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Relationships of total protein, specific gravity, viscosity, refractive index and latex agglutination to immunoglobulin G concentration in mare colostrum.

A colostrum sample was collected within 24 h after foaling from 27 mares and from 10 other mares a milk sample was collected several weeks post partum. Immunoglobulin G concentrations were determined quantitatively by radial immunodiffusion and semi-quantitatively using a commercial latex agglutination test. Total protein, specific gravity, viscosity and refractive index were determined and their relationships to the immunoglobulin G concentration analysed. All parameters correlated with the immunoglobulin G concentration. The latex agglutination test divided the colostrum samples into three groups with different means for immunoglobulin G and total protein concentrations. Specific gravity and the latex agglutination test were found to be the methods best suited for on-farm evaluation of colostrum quality.

Animals↗

Particle agglutination assays to identify fibronectin and collagen cell surface receptors and lectins in Aeromonas and Vibrio species.

A rapid particle agglutination assay (PAA) utilizing latex beads coated with connective tissue and serum proteins was evaluated for its ability to identify fibronectin, collagen (types I and IV), fibrinogen, and transferrin cell surface receptors on Vibrio and Aeromonas strains isolated from diseased fish, human infections, and the environment. Similar tests were performed to screen for cell surface lectins. Vibrio as well as Aeromonas strains were found to bind connective tissue proteins (collagen types I, II, and IV and fibronectin), serum proteins (i.e., fibrinogen), and glycoproteins (bovine submaxillary mucin, hog gastric mucin, orosomucoid, and fetuin) immobilized on the latex particles. The specificity of the agglutination reaction was studied by particle agglutination inhibition assays performed by preincubating bacterial suspensions in solutions containing either gelatin (for the various connective tissue protein PAA reagents) or sialic acid-rich glycoproteins (for the various glycoprotein PAA reagents). Expression of cell surface receptors for connective tissue proteins was found to depend on culture methods.

Aeromonas↗

Evaluation of a microtiter latex agglutination test for histoplasmosis.

Experiments were designed to evaluate a Microtiter latex agglutination (Micro-LA) test, as a serological aid in the diagnosis of histoplasmosis, and to compare this test with the conventional microtiter-complement fixation (CF) test for histoplasmosis. Sera tested were from cases of acute and chronic pulmonary and disseminated histoplasmosis, as well as from individuals not having histoplasmosis. Ninety-seven percent of the cases of acute pulmonary histoplasmosis had positive Micro-LA tests, whereas 91% had positive CF tests. Ninety-six percent of the patients having chronic pulmonary histoplasmosis showed positive Micro-LA tests and 91% had positive CF tests. In contrast, 64% of the cases of disseminated histoplasmosis had positive Micro-LA tests, whereas 82% had positive CF tests. None of these differences was statistically significant. Although there were no significant differences in complement fixing and agglutinating antibody cross-reactivity with Blastomyces antigens, more patients demonstrated CF titers than Micro-LA titers. Sera from patients with acute and chronic histoplasmosis showed higher Micro-LA titers than CF titers, whereas sera from cases of disseminated histoplasmosis showed higher CF titers. Histoplasmin skin testing has less of a boosting effect on agglutinating antibodies than on CF antibodies to histoplasmin. Anticomplementary sera can be used in the Micro-LA test. This test is simple to perform, and results can be obtained in 2 to 4 hr.

Acute Disease↗

Mycobacterial agglutination-inhibition test.

The mycobacterial agglutination-inhibition test (MAIT) is described as a modification of the mycobacterial agglutination of Schaefer (MATS). It is an organized assemblage of absorption and agglutination tests which is especially useful in the identification of Mycobacterium avium isolates normally excluded from serological identification because of unstable cell suspension characteristics. The identity of 100 unstable M. avium and 3 unstable Mycobacterium intracellulare isolates was determined by the MAIT method and by animal pathogenicity tests. The comparability of the MAIT and MATS methods was demonstrated by testing 10 stable M. avium and 10 stable M. intracellulare isolates by both methods and obtaining the same serological identification for each culture. The reproducibility of the MAIT was confirmed when the same result was obtained in three consecutive tests on each of the 123 cultures used in this evaluation.

Absorption↗

Evaluation of a latex agglutination kit (Virogen Rotatest) for detection of bovine rotavirus in fecal samples.

