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PubMed2004

Simple, rapid latex agglutination test for serotyping of pneumococci (Pneumotest-Latex).

The "gold standard" for epidemiological typing of Streptococcus pneumoniae (pneumococcus) is the capsular reaction test (Neufeld test) with antisera against the 90 pneumococcal polysaccharide capsules, i.e., serotyping. The method is labor intensive and requires a certain level of experience to be performed satisfactory, and thus it has been restricted for use in specialized reference or research laboratories. Surveillance of the serotype distribution of pneumococci that cause infections is important to secure an optimal composition of pneumococcal vaccines and to monitor antibiotic resistance in pneumococci. At Statens Serum Institut, a simple latex agglutination test for serotyping of pneumococci has been developed. The Pneumotest-Latex kit consists of 14 different pooled pneumococcus antisera (pools A to I and pools P to T) applied to latex particles. In a blind test of 352 isolates (with all 90 serotypes represented), 336 (95.5%) were typed or grouped correctly by the Pneumotest-Latex; in addition, 2 (7%) of 30 strains regarded as nontypeable or rough strains were serotyped, and the serotypes of these two isolates were confirmed by the capsular reaction test with type-specific antisera. The Pneumotest-Latex seems to be a sensitive method for serotyping or grouping of the majority of pneumococcal strains. By use of this ready-to-perform latex agglutination kit (Pneumotest-Latex), serotyping of pneumococci can gain more ground as a tool in prevention of pneumococcal diseases.

Bacterial Typing Techniques↗
PubMed2004

Evaluation of the methicillin-resistant Staphylococcus aureus (MRSA)-Screen latex agglutination test for detection of MRSA of animal origin.

Methicillin (oxacillin)-resistant staphylococci (MRS) have emerged as major clinical and epidemiological pathogens, and there have been frequent reports of MRS infections in the veterinary field. The MRSA-Screen latex agglutination test (Denka Seiken Co., Ltd., Tokyo, Japan) was compared with an oxacillin agar screen test, MIC determination, and mecA PCR assay, the "gold standard." In an analysis of 15 mecA-positive and 48 mecA-negative S. aureus animal isolates, as well as 9 mecA-positive and 147 mecA-negative, coagulase-negative staphylococcal animal isolates, the latex agglutination test surpassed the widely used oxacillin agar screen method and MIC determination, with a sensitivity and a specificity of 100%. The MRSA-Screen test is a reliable and rapid method of detecting MRS in the veterinary field.

Animals↗
PubMed1978

Rapid grouping of beta-hemolytic streptococci by latex agglutination.

Latex agglutination was compared with fluorescent-antibody staining with group A conjugate and Lancefield precipitation for grouping of beta-hemolytic streptococci. Latex agglutination correctly grouped 98.8% of 82 group A streptococci and more than 95% of 187 group B, C, or G streptococci. Occasional cross-reactions occurred between groups A and C and groups B and G.

Fluorescent Antibody Technique↗
PubMed1978

Broadly reacting precipitating and agglutinating antigen of leptospirae.

A saprophytic Leptospira biflexa strain of equine origin was found which cross-reacts with immune rabbit antisera to 14 pathogenic Leptospira interrogans serotypes. Sera from goats experimentally inoculated with the saprophyte showed multiple low-level cross-agglutination reactions against a battery of live L. interrogans serotypes. Sonically treated and saline-extracted suspensions of the L. biflexa strain and serotypes canicola, icterohaemorrhagiae, and pomona yielded a common precipitating protein antigen that was detected by immunodiffusion and immunoelectrophoresis with all of the antileptospiral sera examined. In cross-absorption and gel diffusion tests, the precipitinogen from each of the strains was shown to be identical. Formaldehyde treatments and heating at 100 degree C suggest that the cellular location of the common antigen is either somatic or subsurface, and Sephadex G-200 gel filtration enabled the isolation of the active fraction of the L. biflexa antigen. Monoprecipitin sera against the common antigen of L. biflexa were produced by immunizing rabbits with specific precipitates in agar. In gel diffusion and immunoelectrophoresis tests the antisera with each of the soluble antigens developed a single precipitin formation, and the antisera agglutinated formolized and heated whole-cell suspensions of serotypes canicola, icterohaemorrhagiae, and pomona at low dilutions. The soluble L. biflexa antigen was evaluated as an immunogen and in passive immunity tests for protection against death and kidney infection in hamsters. No cross-protection occurred when the hamsters were challenged with virulent leptospires. In contrast, the animals vaccinated or administered hamster immune serum before challenge died earlier than the control animals.

