plasma dilution of 1/480. Intravenous infusion of streptokinase into patients resulted in an increase of the plasmin-antiplasmin titre of at least 1/240. Sera from patients with rheumatoid factor also agglutinated the particles but this activity could be removed by absorbing rheumatoid factor on insolubilized human IgG. Out of 101 male and twenty-three female control subjects, only three men had a plasmin-antiplasmin titre above 1/16. Of 230 hospitalized patients, plasmin-antiplasmin titres of 1/40 or more were detected in twenty-five patients. Most of these patients had diseases which are frequently associated with in vivo coagulation or fibrinolysis, but among them there was only one who showed diffuse intravascular coagulation detectable by classical methods. In the absence of an increased plasmin-antiplasmin titre none of the haemostasis analyses were indicative of in vivo coagulation or fibrinolysis. Seven out of eight patients with diffuse intravascular coagulation of various origin had plasmin-antiplasmin titres of 1/80 or 1/160. Thus, the present latex agglutination test, owing to its simplicity and sensitivity, appears to be a practical routine screening test for detecting fibrinolytic activation in plasma.

Adolescent

The use of spleen antigen in the tube agglutination test for diagnosis of anaplasmosis in cattle.

An antigen was prepared from the spleen of an infected bovine calf and was used in the tube agglutination test. Cases having rare and very low parasitaemia as well as the carrier cases were detected by this procedure and no non-specific reaction was observed with other haemoprotozoan infections. The maximum titre obtained with the Anplasma positive sera was 1:20, about 60 per cent showed a titre of 1:10 and 90 per cent showed a titre of 1:5 or more. The test provided satisfactory results in detecting the cases even at the early phase of the infection. The test also proved useful in judging the efficacy of a drug employed to destroy the carrier status.

Agglutination Tests