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PubMed1982

Evaluation of slide agglutination and ring precipitation tests for capsular serotyping of Haemophilus pleuropneumoniae.

Rapid slide agglutination (RSA), quantitative plate agglutination, slow tube agglutination (STA), and ring precipitation (RP) tests were performed on 200 isolates of Haemophilus pleuropneumoniae by using the type sera produced in rabbits against five known serotype strains and one strain 202. RSA and RP tests both yielded the same results as those by STA. None of the agglutination procedures could be used for serotyping isolates that autoagglutinated in saline. The RP test was successfully used for serotyping such strains. The specificity of the RSA and RP tests was confirmed by cross-absorption studies. All of the isolates except two had strong serotype-specific activities. The most common serotype isolated in Quebec was serotype 1, followed by serotypes 5 and 2. None of the isolates belonged to serotypes 3 and 4. Only two isolates were found to be untypable; they could possibly belong to serotype(s) not yet defined. The RSA and RP tests may be at least as reliable as the STA test, but easier to perform, less expensive, and much more rapid than any of the other methods reported. Of all the procedures studied by us, the RP test proved to be the method of choice for serotyping H. pleuropneumoniae; hence, it should replace the STA test for serotyping H. pleuropneumoniae.

Agglutination Tests↗
PubMed1983

Aeromonas hydrophila typing scheme based on patterns of agglutination with erythrocytes and yeast cells.

An agglutination typing scheme has been developed for strains of Aeromonas hydrophila. Primary agglutination typing is based on testing agar-grown A. hydrophila cells with human, horse, rat, and guinea pig erythrocytes and Saccharomyces cerevisiae cells. Further subdivision of primary groups is based firstly on whether yeast cell agglutination is inhibited by a D-mannose polymer, yeast mannan, and secondly on patterns of inhibition of hemagglutination by yeast mannan and the monomeric sugars L-fucose, D-galactose, and D-mannose. A total of 320 isolates were tested, and these were divisible into 39 distinct types on the basis of this scheme. Application of this typing scheme in the future to isolates of A. hydrophila known to be associated with human infection may enable correlations to be made between particular agglutination types and human pathogenicity.

Aeromonas↗
PubMed1984

Comparison of indirect hemagglutination and rapid plate agglutination tests with counterimmunoelectrophoresis for typing Pasteurella haemolytica.

A rapid, simple, and accurate counterimmunoelectrophoresis (CIE) technique was developed and compared with the indirect hemagglutination and rapid plate agglutination techniques for serotyping cultures of Pasteurella haemolytica. The CIE test had 100% correlation with the conventional indirect hemagglutination test and, after serum absorption, correctly identified cultures representing the 12 established serotypes and 49 field isolates of P. haemolytica with reasonable rapidity. Cross-reactions were observed in the CIE and rapid plate agglutination tests but not in the indirect hemagglutination test with antisera prepared from the 12 established serotypes. These cross-reactions were eliminated from the CIE test but not from the rapid plate agglutination test by absorption of antisera with cells which possessed the cross-reacting antigens. Avian isolates of P. haemolytica did not type with antisera to the 12 established serotypes by any of the methods. Both homologous and heterologous reactions were observed with these strains in the rapid plate agglutination and CIE tests with antisera prepared from six selected cultures. These results support the previous finding that the taxonomic relationship of these avian strains to P. haemolytica is questionable.

Agglutination Tests↗
PubMed1994

Rapid detection of methicillin-resistant Staphylococcus aureus strains not identified by slide agglutination tests.

Seventy-nine methicillin-resistant Staphylococcus aureus (MRSA) strains, isolated during 1980 to 1990, were classified as MRSA Aggl- (14 strains) and MRSA Aggl+ (65 strains) strains on the basis of test results in slide agglutination assays designed to detect fibrinogen-binding protein (clumping factor) and protein A on the staphylococcal surface. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that lysostaphin digests of MRSA Aggl- strains contained a high-molecular-weight protein which was not detected in digests of MRSA Aggl+ strains. Immunization of rabbits with an MRSA Aggl- strain produced an antiserum which agglutinated all MRSA Aggl- strains and also 64 of 65 MRSA Aggl+ strains. Only 1 of 68 coagulase-negative staphylococci showed agglutination in this assay. The anti-MRSA Aggl- antiserum reacted mainly with a 230-kDa staphylococcal surface protein but also with a 175-kDa protein, probably formed by proteolysis of the former and a few slightly smaller proteins. These could not be immunologically detected in lysostaphin digests of MRSA Aggl+ strains. Purified antibodies reacting with the 230-kDa protein agglutinated all MRSA Aggl- strains, indicating that the protein is located on the surfaces of staphylococci. The results suggest a tentative role for the 230-kDa protein or its fragments as a novel target to develop more efficient rapid identification methods for S. aureus, including MRSA.