Animals↗
PubMed2001

Unique monoclonal antibody recognizing the third extracellular loop of CXCR4 induces lymphocyte agglutination and enhances human immunodeficiency virus type 1-mediated syncytium formation and productive infection.

To increase insight into the structural basis of CXCR4 utilization in human immunodeficiency virus type 1 (HIV-1) infection, a new generation of three monoclonal antibodies (MAbs) was developed in WKA rats. The A80 MAb, which binds an epitope in the third extracellular loop (ECL3) of CXCR4, has unique biologic properties that provide novel insights into CXCR4 function. This agent enhanced syncytium formation in activated human peripheral blood mononuclear cells (PBMC) infected with X4 or R5 and CEM cells infected with X4 HIV-1 strains. Exposure to A80 increased the productive infection of activated CD4(+) T cells and CEM cells with R5 and X4 viruses, respectively. This antibody uniquely induced agglutination of PBMC and CEM cells but did not activate calcium mobilization. Agglutination induced by A80 was inhibited by stromal cell-derived factor 1, T22, and phorbol 12-myristate 13-acetate but was not significantly altered by pretreatment of cells with pertussis toxin, wortmannin, or MAbs to LFA-1, ICAM-1, ICAM-2, and ICAM-3. The binding of the A145 and A120 MAbs was mapped to the N-terminal extracellular domain and a conformational epitope involving ECL1 and ECL2, respectively. Both of these MAbs inhibited HIV-1 infection and lacked the novel properties of A80. These results suggest a new role for CXCR4 in homologous lymphocyte adhesion that is ligand independent and in HIV-1 infection.

Amino Acid Sequence↗
PubMed1984

Sequential study of C reactive protein in neonatal septicaemia using a latex agglutination test.

The usefulness of serial study of C reactive protein in the early detection of neonatal septicaemia was evaluated in a neonatal unit using a commercially available latex agglutination slide test as a rapid screening method and electroimmunoassay as a reference method for C reactive protein determination. A positive latex test was obtained in 11 infants with verified septicaemia (positive blood culture), two infants with clinically evident infection but without bacteriological confirmation, one infant with recurrent chest infection due to Pseudomonas aeruginosa, and one infant who showed signs of birth asphyxia with meconium aspiration, but was not infected. Positive latex test results correlated with raised concentrations of C reactive protein, measured by immunoassay. In some instances, however, concentrations of C reactive protein in excess of 12 mg/100 ml gave weaker agglutination results in the slide test, which could be interpreted as negative results. In a sequential study of the infected infants, 6.3% of the values recorded on a slide test were false negatives. In contrast, false positive values were observed on a slide test in 1.9% of 27 non-infected infants. The higher percentage of false negative values may be due to the presence of excess antigen in the sera of some infected children. It is suggested that the latex test should be carried out on suitable dilutions of serum. Although the slide test may reliably indicate infection at an early stage in neonates, the C reactive protein response is non-specific, as seen in a non-infected infant who showed signs of birth asphyxia with meconium aspiration. Provided the non-specific nature of the C reactive protein response is recognised, the latex test may be a useful serum measurement for early diagnosis of neonatal septicaemia of the newborn. The test has the advantage of being performed easily, quickly, and cheaply.

C-Reactive Protein↗
PubMed1985

Detection of rotavirus by a latex agglutination test, Rotalex: comparison with electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay.

