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PubMed1978

[Agglutination and acrosome reaction of ram spermatozoa incubated in the bovine folicular fluid].

Fresh-ejaculated sperm of ram was incubated at a temperature of 38 degrees C in the bovine follicular fluid and homologous blood serum (blood plasma). The spermatozoa were studied in native state under a microscope with phase contrast and as ultra-thin sections under a transmission electron microscope. In the follicular fluid and in the blood serum, strong agglutination of spermatozoa occurred, with its maximum after about two hours. Only the heads of spermatozoa agglutinated, the flagella being loose were arranged parallelly. Although the progressive motility of spermatozoa was not observed, the motility of flagella was not affected. The investigation under the electron microscope showed that the agglutination occurred only in acrosomes of intact spermatozoa. It was also found out that the follicular fluid induced the acrosome reaction of spermatozoa. The course of the acrosome reaction is similar to that in the other mammals: first of all plasma membrane becomes undulated, then it fuses with the outer acrosome membrane, giving origin to vesicles within the entire acrosome, except the equatorial segment. The acrosome reaction was found in about 5% spermatozoa, and therefore it may be assumed that secretions of the oviduct and uterus play their role to induce the acrosome reaction.

Acrosome↗
PubMed1975

[Microtiter agglutination test for salmonella and brucella antibodies (author's transl)].

To compare the tube agglutination test and the microtiter plate agglutination procedure, 207 human sera were tested for agglutinins to Salmonella and 267 sera for agglutinins to Brucella by both methods. A positive reaction in the microtiter test was indicated by a smooth or irregular mat shape at the bottom of each well in the microtiter plate, whereas a negative reaction was shown by a settling of the antigen to a button shape. As can be seen in table 1, identical titer or only one dilution difference between both tests were found in more than 93% of the samples tested. The plate test was easier to perform and saved time, space, antigen and serum. The microtiter-agglutination technique can, therefore, be recommended for the routine laboratory for serology.

Agglutination Tests↗
PubMed2003

[Accuracy of the slide agglutination method evaluated with PCR in typing Haemophilus influenzae isolates].

In 1998, the Colombian government initiated an immunization program for children under one year of age with a Haemophilus influenzae capsular type b conjugate vaccine. After two years, the surveillance program of the Colombian Instituto Nacional de Salud Microbiology Group reported a 40% decrease in meningitis cases caused by H. influenzae. This effect was attributed to the vaccination. The surveillance program uses the standardized slide agglutination technique to serotype H. influenzae. The current study validated the accuracy of the slide agglutination method by means of the PCR technique. From children under five years of age, 146 isolates were obtained. These were collected between 1999 and 2002, and were characterized by biochemical tests and serotyped by the INS as part of the surveillance program. PCR confirmed 93% of the H. influenzae serotype b and 92% of the other serotypes. When the slide agglutination technique is conducted under a strict quality control program, it remains a sensitive and specific tool for serotyping H. influenzae.

Agglutination Tests↗
PubMed1975

[Normal "irregular" hemagglutinins in the sera of healthy adults found by using increased volumens of serum in agglutination and conglutination tests (author's transl)].

Using increased volumens of serum [in the Multiple-Dosage (MD) or a High Dosage (HD) test] with a sensitive manual reading technique all sera of 37 donors of blood group A2 showed irregular anti-A1(alpha1) antibodies. With the same technique, up to 1/3 of the sera of 417 healthy blood donors agglutinated red cells of a selected antigen pattern at room temperature; at 4 degrees C the ratio of positive reacting sera was much higher. The specificity of the antibodies, as indicated by the antigramme type, could be confirmed in the most cases by reacting with a series of known positive and negative reference red cells. The eluates of some of the sera yielded the same antigramme pattern as the native sera did. Some of the antibodies showed a specificity corresponding to a red cell property of the respective blood donor; but not in each case the antibodies were capable to agglutinate the donor's cells. If "autoagglutinins" were strong, frequently they revealed a character of panagglutinability, particular in the cold. With the antiglobulin test, which followed the NaCl-MD procedure, the number of positive reactions increased; on the other hand, there were some of the previously positive agglutination tests, which became negative during the course of the AHG step.

ABO Blood-Group System↗
PubMed1954

Proposed standard agglutinating sera for typhoid and paratyphoid A and B fevers.