The performance of the Virogen Rotatest latex agglutination test (LAT) was evaluated for detection of bovine rotavirus antigen. Sixty-three fecal samples from diarrheic calves were collected from November 1999 to May 2000 and screened by LAT, the Rotazyme II enzyme-linked immunosorbent assay (ELISA), and virus isolation (VI) followed by an anti-rotavirus fluorescent-antibody (FA) test to detect the presence of group A rotavirus antigen. Of the 63 samples screened by VI-FA, 33 (58%) tested positive for rotavirus antigen. When the results from the LAT were compared to those from VI-FA, the "gold standard" for detection of bovine rotavirus in fecal samples, the sensitivity and specificity were found to be 87.8 and 73.3%, respectively. Latex agglutination compared with ELISA (the reference method) showed 100% sensitivity and 96.3% specificity, and when ELISA was compared with VI, the sensitivity was 84.8% and the specificity was 73.3%. Latex agglutination is easy to perform in a short time and does not require expensive equipment or skilled personnel, and the reagents have long shelf lives. These factors make the LAT suitable and highly efficient for use in a clinical laboratory as a rapid screening test for bovine rotavirus.

Animals↗

Agglutination of mouse erythrocytes by Eperythrozoon coccoides.

Erythrocytes from blood of mice infected with Eperythrozoon coccoides for 3 or 4 days agglutinated spontaneously. Washed E. coccoides particles agglutinated washed erythrocytes of uninfected mice. E. coccoides-mediated agglutination of normal mouse erythrocytes would be an excellent system for studies of bacterial adhesion.

Adhesiveness↗

Contribution of humoral and cellular factors to the resistance to experimental infection by Pseudomonas aeruginosa in mice. II. Opsonic, agglutinative, and protective capacities of immunoglobulin G anti-Pseudomonas antibodies.

The opsonic, agglutinative, and mouse protective capacities of anti-Pseudomonas antibodies in immune and normal human immunoglobulin G (IgG) preparations were investigated. Opsonic activities of the immune IgG preparations correlated well with their protective activities. Antibodies present in normal IgG showed a substantial agglutinative activity but were poorly protective. Anti-Pseudomonas antibodies of both normal and immune IgG preparations were directed against the same serotype antigen as demonstrated by absorption experiments. Immune anti-Pseudomonas IgG antibodies fixed complement very efficiently as demonstrated by opsonophagocytic and hemolytic tests. Natural anti-Pseudomonas IgG antibodies fixed complement very poorly although they promoted phagocytic killing of bacteria only in the presence of heat-labile serum factors. It was concluded that, although agglutination can be used for qualitative measurement of antibacterial antibodies, it fails to measure their functional capacities.

Animals↗

Serological grouping of meningococci and encapsulated Haemophilus influenzae strains by latex agglutination.

The latex agglutination method, utilizing antibody-coated latex particles, was adapted for serogrouping of Neisseria meningitidis and serotyping of encapsulated Haemophilus influenzae strains from agar plates. It was found to give more clear-cut results than conventional slide agglutination. A 100% agreement with the antiserum agar method was found for all strains isolated from blood or cerebrospinal fluid. Many meningococcal strains from nasopharyngeal carriers are autoagglutinable, but some of these gave a positive reaction with the group B latex reagent, although they were negative by the antiserum agar method. The latex agglutination method has several advantages over others: the lack of autoagglutination, easy performance, easy interpretation, and very low consumption of antisera.

Blood↗

Analysis of parameters affecting the hemagglutination activity of Escherichia coli possessing colonization factor antigens: improved medium for observing erythrocyte agglutination.

The hemagglutination (HA) activity of two strains of Escherichia coli, each possessing different colonization factor antigens (CFA), was examined under different test conditions. The effects of ionic strength, temperature, pH, cations, and reaction surface on erythrocyte (RBC) agglutination were analyzed. Strain H-10407 (CFA/I) caused the agglutination of human, bovine, and chicken RBC, whereas strain CL-9699 (CFA/II) agglutinated only bovine and chicken RBC. The HA activity of both strains increased with decreasing ionic strength, pH, and temperature, the effects of temperature being negligible at low ionic strength. When accounting for ionic strength, the presence of Ca2+, Mg2+, Fe2+, or Fe3+ ions did not increase the HA activity of these bacteria. Optimum conditions for HA of reactive RBC by bacteria included low ionic strength (less than 50 mM) and slightly acidic pH (6.0 to 7.0). Use of a low-ionic-strength medium permitted application of microtitration methods to visualize the HA reactions. Storage of RBC in low-ionic-strength medium did not change their HA properties, and the use of this medium proved superior to saline in overcoming HA variation observed with different preparations of RBC.