A commercially available latex agglutination test, Rotalex (Orion Diagnostics, Finland), for detecting rotaviruses was evaluated in comparison with four other tests (electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay) routinely used in our laboratories. Although Rotalex was the least complex method, it showed lack of specificity and sensitivity when carried out according to the manufacturer's instructions. Four basic modifications of Rotalex are described. These include the use of Hank's balanced salt solution, increasing the incubation time to 20 min, reading the agglutination result by an experienced observer, and the use of 50 mm square glass plates. The modified procedure gave results which were comparable with those obtained by electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay. The latter techniques, when used to detect rotavirus, all gave similar results.

Electrophoresis, Polyacrylamide Gel↗
PubMed1989

Discrepant toxoplasma latex agglutination test results.

The analysis of 4450 toxoplasma serology results showed that 59 (1.3%) latex agglutination reactions were not confirmed in the dye test. These discrepant results were associated with an unspecified IgM antibody but not associated with kit batch variation, inactivation of sera, concurrent cytomegalovirus infection, or the presence of hepatitis B virus "e" antigen. The latex agglutination test is useful as a screen for toxoplasma infection but false positive reactions do occur. Patients at risk of severe toxoplasmosis should be investigated by additional tests.

Animals↗
PubMed1995

Latex agglutination for rapid detection of Pseudomonas pseudomallei antigen in urine of patients with melioidosis.

A latex agglutination test for the detection of Pseudomonas pseudomallei antigen in urine was evaluated for the rapid diagnosis of melioidosis. With unconcentrated urine, antigen was detected in only 18% of patients with melioidosis overall. However, when urine was concentrated 100-fold, antigen was detected in 47% overall and in 67% of patients with septicaemia or disseminated infection, in whom a rapid diagnosis is most important. The specificity of the test was 100%. These results compared favourably with an enzyme immunoassay. This latex agglutination test is a simple, rapid and highly specific method of diagnosing melioidosis, and will be particularly useful in areas with limited laboratory facilities.

Antigens, Bacterial↗
PubMed1995

Clinical evaluation and reproducibility of the Pastorex Aspergillus antigen latex agglutination test for diagnosing invasive aspergillosis.

AIMS: The performance of the Pastorex Aspergillus antigen latex agglutination test for the detection of galactomannan in sera of patients at risk for invasive aspergillosis was evaluated, and the impact of storage on the reproducibility of the antigen titre was tested. METHODS: During a one year period, 392 serum samples were obtained from 46 patients at risk for invasive aspergillosis and tested for the presence of galactomannan using an Aspergillus latex agglutination test (Pastorex). Twenty three positive serum samples which had been stored at -20 degrees C for 2-16 months were retrospectively retested. Furthermore, two positive serum samples were stored at -20 degrees C and -70 degrees C and prospectively tested at three month intervals for a period of 15 months. RESULTS: The Pastorex Aspergillus test was positive in eight patients with microbiological, radiological, or histological evidence for invasive aspergillosis, but was negative in the initial serum sample from five of these patients. In two patients with histological evidence for invasive aspergillosis no positive reaction was found in six samples. Six of 13 (45%) serum samples which had been stored at -20 degrees C for longer than six months had lost reactivity, while one of 10 (10%) samples had lost reactivity when stored up to six months. Two serum samples which had been stored at -20 degrees C and -70 degrees C and prospectively retested at three month intervals for 15 months, maintained stable antigen titres. CONCLUSIONS: The Pastorex Aspergillus test is too insensitive to diagnose invasive aspergillosis in an early stage, but may contribute to the diagnosis when cultures remain negative and serial samples are obtained. To maintain a good reproducibility, serum samples should be stored at -70 degrees C when the period of storage exceeds six months.

Adolescent↗
PubMed1999

A latex slide agglutination test for rapid detection of antimyeloperoxidase antibody.

