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PubMed1986

Microtiter latex agglutination--quantitative and qualitative equivalent to hemagglutination inhibition for detection of rubella antibody.

Sera were tested for rubella antibody by a standard hemagglutination inhibition method and by latex agglutination in microtiter plates (Virogen Rubella Micro Test). When 157 sera were examined by hemagglutination inhibition and by microtiter latex agglutination for immune status, 149 of the results agreed. The eight discrepant sera (all hemagglutination inhibition-negative, latex agglutination-positive) were tested by a card latex agglutination method (Rubascan) and a passive hemagglutination method (Rubacell). Results, except where noted, agreed with those of the microtiter latex agglutination. One hundred forty-eight of the 149 sera that were in agreement as to immune status were also within a fourfold variation when hemagglutination inhibition and latex agglutination titers were compared. An additional 100 sera were tested for immune status by microtiter latex agglutination and by passive hemagglutination with the single discrepant result agreeing with the latex agglutination when tested by a third method. Both microtiter latex agglutination and hemagglutination inhibition agreed in detection of fourfold or greater rises in each of 20 paired sera.

Antibodies, Viral↗
PubMed2000

Comparison of four serological methods for the detection of diphtheria anti-toxin antibody.

The aim of this study was to compare four serological methods for the detection of Corynebacterium diphtheriae IgG anti-toxin antibodies (IgG-DTAb) in human serum. One hundred serum samples were evaluated for C. diphtheriae IgG-DTAb by four different methods: passive haemagglutination (PHA), latex agglutination test (LA), toxoid enzyme-linked immunosorbent assay (Toxoid-ELISA), and toxin-binding inhibition enzyme-linked immunosorbent assay (ToBI-ELISA). As the external standardisation the neutralisation test for C. diphtheriae toxin in Vero cells (TN Vero) was used. For internal standardisation of IgG-DTAb titres, the WHO standard serum of human diphtheria antitoxin was used. The study revealed a poor correlation between the reference test and the PHA (r=0.34 Pearson's correlation coefficient), an acceptable correlation for the LA (r=0.74), a good correlation for the Toxoid-ELISA (r=0.81) and a very good correlation for ToBI-ELISA (r=0.93). The sensitivity measurements of PHA, LA, Toxoid-ELISA and ToBI-ELISA tests, were 14, 100, 94, 96% respectively and the corresponding specificity characteristics were 86, 76, 94, 90 respectively. Of the four evaluated methods, the ToBI-ELISA could be recommended for scientific and precise laboratory assays of diphtheria antibody levels in humans. For screening purposes the Toxoid-ELISA could be used, but the accuracy of antibody titres below 0.1 IU/ml, considered as the limits of protection, is questionable. Both tests offer very useful alternatives to the in vitro diphtheria toxin neutralisation test in Vero cells. Because of their unsatisfactory correlation and sensitivity as compared to the reference method, PHA and LA should be avoided and replaced by one of the two enzyme immunoassays.

Animals↗
PubMed2003

Double aldehyde stabilisation of erythrocytes in the indirect hemagglutination for echinococcosis.

Sheep erythrocytes were stabilised with glutaraldehyde tanned and fixed with formalin in the indirect hemagglutination test (IHA-GF) and sensitised with hydatid antigen for the diagnosis of human cystic echinococcosis (CE). The sensitivity of this method was compared to that prepared with fresh tanned cells (IHA-TA) in 278 sera from hydatid patients. The sensitivity of IHA-GF (87.8%) was higher than that of IHA-TA (85.6%), the difference being insignificant. Higher geometric mean titres were obtained by IHA-GF (1:13300) than by IHA-TA (1:11600). The use of two sorts of aldehydes proved to be a satisfactory method, showing high sensitivity, a very good specificity and some advantages. The sensitised cells retained their diagnostic effectiveness for at least 15-18 months when stored at 4 degrees C. The technique is inexpensive and rapid, allowing the testing of a large number of sera. The method reduces the variation of the results and improves the reproducibility of the test. When the minimal diagnostic titre-1:400 is used the specificity of IHA-GF might increase by 2.9% while the sensitivity might decline by only 1.4%. The IHA-GF demonstrated better immunodiagnostic characteristics than enzyme linked immunosorbent assay (ELISA) and Latex agglutination test (LAT). The IHA-GF should be considered as an useful method in the range of classical diagnosis for the serology of CE. The clinical diagnostic potential should be increased by a combination of at least two tests: IHA-GF and ELISA or LAT.

Animals↗
PubMed2006

Evaluation of six agglutination tests for Staphylococcus aureus identification depending upon local prevalence of meticillin-resistant S. aureus (MRSA).

Most routine laboratory detection of Staphylococcus aureus isolates is based on rapid agglutination test systems. Failure of agglutination assays to identify meticillin-resistant S. aureus strains (MRSA) has been demonstrated. The aim of this study was to evaluate six commercially available agglutination tests for the detection of meticillin-sensitive S. aureus (MSSA) and mecA-positive MRSA strains. The Dry Spot Staphytect Plus test (Oxoid), the Pastorex Staph Plus test (Bio-Rad), the Slidex Staph-Kit and Slidex Staph Plus test (bioMérieux), the Staphaurex Plus test (Remel) and the Staphylase Test (Oxoid) were used. As determined by pulsed field gel electrophoresis, 52 distinct MRSA strains from five countries, 83 MSSA strains and 150 coagulase-negative staphylococci were included. Species identification and determination of susceptibility patterns were performed using colony morphology, Gram stain, catalase testing, tube coagulase testing, DNase testing, mannitol fermentation, susceptibility testing towards oxacillin by Etest, coagulase gene PCR, fibrinogen receptor gene PCR and PCR of the mecA gene. Sensitivity of the agglutination tests ranged from 82.7 to 100.0 % for MRSA strains and 92.8 to 100.0 % for MSSA strains, respectively. Specificity of the test systems ranged from 91.3 to 99.1 %. None of the six agglutination assays produced correct reactions for all staphylococci tested. Only the Dry Spot Staphytect Plus test correctly identified all 52 MRSA strains. For the other tests kits, sensitivity of MRSA detection was lower than for MSSA isolates. Depending upon the local MRSA prevalence and the parameter of interest (sensitivity or specificity), these test systems may be useful for routine diagnostic purposes.