The preparation of seven Standard Agglutinating Sera considered suitable to serve as international standard preparations for use in the serodiagnosis of typhoid and paratyphoid infections are described, and proposals are made for their further examination on a collaborative international basis.Details are given of antigenic materials and immunizing procedures which should suffice to enable substandards, based on the proposed International Standards, to be prepared locally.With the aid of the proposed Standard Agglutinating Sera it is possible to determine the "Standard Agglutinin Titre" of a serum under examination by any technique of the agglutination test, provided that a suspension of average sensitivity is employed.

Agglutination Tests↗
PubMed2003

[Experimental study on hybridoma cells agglutination test for diagnosis of schistosomiasis japonica in mice].

OBJECTIVE: To detect the infection of Schistosoma japonicum in mice with a novel test based on agglutination of hybridoma cells and to study the mechanism of the hybridoma cells agglutination. METHODS: The procedure was developed with a murine cell line H226 producing a monoclonal antibody specific to schistosome 31/32 kDa antigen and sera collected from mice infected with different numbers (10, 30, 50) of S. japonicum cercariae in different period. Immunofluorescent test was carried out with the hybridoma cells and schistosome-infected sera. RESULTS: The circulating antigen was detected by the test as early as 2 weeks after a heavy infection and all mice showed positive results in the test by 5 weeks after infection. The titers of antigen rose along with the time post infection, and the titers of sera from heavy infection were statistically higher than that from the mice receiving a lower number of cercariae. Specific yellowish green fluorescence appeared on the membrane of the hybridoma cells; no signal was detected inside. CONCLUSION: Hybridoma cell agglutination test (HCAT) may become useful to diagnose schistosomiasis.

Agglutination Tests↗
PubMed1992

Use of beef extract as a diluent in direct agglutination test for visceral leishmaniasis.

Direct Agglutination Tests (DAT) have been developed for sero-diagnosis of visceral leishmaniasis (VL). The objective of this study was to find a more suitable diluent for serum dilution in a Direct Agglutination Test. 1% of beef extract (BEX) was used as a diluent in the Direct Agglutination Test; 1% foetal bovine serum (FBS) and 0.2% gelatine (Gel) were included for comparison as alternative diluents. Serum from VL patients, individuals suffering from hydatidosis, syphilis and from healthy adult Kenyans was used as control. Beef extract was found to be easy to handle, less expensive and results comparable to those obtained from the other diluents.

Agglutination Tests↗
PubMedDate not supplied

[PCR-based capsular typing of Haemophilus influenzae isolates non-typeable by agglutination].

Haemophilus influenzae is recognized as a pathogenic agent responsible of localized and systemic infections. Six antigenically different capsular polysaccharide types have been described (a, b, c, d, e, and f ) which can be identified by slide agglutination with specific antisera. Besides there are non capsulated strains that cannot be typed by slide agglutination. The introduction of the conjugated vaccine produced an important reduction of invasive diseases caused by H. influenzae type b. Capsular typing by PCR is the most appropriated method for distinguishing non capsulated strains from capsule deficient type b mutants (b-) and for detecting strains of other serotypes that cannot be detected by slide agglutination. Capsular genotype was studied in 38 isolates of non-typeable Haemophilus influenzae received at INE-ANLIS "Dr. Carlos G. Malbrán" between 2002-2004. Of the isolates included in this study 78.9% of them were recovered from blood cultures and most of them were associated with a respiratory focus. By PCR technique 100% of the isolates were identified as non-capsulate H. influenzae and genotype b-was not detected.

Agglutination Tests↗
PubMed2006

[Agglutination as screening test in routine diagnostic of listeriosis].

INTRODUCTION: The persons with weakened immunity, newborn infants and pregnant women are very much sensitive on infection with bacteria Listeria monocytogenes. Listeria can readily invade the placenta and precipitate premature labor and fetal death. To confirm diagnosis is very difficult because listeriosis is disease very similar to the others acute diseases. Serum threshold values above 1: 320 confirm think about listeriosis when the relevant clinical symptoms are present. Of greater diagnostic relevance is the detection of a rise in titer. Therefore, it is always desirable to test a second sample (collected 2 to 3 weeks later) together with the first sample (stored in the deep-freeze). AIM: The aim of the study is to confirm the advance agglutination as a screening test in routine diagnostic Listeriae monocytogenes. MATERIAL AND METHODS: In testing were included 60 women in the reproductive age to divide in experimental group and control group. By agglutination serologic methods we researched if there is any antibody on Listeria monocytogenes with Listeria antigen. RESULTS AND DISCUSSION: From a total of 60 patients positive samples were 18 (60.00%) in experimental group (30) and 8 (26.70%) in control group (30). CONCLUSION: due to simple way of performing screening test of agglutination the same one are giving adequate antibiotic therapy in a case of positive serological response.