AIM: To develop and test a new latex slide agglutination test (MPO-LSAT) to detect antimyeloperoxidase (anti-MPO) antibody in serum. METHODS: Latex bead coating was adjusted to give maximum sensitivity by attending to latex size, MPO to latex ratio for coupling, ratio of diluted serum to MPO-latex, reaction time and temperature for coupling, and reaction time for agglutination. Inhibition studies were performed using MPO, proteinase 3, bactericidal/permeability increasing protein, and lactoferrin. RESULTS: There was very good correlation between this test and the conventional anti-MPO enzyme linked immunosorbent assay (ELISA): 81% of sera positive in the ELISA were positive by MPO-LSAT. MPO-LSAT results correlated better with IgM anti-MPO than with IgG anti-MPO. CONCLUSIONS: MPO-LSAT is a simple diagnostic test that is potentially useful in the clinical laboratory as a rapid screening tool for vasculitic diseases.

Antibodies, Antineutrophil Cytoplasmic↗
PubMed1985

Uncertainty in identification of blood group A subtypes by agglutination test.

Identification of the blood group A subtypes, i.e. A1, A2, and A1-A2 intermediate (Aint), by agglutination test, particularly in AB red cells, is ambiguous. The expressions of A subtypes in red blood cells are the consequences of diverse formations of the A substances by the action of three types of blood group N-acetyl-galactosaminyl-transferases controlled by A1, A2, and Aint genes. Therefore, the A subtypes are more directly identified by examining the kinetic characters of A-enzymes existing in plasma. Several Black AB subjects classified as non-A1 by the agglutination test were identified as A1B and AintB on the enzyme basis. A subject serologically classified as A1 had A2-enzyme in her plasma, i.e. she is genetically A2O or A2A2. The present and previous studies indicate that red cell A2 status is occasionally expressed as a result of the combination of Aint and B, and of A1 and superactive B. The imbalance between A1/A2 and A1B/A2B observed in some Black populations could be attributed to high frequencies of the Aint and B. sup. genes in Blacks.

ABO Blood-Group System↗
PubMed1985

Cell surface alterations in murine leukaemia P388 adriamycin-resistant cells: studies on lectin-induced agglutination and rearrangement of lectin receptors.

Cell surface alteration was studied in a subline of murine lymphocytic leukaemia resistant to the broad-spectrum anticancer agent adriamycin (P388/ADR) employing concanavalin A(Con A)-induced agglutination and rearrangement of lectin receptors. Con A induced more agglutination of P388/ADR when compared to the drug-sensitive parental cell line (P388/S). Studies on the redistribution of Con A and Ricinus communis agglutinin-I revealed a high percentage of P388/ADR showing internalized fluorescence, while a majority of P388/S displayed a uniform distribution of fluorescence on the cell surface.

Animals↗
PubMed1981

Levamisole-induced neutropenia and agranulocytosis: association with HLA B27 leukocyte agglutinating and lymphocytotoxic antibodies.

Among 37 patients treated with levamisole for rheumatoid arthritis (n = 19), for Reiter's disease (n = 4) and for chronic articular brucellosis (n = 14) followed up during 6-12 months, 3 developed agranulocytosis and 3 severe neutropenia. Serum samples drawn before and during treatment were tested for leukocyte agglutinating and lymphocytotoxic antibodies. Leukocyte agglutinating antibodies were induced in 8 patients, in 5 of them in association with agranulocytosis or neutropenia. In 1 patient with agranulocytosis and in another one with neutropenia lymphocytotoxic antibodies were also induced. Two agranulocytotic and one neutropenic patient possessed HLA B27 antigen. In altogether 11 HLA B27 carriers the number of circulating neutrophils were significantly reduced during levamisole treatment when compared with those of patients lacking HLA B27 antigen.

Adult↗
PubMed1989

Comparison of two latex agglutination test kits for serum myoglobin in the exclusion of acute myocardial infarction.