Anti-Bacterial Agents↗
PubMed1981

Comparison of three immunological tests for seroepidemiological purposes in human echinococcosis.

The comparative sensitivity and specificity of three immunodiagnostic tests for hydatid disease--the latex agglutination (LA) test, the indirect haemagglutination (IHA) test and the counterimmunoelectrophoresis (CIEP) test--were studied. The results of the IHA test were evaluated using three positivity criteria. Antigen from the same source--hydatid cyst fluid pool of ovine origin--was utilized for the three tests. A total of 123 sera were studied; 30 preoperative sera from patients with other parasitic and infectious diseases, 30 sera from patients with non-parasitic diseases and 30 sera from normal blood donors. The results of the three tests were correlated to the immunoelectrophoresis (IEP) test based on the presence of arc 5. The same sensitivity (86.7%) and specificity (99%) was obtained in LA, CIEP and IHA tests when the minimum cross-reactivity titre criterion was employed for the latter. Non-specificity was relatively high (13%) in the IHA test when only serological reactivity was considered. The sensitivity of the IHA test decreased (83.3%) using the diagnostic titre criterion and non-specific reactions were eliminated. All three tests correlated well with the IEP test.

Antibodies↗
PubMed2006

A simple and practical agglutination assay for human leucocyte antigen-B27 typing.

BACKGROUND AND OBJECTIVES: The human leucocyte antigen (HLA) B27 is the most frequently typed single antigen that is associated with diseases. Here, we describe a simple and rapid particle agglutination assay (PaGIA) for HLA-B27 typing. MATERIALS AND METHODS: Superparamagnetic particles were coated with a monoclonal antibody to HLA-B27 and subsequently used for testing. Anticoagulated whole-blood samples were obtained from healthy blood donors (n = 194) with known HLA patterns and from patients (n = 51) who had been typed positive for HLA-B27 by flow cytometry. RESULTS: The particles agglutinated only after incubation with HLA-27-positive blood samples, using the ID-microtyping system. Positive reactions were clearly distinguishable from negative reactions in all samples tested. Flow cytometric HLA-B27 typing revealed an indeterminate result in one patient. CONCLUSIONS: The new HLA-B27 PaGIA is suitable for rapid typing of HLA-B27. The assay is simple and easy to perform, and can be implemented in any routine laboratory.

Agglutination Tests↗
PubMedDate not supplied

A study of weak subgroups of blood group A with an antiglobulin-latex test.

The distribution of the blood group A antigen on subgroups A1, A2, A3, Ax, and Ae1 was studied by means of a highly sensitive and specific antiglobulin-latex test. Marked variability in the cell to cell expression of A antigen was observed in A1 and A2 cells. The weaker subgroups (A3, Ax, and Ae1) were characterized by a minor population of cells containing A antigen and a major population with no detectable A sites. Comparison of the antiglobulin-latex test with the fluorescent antibody technique indicates that the former is approximately 10,000 times more sensitive in detecting cell bound antibody, and thereby membrane antigens. The results of this study demonstrate that within each of the A subgroups (with the possible exception of A1) there exist erythrocytes that apparently lack recognizable A antigen sites. The number of such cells is greatest in the weakest subgroups (Ax and Ae1).

ABO Blood-Group System↗
PubMed1993

Rapid detection and characterization of sialic acid-specific lectins of Helicobacter pylori.

A particle agglutination assay (PAA) using fetuin (Ft) covalently coupled to carboxylate-modified latex (CML) particles was evaluated for rapid detection of sialic acid-specific haemagglutinins/lectins (SALs) of Helicobacter pylori isolates which bind sialoglycoconjugates. Sixty-three percent (20/32) of the isolates examined gave a positive PAA test. Cell-bound SALs were extracted by washing the bacteria with deionized water or isotonic saline, and their expression was influenced by pH and culture conditions. The Ft-CML reactivity of the PAA-positive isolates was inhibited by bovine submaxillary mucin, transferrin, fetuin, orosomucoid, vitronectin and lactoferrin in a manner which suggested that the isolates contain a lectin recognizing the alpha(2-6) linkage of terminal sialic acid. Western blots of strain NCTC 11637 SALs probed with horseradish peroxidase (HRP)-labelled Ft identified three bands (MW 64 kD, 62 kD, 56 kD) which also reacted with HRP-labelled mucin, transferrin, lactoferrin, orosomucoid, vitronectin and laminin. Sera from patients with a H. pylori infection and one polyclonal rabbit antiserum (strain NCTC 11637) also reacted with the SALs. Immunogold labelling of a polyclonal rabbit antiserum raised against the 64 kD protein of strain NCTC 11637 that reacted strongly with Ft-CML showed that abundant SALs were loosely cell-associated with the cell surface of both spiral and coccoidal forms of H. pylori. SALs were also present in low amounts on the surface of strain NCTC 11638 and 66, a clinical isolate that did not react with Ft-CML.

Animals↗