Adolescent↗
PubMedDate not supplied

[Agglutinating and immobilizing activity of an ethanol extract of Bursera fagaroides on human and other mammalian spermatozoa].

Human spermatozoa and those obtained from mouse epididymis incubated in ethanolic from mouse epididymis incubated in ethanolic extracts of the cortex of Bursera fagaroides became agglutinated and immobilized in 100% with 30% of viability decrease. This double effect occurred in 50% of pig'ssperm and 50% of rabbit sperm when the concentration was increased tenfold. The pattern of progressive motility was affected 80% of rat's spermatocytes without agglutination. The values remained the same when the extracts were partially purified with salts of Zn++ and Ba++ or by filtration in a sephadex g-10 column. Analysis with thin layer chromatography of both purified extracts produced 3 compounds in comparison with the ethanolic unpurified extract. The extract of the plant Bursera fagaroides contains 3 compounds, which apparently are glycosides with a potent activity upon agglutination -immobilization and a low effect upon spermatocytes viability, which might be used as contraceptives .

Animals↗
PubMedDate not supplied

[Detection of anti-Brucella spp. antibodies in swine by agglutination techniques and indirect ELISA in the Buenos Aires and La Pampa provinces, Argentina].

Porcine brucellosis is one of the most important zoonoses in this country. Currently, there is no control program for porcine brucellosis in Argentina and the epidemiological situation is still unknown. The purpose of our study was to detect anti-Brucella spp. antibodies in swine in the southwest of the Buenos Aires province and the east of the La Pampa province. Blood samples were obtained when animals were slaughtered. The presence of anti-brucella antibodies was studied by the buffered plate agglutination test (BPA), the tube agglutination test (SAT), the 2-mercaptoethanol (2-ME) agglutination test and indirect ELISA tests, using the cytosolic fraction from Brucella abortus S19 (CYT), and lipopolysaccharide (LPS)-free cytosolic proteins (CP). Out of a total of 325 samples analyzed, 17.8% reacted positively to BPA, 13.8% to SAT, 8.0% to 2-ME, 21.0% to ELISA-CYT and 10.0% to ELISA-CP. These results agree with the few data available in our country and suggest that brucellosis screening should be extended to other regions.

Agglutination Tests↗
PubMed1991

Identification of Mycoplasma gallisepticum by use of monoclonal antibody in a rapid slide agglutination test.

Monoclonal antibody (MAb) against Mycoplasma gallisepticum strain PG31 was produced in BALB/c mice. The MAb (designated M9) was of IgG3 isotype and reacted with an epitope in M gallisepticum antigens with molecular weights of 35, 90, 95, and 98 kilodaltons (kDa). The M9 reacted with M gallisepticum antigens in the dot-blot ELISA and in western blot assays. It agglutinated M gallisepticum strains PG31, F, R, S6, A5969, and 9 field isolates from various sources. A coagglutination assay, using Staphylococcus aureus (Cowan strain 1), was developed to enhance the agglutination of some weakly agglutinating M gallisepticum isolates. The M9 did not react with M synoviae, M iowae, M meleagridis, M gallinarum, or M gallinaceum in any of the aforementioned assays. This MAb may be useful in facilitating laboratory diagnosis of M gallisepticum infections.

Agglutination Tests↗
PubMed1991

[Critical study of ELISA technique and high sensitivity direct agglutination test for the screening of antitoxoplasma IgG].

The authors have evaluated an ELISA (Toxo-IgG EIA; bioMérieux) and a direct agglutination test using a sensitized antigen (Toxo-Screen DA; bioMérieux), with a fluorescent test antibody (Toxo-Spot IF; bioMérieux) as a reference for the screening of toxoplasmosis IgG antibodies. A good correlation was obtained for these two methods with fluorescent test antibody (p less than 0.00001) on 841 sera. However, results obtained with direct agglutination test were better than those obtained with ELISA (direct agglutination versus fluorescent test antibody: 97.4% of agreement; ELISA versus fluorescent test antibody: 96.1% of agreement). In addition, the use of the Receiver Operating Characteristic (ROC) curve, showing the changes of sensitivity and specificity as a function of the possible values of each threshold, indicated that sera with titers between 15-20 IU/ml should be interpreted very cautiously.

