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Diagnosis of Mycoplasma pneumoniae infection by microparticle agglutination and antibody-capture enzyme-immunoassay.

The performance of two new commercial assays for the serological diagnosis of Mycoplasma pneumoniae infection (microparticle agglutination and antibody-capture enzyme-immunoassay) was studied using a panel of 169 serum samples from patients with Mycoplasma pneumoniae pneumonia and a control group. Both assays were shown to be sensitive and specific for diagnosis. The performance of the capture immunoassay, however, decreased in older patients, probably due to its inability to detect cases of reinfection without IgM antibody response.

Adolescent↗

Specificity of new peptide immunoassays versus other immunoassays and agglutination tests for detection of HIV antibody in African sera.

The specificity of three HIV EIAs using synthetic peptides was evaluated and compared with that of four commercial EIAs using recombinant proteins or whole virus lysates of HIV-1 or of HIV-1 and HIV-2, and of two tests based on agglutination of sensitized particles. A total of 734 West African sera were screened. The best results were obtained with tests using recombinant proteins. HIV tests using synthetic peptides able to distinguish between HIV-1 and HIV-2 antibodies showed comparable specificity, but some false negative results represent a major handicap of these peptide EIAs in screening for HIV antibodies.

Agglutination Tests↗

A new method for detection of anti-zona activity in human sera using latex agglutination reaction.

A slide latex test for detection of anti-zona pellucida activity in human sera was developed using a latex agglutination reaction. The latex reagent, made of polystyrene particles coated with solubilizing zona antigen(s), was found to give results comparable in sensitivity as well as specificity to those of the indirect immunofluorescence method as tested with anti-pig-zona antiserum and with human sera. Thus, the slide latex test was judged to be adequate for use instead of the indirect immunofluorescence technique.

Adolescent↗

Rapid detection of Haemophilus influenzae type b in cerebrospinal fluid by commercial coagglutination and latex agglutination kits.

The ability of two commercial kits to detect Haemophilus influenzae type b antigen in cerebrospinal fluid was evaluated. Results obtained by Bactogen, a latex agglutination test, and Phadebact, a coagglutination test, were compared to counterimmunoelectrophoresis, Gram stain and culture results. One hundred and seven specimens of cerebrospinal fluid were tested. Thirty were found to contain bacteria, 20 of which were Haemophilus influenzae type b. All 20 were positive by Bactogen and Phadebact testing, 19 were culture positive, 18 were positive by counterimmunoelectrophoresis and 15 had gram-negative bacilli seen on Gram stain. The culture negative specimen contained microscopically visible gram-negative bacilli and was from a patient on antimicrobial therapy who was previously Haemophilus influenzae type b culture positive. No false positives from other genera or the 77 culture negative specimens occurred with Phadebact, Bactogen or counterimmunoelectrophoresis.

Agglutination Tests↗

[Application of the modified mixed cell agglutination reaction (MCAR according to Davidsohn) in forensic serology; MCAR on adhesive tape (author's transl)].

In view of serological sensitivity, specificity and simple procedure of reaction pattern the modified mixed cell agglutination reaction (Davidsohn's MCAR) proved to be a very useful test for typing ABO isoantigens in forensic medicine. Its sensitivity surpasses in some cases, where very small amounts of isoantigens have to be detected, that of the absorption-elution reaction several thousand times. The capacity to demonstrate the localization of antigens seems to be more reliable than the original test method. The modified Davidsohn's MCAR (MCAR on adhesive tape) is expecially recommended when isoantigens which are localized on the surface of microscopic objects including fingerprints, dust, hairs, and histological sections, have to be determined. The usefulness of our modification in forensic medicine and the conditions which are essential for the application, are discussed.

Agglutination Tests↗

Group B streptococcal polysaccharide detection in the urine of neonates by staphylococcal co-agglutination.

Staphylococcal co-agglutination reagents were prepared by the sensitization of commercially available staphylococci with commercially available grouping serum. The reagents were able to detect from 31 ng/ml to 62.5 ng/ml of purified group B polysaccharide lacking type specificity, when 20 microliters of polysaccharide solution was used. The reagents were able to identify group B streptococcal polysaccharide in the concentrated urine of all six patients with positive group B streptococcal blood cultures (sensitivity = 100%). Two of 72 patients with negative blood cultures had positive tests when concentrated urine was tested (specificity = 97%). Prepared reagents had a sensitivity and specificity similar to that reported by others for commercially available reagents for the detection of group B polysaccharide at five to ten percent of the total cost.

Agglutination Tests↗

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↗

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↗

[A method for treatment of partially haemolysed blood with lost agglutination properties, allowing its blood group determination (author's transl)].