The Myolex (Orion) and the RapiTex (Behringwerke) latex agglutination tests for the rapid detection of elevated levels of serum myoglobin were studied prospectively in patients suspected of acute myocardial infarction, who were admitted to hospital within 8 h of pain onset. Using admission blood samples drawn 3.4 +/- 2.0 h (mean +/- SD) after onset of symptoms, the negative predictive values of both tests were too low to use these assays in the early exclusion of myocardial infarction in the emergency department. However, the negative predictive values obtained with the second blood samples, drawn 4 h later, indicated that the myoglobin agglutination test could be of value in the exclusion of myocardial infarction.

Adult↗
PubMed2000

Evaluation of solid-phase chemiluminescent enzyme immunoassay, enzyme-linked immunosorbent assay, and latex agglutination tests for screening toxoplasma IgG in samples obtained from cats and pigs.

Serum samples from cats and pigs were analyzed by the solid-phase chemiluminescent enzyme immunoassay (SPCEI), enzyme-linked immunosorbent assay (ELISA), and indirect latex agglutination (ILA) methods. The SPCEI and ILA methods accurately analyzed Toxoplasma IgG (T-IgG) in both clinical and spiked samples from pigs and cats. The ELISA method accurately analyzed T-IgG in spiked samples from cats and pigs or clinical samples from pigs, but it did not accurately analyze T-IgG in clinical samples from cats. The antibody used in the ELISA kit did not cross-react with cat T-IgG. The SPCEI method that uses a stand-alone automated analyzer provided quantitative analysis, whereas the ELISA and ILA methods provided qualitative or, at best, semiquantitative analysis of T-IgG. The SPCEI and ELISA methods were rapid (60-90 minutes for 30 samples), whereas the ILA method required 13-15 hours for 30 samples. Although the three methods accurately distinguished positive from negative samples, the ILA method yielded many weakly positive results that were not confirmed by either the ELISA or SPCEI method. Thus, the indirect agglutination tests may give nonspecific responses at lower T-IgG concentrations.

Animals↗
PubMed1999

Evaluation of quantitative latex agglutination for detection of Cryptosporidium parvum, E. coli K99, and rotavirus in calf feces.

A new methodology for detection of rotavirus, Escherichia coli K99, and Cryptosporidium parvum in bovine fecal samples was developed based on a quantitative latex agglutination technique (QLAT). Calibrated microspheres coated with specific antibodies to 1 of the enteric pathogens are quantitatively agglutinated by the antigens present in diluted fecal sample. The test is performed in a 96-well flat-bottom plate. The samples were tested with a commercial enzyme-linked immunosorbent assay kit prior to being analyzed by QLAT. The calculated sensitivity and specificity are adequate for field conditions, because the amount of the pathogenic agents is generally high. The overall time to perform the test was about 20 minutes.

Animals↗
PubMed1988

Early diagnosis of acute myocardial infarction by myoglobin latex agglutination test.

Early elevation of the serum myoglobin level in acute myocardial infarction (AMI) has been noted for years. In this study, 39 patients admitted to the Coronary Care Unit with acute chest pain within 72 hours (mean 12 +/- 15 hours) or electrocardiographic changes suspected of acute myocardial infarction had a serum myoglobin latex agglutination test to evaluate its diagnostic accuracy in acute myocardial infarction. Of these 39 patients, 24 had documented acute myocardial infarction as their final diagnosis. By the time of admission, 18 of the 24 cases with infarction had positive myoglobin tests (sensitivity 75%). Of those 15 cases without myocardial infarction, 13 had negative myoglobin tests (specificity 87%). If only those 17 cases admitted within 5 hours of the onset of chest pain were analyzed, the serum myoglobin test became positive in 8 of 10 cases with documented AMI but the 2 cases with negative results turned positive 2 hours later (sensitivity 80% to 100%). Due to the fact that myoglobin tests were negative in all other 7 cases without infarction, the specificity was 7/7 (100%). In contrast, the creatine phosphokinase isoenzyme study was positive only in 3 of these 10 patients with documented AMI in the first blood sample taken during admission. In conclusion, the serum myoglobin latex agglutination test is a quick and reliable method that helps in the early diagnosis of acute myocardial infarction.

Adult↗