Adult↗
PubMed1991

[Role of bacterial agglutination in wound infection].

Method of study of bacterial agglutination by means of serum taken from patients with wound infections was suggested. It was found out that agglutination suppresses viability of microbial cells in the center of agglutinate and reduces the risk of bacterial spreading from the wound over the body.

Agglutination↗
PubMed1991

Evaluation of three slide agglutination tests for rapid identification of Staphylococcus aureus.

Three slide agglutination tests for identification of Staphylococcus aureus were compared. The agglutination tests used for evaluation were Staphaurex (Wellcome Diagnostics), Staphyslide-Test (BioMerieux), and ANI S. aureus TEST (Ani Biotech Oy). A total of 347 isolates were analyzed, including 288 strains of S. aureus, 49 of S. epidermis, 11 of S. intermedius, 12 strains of other staphylococci and 14 non-staphylococcal strains. One hundred of the S. aureus strains were isolates from cases of food poisoning, 129 from mastitis and 59 from other clinical cases. The sensitivities of the tests were also compared using diluted suspensions of S. aureus strains and with purified Protein A dilutions. The results showed that the sensitivities of the tests were 98.6%, 97.9% and 99.0% for Staphaurex, Staphyslide-test and ANI S. aureus TEST, respectively. The specificities were 100% for the Staphyslide test and 98.8% for both the ANI S. aureus TEST and the Staphaurex test. The sensitivities measured with diluted S. aureus strain suspensions and Protein A solutions were equal with the Staphaurex and ANI S. aureus TEST. All the agglutination tests studied proved to be practical, easy to use and accurate for the rapid identification of S. aureus strains from culture isolates.

Agglutination Tests↗
PubMed1978

Similarities in the membrane fluidity of 3T3 and SV101-3T3 cells and its relation to concanavalin A- and wheat germ agglutinin-induced agglutination.

Intact, viable ultransformed 3T3 and transformed SV101-3T3 cells were labeled with fatty acid spin labels and with 2,2,6,6-tetramethylpiperidine-1-oxyl in order to measure the fluidity properties of membrane lipids. Both cell types were grown in regular calf serum and in a lipid-depleted serum supplemented with either oleate or elaidate. The temperature dependence of the spectra obtained revealed inflections that correlate with the temperature below which agglutination with concanavalin A is inhibited, and another inflection that correlates with the temperature below which agglutination with wheat germ agglutinin is inhibited, suggesting that (a) the lipid phase(s) in the vicinity of the receptor(s) for these two lectins differ, and (b) a fluid membrane in the vicinity of the lectin receptor(s) is necessary for agglutination with either concanavalin A or wheat germ agglutinin. Studies with a partially characterized plasma membrane fraction suggest that the plasma membrane fluidity parameters closely resemble those of the intact cell. 3T3 and SV101-3T3 cells show virtually identical fluidity profiles by all of the tests we have applied.

Agglutination↗
PubMed1990

Agglutination of rabbit platelets in plasma by the thrombin inhibitor D-phenylalanyl-L-prolyl-L-arginyl chloromethyl ketone.

The specific thrombin inhibitor D-phenylalanyl-L-prolyl-L-arginyl chloromethyl ketone (FPRCH2Cl, PPACK) has been used as an anticoagulant with human blood to permit the preparation of platelet-rich plasma in which the physiological concentration of Ca2+ is maintained. When we attempted to use FPRCH2Cl (40 microM) as an anticoagulant for rabbit blood, clotting was prevented for at least 6 h at room temperature, but nearly all the platelets agglutinated. Thromboxane B2 was not formed and indomethacin and prostaglandin E1 were not inhibitory. The presence of citrate delayed this agglutination, but only EDTA (not EGTA) prevented it, indicating a dependence on Mg2+. Addition of FPRCH2Cl to suspensions of rabbit platelets in Tyrode-albumin solution had no effect on responses to ADP or collagen, but completely blocked all responses to human or rabbit thrombin. Thus plasma is required for the agglutination. Intravenous injection of FPRCH2Cl (5 mg/3 kg rabbit) reduced the platelet count to less than 50% by 15 to 25 min. Thus FPRCH2Cl cannot be used as an anticoagulant for rabbit blood when platelets are to be examined, and has a profound effect on the platelet count in vivo in rabbits. These effects should be considered when FPRCH2Cl is used as an antithrombotic agent for in vivo experiments with rabbits.

Agglutination↗