Animals↗

Evaluation of the latex slide agglutination test for identification of Staphylococcus aureus.

This study evaluated the reliability of the latex slide agglutination test for identifying Staphylococcus aureus. A total of 806 clinical isolates of staphylococci were tested for latex agglutination, clumping factor, and free coagulase. Positive latex tests occurred in 98.3% of coagulase-positive strains, whereas 99.6% of coagulase-negative strains gave negative latex tests. It is concluded that in most instances, the latex slide agglutination test is a reliable method for identifying S. aureus in the clinical laboratory.

Bacterial Proteins↗

Clinical studies of a new latex particle agglutination test for detection of Haemophilus influenzae type b polyribose phosphate antigen in serum, cerebrospinal fluid, and urine.

A new latex particle agglutination test for direct detection of Haemophilus influenzae type b polyribose phosphate antigen in serum, cerebrospinal fluid, or urine was evaluated from studies at four clinical centers. Although use of a serum buffer significantly reduced inconclusive agglutination of the latex particles, the retesting of serum samples, after heat inactivation and dilution, resolved all serum samples, with one exception, as reactive or nonreactive for the presence of the polyribose phosphate antigen. A clinical accuracy of 100% was obtained for the latex particle agglutination method in respect to its capability for detection of polyribose phosphate antigen in all patients with confirmed infection by H. influenzae type b.

Haemophilus Infections↗

Direct identification of Staphylococcus aureus in blood culture fluid with a commercial latex agglutination test.

A commercial latex agglutination slide test (SeroSTAT Staph, Scott Laboratories, Inc., Fiskeville, R.I.) accurately identified Staphylococcus aureus when applied directly to blood culture fluid containing staphylococci. This latex agglutination test exhibited 100% accuracy when 30 seeded aerobic and anaerobic radiometric blood cultures (15 strains of S. aureus, 15 strains of other staphylococcal species) were tested blindly. In 36 actual clinical specimens yielding 16 isolates of S. aureus and 20 isolates of Staphylococcus epidermidis, 94.4% accuracy was achieved. The latex agglutination test provided positive test results before objective criteria of blood culture positivity such as radiometric growth indices and Gram stains became positive.

Aerobiosis↗

Evaluation of a latex agglutination test for detection of antibodies to rubella virus in selected sera.

A new latex agglutination test for rubella virus was used to test sera with a hemagglutination inhibition titer of less than or equal to 8 (97 specimens) or greater than or equal to 256 (158 specimens). Maximum latex agglutination test sensitivity was achieved when low-titer sera were tested undiluted and high-titer sera were diluted (1:10). A modification of the protocol of the manufacturer resulted in a latex agglutination-hemagglutination inhibition agreement of 96.7%.

Antibodies, Viral↗

Detection of bacterial antigens in body fluids with the Wellcogen Haemophilus influenzae b, Streptococcus pneumoniae, and Neisseria meningitidis (ACYW135) latex agglutination tests.

The Wellcogen Haemophilus influenzae b, Streptococcus pneumoniae, and Neisseria meningitidis (ACYW135) latex agglutination tests (Wellcome Diagnostics, Dartford, England) were evaluated as methods to detect bacterial antigens in cerebrospinal fluid (CSF), urine, and serum from patients with meningitis or sepsis. Antigen was detected in 92% of CSFs from H. influenzae b, 100% of CSFs from N. meningitidis groups A and Y, 36% of CSFs from N. meningitidis group C, and 69% of CSFs from pneumococcal meningitidis patients. Serum samples presented a problem, with a few false-positive or possible cross-reactions. The Wellcogen latex agglutination tests were more sensitive than the Bactogen (H. influenzae type b) latex agglutination test and the Phadebact (S. pneumoniae) coagglutination test.

Antigens, Bacterial↗

Evaluation of a C-reactive protein latex agglutination detection test with sera from patients with sexually transmitted diseases.