A method for treatment of partially haemolysed blood with lost agglutination properties is described, which allows a direct determination of its blood group. The method is based on the phenomenon of reversible agglomeration of erythrocytes after treatment of the blood with nonelectrolytic and electrolytic solutions. This phenomenon is well known in modern transfusion haematology (Huggins C. E., 1963). This method has been tested on 156 blood samples with anti-A, B and AB serums and on 47 blood samples with anti-A, B, AB and H serums, as well, with very good results.

Agglutination↗

Immunoglobulin free light chain assay using latex agglutination.

Under normal biological conditions, immunoglobulin light chains are generally formed in conjunction with heavy chains; however, small quantities of free light chains are also produced. Thus, two types of immunoglobulin light chains may be identified; immunoglobulin-bound light chains and free light chains. In mature neoplastic B cells, immunoglobulin synthesis, including that of the free light chain, is generally increased. Furthermore, elevated free light chain levels also occur in the presence of renal glomerular and tubular impairment. Thus, the quantitative and qualitative measurement of free light chains is of considerable clinical benefit. Although methods for measurement of free light chains do exist they are technically complicated and time consuming. Previously, we have reported a quantitative nephelometric immunoassay for measuring free light chain levels. We report here a refinement of this method using latex agglutination techniques. This new method enables quantitative measurement of free light chains in serum at levels as low as 1 mg/l and may allow the clinical application of serum free light chain estimation.

Adult↗

Prevention of cell agglutination and competence in a genetically transformable strain of Pneumococcus by D-glucosamine and D-galactosamine.

In the presence of amino sugars D-glucosamine and D-galactosamine no spontaneous competence could be observed in the highly transformable R6bd strain of Pneumococcus or it was decreased by several orders of magnitude. The highest inhibition of competence was detected when the amino sugar at a concentration 5 mg/ml of the medium was added not only to the transformation but also to the pretransformation medium. After a 150 min growth in the transformation medium in the presence of the amino sugar a 3--4-fold greater number of cells (as a viable count) could be detected as compared with the control without the amino sugar. It was found microscopically that the amino sugar prevents natural agglutination, which normally occurs in the competent culture. The role of specific amino sugar determinants for binding of the competence factor on the cell surface and the resulting inhibitory effect of these sugars on the development of competence are discussed.

Agglutination↗

Comparison of gelatin particle agglutination and hemagglutination inhibition tests for measles seroepidemiology studies.

The prevalence of measles antibody in Japan was surveyed with a newly developed gelatin particle agglutination (PA) test, and the results compared with those of the hemagglutination inhibition (HI) test. The two age-distribution curves of the PA antibody-positive rates at > or = 1:8 and > or = 1:32 were almost the same in all the age groups, except the less-than-1-year-old group for which the rate at > or = 1:8 was higher than that at > or = 1:32 (p < 0.05, chi 2 test). In the vaccinated children, all groups older-than-1-year of age had antibody-positive levels of 96% or more. In contrast, in the unvaccinated children, there was a sharp increase in antibody-positive rates between the 1- and 4-year-old groups, indicative that about 80% of the children were infected by wild measles virus at these ages. A significant number of PA antibody-positive specimens were antibody-negative (< 1:8) by HI. The percentage of specimens in this category, PA (+) but HI (-), was greatest in infants less than one year old, and least in young children, but it increased with age to 97% of the HI (-) specimens from adults of more than 20 years of age. The PA test therefore detected some measles antibodies that HI could not. This test is simple and useful for making serosurveys in both developed and developing countries.

Adolescent↗

Immunoglobulin concentration in serum and seminal fluid from men with and without sperm-agglutinating antibodies.

Serum and seminal fluid from infertile men with head-to-head (H-H) or tail-to-tail (T-T) sperm-agglutinating antibodies and fertile men devoid of sperm antibodies were examined for immunoglobulin (Ig)G, IgA, IgM, IgD, and IgE. Only IgG, IgA, and IgE were demonstrated in the seminal fluid. No statistically significant differences in the serum or seminal fluid immunoglobulin concentrations were found between the fertile and the two infertile groups of men examined.

Antibodies↗

A new rapid slide agglutination test for amniotic fluid phosphatidylglycerol: laboratory and clinical correlation.

A new rapid slide agglutination test (Amniostat-FLM/AFLM) for amniotic fluid phosphatidylglycerol has been developed. One hundred ninety-three samples of amniotic fluid were tested for lecithin/sphingomyelin (L/S) ratio and phosphatidylglycerol by thin-layer chromatography and fluorescence polarization and compared with the AFLM. All four tests agreed in 123/193 cases. Phosphatidylglycerol and AFLM were concordant in 176/193 cases. From these samples, data were available on 105 neonates, of whom 74 were delivered within 72 hours of amniocentesis. All positive AFLM tests were associated with absence of hyaline membrane disease, of 22 negative tests, eight babies had hyaline membrane disease. Thus, preliminarily, it appears that this is a useful, rapid test for assessing fetal lung maturity, which like other tests is reliable when indicating maturity but has a high incidence of falsely immature results.

Agglutination Tests↗

Laboratory and clinical evaluation of a rapid slide agglutination test for phosphatidylglycerol.

Amniostat-FLM is a recently developed rapid slide agglutination test for amniotic fluid phosphatidylglycerol. We compared phosphatidylglycerol quantitated by two-dimensional thin-layer chromatography with phosphatidylglycerol determined by Amniostat-FLM in 180 amniotic fluid samples. If any level of phosphatidylglycerol by thin-layer chromatography, including a trace, was considered positive, then phosphatidylglycerol by thin-layer chromatography and Amniostat-FLM were concordant in 171 of 180 samples (95%). Both were positive in 123 and both negative in 48 samples. The nine amniotic fluid samples with discrepant results were as follows: Five negative Amniostat-FLM results were associated with trace phosphatidylglycerol by thin-layer chromatography (1% to 2% of total phospholipids), one negative Amniostat-FLM result was associated with 3% phosphatidylglycerol by thin-layer chromatography, and three positive Amniostat-FLM results were associated with negative phosphatidylglycerol by thin-layer chromatography results. There were 14 samples with trace phosphatidylglycerol by thin-layer chromatography; of these, there were nine positive and five negative Amniostat-FLM results. If greater than 2% phosphatidylglycerol by thin-layer chromatography was considered positive, concordance between the two tests was 93% (167 of 180). From these samples, 119 infants were delivered within 72 hours of amniocentesis. There were 100 positive Amniostat-FLM results and all were associated with the absence of respiratory distress syndrome; of the 19 negative Amniostat-FLM results, five were associated with infants who developed respiratory distress syndrome. The present study confirms that Amniostat-FLM is a simple, rapid, and accurate test for determining the presence of phosphatidylglycerol in amniotic fluid.

Agglutination Tests↗

Studies on the interaction of Sendai virus with liposomal membranes. Sendai virus-induced agglutination of liposomes containing glycophorin.

Liposomes constituted with the major sialoglycoprotein of human erythrocytes, glycophorin, were used as models for studies on cell-virus interactions. Liposomes composed of egg yolk phosphatidylcholine, cholesterol and glycophorin were found to interact with the paramyxovirus HVJ to form aggregates. The aggregation process was temperature dependent: it was maximal at 0 degrees C and decreased with increase of the incubation temperature. The activity of viral neuraminidase is also temperature dependent, and it increases with increase of the incubation temperature; release of N-acetylneuraminic acid was negligible at 0 degrees C. Shift-up of the incubation temperature immediately cancelled HVJ-induced agglutination of liposomes. Viruses attached to liposomes seemed to be released into the supernatant when the 'virus-liposome' complex formed at 0 degrees C was incubated at 37 degrees C, possibly as a result of breakdown of the 'binding' site by neuraminidase. The characteristics of the interaction of HVJ with liposomes containing glycophorin appeared to be phenomenologically similar to those of HVJ-cell interaction.

Agglutination↗

A sperm-agglutinating lectin from seeds of Jack fruit (Artocarpus heterophyllus).

A lectin specific for N-acetylgalactosamine was isolated from seed extract of Jack fruit (Artocarpus heterophyllus) by ammonium sulfate precipitation, followed by affinity chromatography on a Affigel-galactosamine-agarose column. The lectin possessed agglutinating activities for human and rat sperm as well as human red blood cells. It was found to have Mr = 62,000 consisting of two dissimilar subunits of Mr = 18,000 and 13,000. It also cross-reacted with an antibody against the lectin of Osage Orange (Maclura pomifera).

Acetylgalactosamine↗

Soluble transferrin receptor-transferrin complex in serum: measurement by latex agglutination nephelometric immunoassay.

The transmembrane protein, transferrin receptor (TfR), exists in serum as a soluble form that lacks cytoplasmic and transmembrane domains (residues 1-100). The level of soluble TfR in serum is a sensitive indicator of total erythropoiesis and iron deficiency. This study revealed that the major part of soluble TfR was saturated by transferrin (Tf) in serum, forming a stable complex which was more immunoreactive than intact TfR. Thus, we proposed that serum soluble TfR should be measured as the TfR-Tf complex (TRC), using prepared TRC for assay standardization. We developed a new assay for TRC, employing antibody-coated latex agglutination nephelometry (LA). Rapid and reproducible measurements were achieved using an automated analyzer. The values obtained by this LA assay were closely correlated with those obtained by conventional enzyme immunoassay (r = 0.967). The mean level of TRC in 179 adult healthy subjects was 1.62 mg/l. Patients with iron-deficient anemia showed significantly higher TRC levels than the healthy subjects.

Adult↗