A total of 149 sera, including 79 pre- and posttreatment sera from 33 patients with disseminated gonococcal infections, 18 from patients with uncomplicated gonococcal infections, 6 from patients with pelvic inflammatory disease, 4 from patients with genital Chlamydia trachomatis infections, and 42 from normal volunteers, were examined for C-reactive protein with a latex agglutination C-reactive protein detection kit (Difco Laboratories, Detroit, Mich.). Results were quantitated with LC-Partigen C-reactive protein radial immuno-diffusion plates (Calbiochem-Behring, La Jolla, Calif.). Positive latex agglutination results were observed in all of the pretreatment sera and some of the posttreatment sera of patients with disseminated gonococcal infections and in two sera from patients with pelvic inflammatory disease, which corresponded to quantitative C-reactive protein levels in the radial immunodiffusion plates. C-reactive protein levels were not detectable in the serum samples from normal volunteers or patients with uncomplicated gonococcal infections or genital chlamydial infections. Positive latex agglutination occurred as early as 20 s in sera with high C-reactive protein levels, and all positive results were observed within 90 s of the 3-min test limit. Positive latex test results were obtained with C-reactive protein levels as low as 1 mg/dl (10 micrograms/ml).

C-Reactive Protein↗

Receptor-specific agglutination tests for detection of bacteria that bind globoseries glycolipids.

Specific binding to the globoseries of glycolipid receptors explains the adherence of uropathogenic Escherichia coli to host cells. The minimal receptor disaccharide Gal alpha 1----4Gal beta [galactose alpha (1----4)galactose beta] is recognized by most attaching clinical isolates. However, wild-type isolates can express adhesins with several different receptor specificities. Bioassays do not permit separate analysis of each receptor specificity, since the target cells contain multiple potentially receptor-active molecules. In this study, bacterial adhesins were analyzed by using receptors immobilized into latex beads in one of two ways. In one way, di- and trisaccharides were covalently linked via a spacer arm to latex beads coupled with bovine serum albumin. In the other way, receptor-active glycolipids were coated onto the bovine serum albumin-latex beads. The latex beads were subsequently used for agglutination by using type strains with known receptor specificity. The composition was optimized regarding receptor structure and size of latex beads. Gal alpha 1----4Gal beta was as active as the trisaccharide derivative Gal alpha 1----4Gal beta 1----3glucose or Gal alpha 1----4Gal beta 1----3glucosamine. Among the natural glycolipids tested, globotetraosylceramide was the most active. Subsequently, the sensitivity and specificity of the Gal alpha 1----4Gal beta-latex and globotetraosylceramide-latex reagents were compared for 733 E. coli urinary isolates. Hemagglutination of human erythrocytes was used as the positive standard. No significant difference in the specificity or sensitivity of the latex reagents was found; the sensitivity ranged from 86%, when isolates agglutinating human erythrocytes of blood groups P1 and p were included, to 93%, when those isolates agglutinating erythrocytes of blood group p were excluded. These reagents provide tools for bacterial identification in patients with urinary tract infection.

Adhesins, Escherichia coli↗

Commercial latex agglutination test for detection of Clostridium difficile-associated diarrhea.

A commercially available latex agglutination test for Clostridium difficile was compared with a cell culture cytotoxin assay and bacteriological culture for the laboratory diagnosis of C. difficile-associated diarrhea and colitis (CAD). Stool specimens from 626 patients were tested by the three methods, and specimens from 118 patients (19%) were positive by at least one of the methods. The results of the three tests agreed in 88% of the specimens tested, overall, but they agreed in only 34% of the 118 positive specimens. Ninety-three patients were evaluated to assess the significance of positive and negative results for each assay. Of 40 patients found to have CAD, 70% were positive by the cytotoxin assay, 78% were positive by the latex agglutination test, and 90% were culture positive. Of 53 patients who did not have CAD, 2% were positive by the cytotoxin assay, 8% were positive by the latex test, and 4% were culture positive. The detection of CAD was improved by using the tests in combination, and 97% of specimens positive by two or three methods were from patients who had CAD. Testing of multiple specimens from individual patients also increased the sensitivity of detection of CAD. The results suggest that the latex agglutination test may be useful for rapid diagnosis of CAD, especially in laboratories that lack cell culture facilities. However, the accuracy of CAD detection is improved when the latex test is used in combination with culture or the cytotoxin assay.

Adult↗

Detection of antibodies to human immunodeficiency virus by latex agglutination with recombinant antigen.

Recombinant human immunodeficiency virus (HIV) env antigen was attached to polystyrene particles, and these complexes were used to develop the first latex agglutination assay for antibodies to HIV. A total of 95 positive and 116 negative human serum samples were assayed for antibodies to HIV by latex agglutination, and results were compared with those of a commercial enzyme immunoassay. Latex agglutination was also compared with, and found to be completely concordant with, Western blot (immunoblot) analysis with virion antigens.

Antibodies, Viral↗