Agglutination Tests↗
PubMed1979

Nuclease enhancement of specific cell agglutination in a serodiagnostic test for Neisseria gonorrhoeae.

Antiserum to a purified type R lipopolysaccharide antigen isolated from Neisseria gonorrhoeae was used in a slide agglutination test and compared with conventional carbohydrate utilization and fluorescent antibody tests to confirm the identity of laboratory cultures classified as typical or "atypical" N. gonorrhoeae. Cultures of Corynebacterium vaginalis, N. meningitidis, N. catarrhalis, N. sicca, and N. lactamicus were also tested in the slide agglutination procedure. The addition of both deoxyribonuclease and ribonuclease (1 mg/ml) to the cell suspension medium of phosphate-buffered saline improved the sensitivity and specificity of the agglutination reaction for N. gonorrhoeae. Problems relating to the agglutination test as an aid in identification of N. gonorrhoeae are discussed.

Agglutination Tests↗
PubMed1979

Serological grouping of streptococci by slide agglutination.

Streptococcal grouping sera for groups A, B, C, and G prepared for conventional testing by precipitation were made specific by absorption and used to identify streptococci by slide agglutination with and without staphyloccocal coagglutination. Trypsinised suspensions of 1055 strains, identified by precipitation as belonging to group A, B, C, or G, were tested by slide agglutination. Of these, 998 were correctly identified using a streptococcal suspension and antisera alone and a further 65 were identified when a loopful of protein A-positive staphylococci was added. Suspensions of 88 strains not of groups A, B, C, or G gave no reaction in the agglutination test with or without the addition of staphylococci. Group polysaccharide extracted by conventional methods also caused agglutination of staphylococci on a slide when specific antiserum was added. Growth from primary or secondary cultures digested in streptomyces enzyme for only 15-30 minutes provided an excellent antigen for a quick and simple method of streptococcal grouping using non-sensitised staphylococcal suspension and specific antisera for coagglutination.

Agglutination Tests↗
PubMed1981

Detection of HLA-DRw specificities by mixed agglutination.

By means of mixed agglutination test with staphylococcal protein A indicator system, DRw antigens were studied on lymphocytes obtained from tonsils, peripheral blood and thymuses. Monospecific antisera to DRw 1, 2, 3, 6 and 7 which were preadsorbed with pooled platelets gave mixed agglutination titers of 300 or more against B cells of human tonsilar lymphocytes and peripheral blood lymphocytes. The DR phenotypes of human tonsilar lymphocytes and peripheral blood lymphocytes determined by mixed agglutination tests corresponded to those established by lymphocytotoxicity tests. Upon stimulation of thymocytes or T cells of peripheral blood lymphocytes by Con-A, DRw antigens became detectable by mixed agglutination tests. Simultaneous experiments on the stimulated T cells of peripheral blood lymphocytes by membrane fluorescence tests showed that 21-45% of the stimulated T cells possessed DRw antigens at a detectable level, while less than 2% of unstimulated cells were positive. These results indicated that DRw antigens are present in the subpopulation of T cells which may expand in the response to Con-A stimulation.

Agglutination Tests↗
PubMed1997

Tissue suspension agglutination: a simple method to screen species-specific and organ-specific reactions.

Agglutination tests with preparations of parenchymatous organs were developed. The tissue suspensions were dried at room temperature after they had been spread as a very thin layer on a glass plate, or otherwise, they were lyophilized. The dried preparations were pulverized and then prepared as stable suspensions in saline. The agglutination test was conducted on a slide by mixing one drop of the tested serum at a convenient dilution with one drop of tissue powder suspension. Agglutination in the form of readily discernible clumps could be assessed after 1-10 min. By means of this procedure, species-specific reactions were studied using suspensions of kidneys of various species. Organ-specific reactions were noted with suspensions of brain and thyroid. Agglutination of thyroid powder was observed with rabbit anti-rabbit thyroid sera as well as with many, albeit not all, sera of patients with Hashimoto's disease.

Agglutination Tests↗
PubMed1985

In vitro attachment, salivary agglutination, and surface fibril density of fresh Actinomyces isolates from two distinct oral surfaces.

Whereas attachment specificity among bacterial species is well recognized, the question of specificity among strains of the same species isolated from distinct oral surfaces has not been studied. Twenty strains of Actinomyces, freshly isolated as they re-populated the cleaned teeth and tongues of a male twin pair, were compared for: (1) their ability to attach to saliva-coated hydroxyapatite (SHA), (2) the density of their surface fibrils, (3) their agglutination in saliva, and (4) their ability to agglutinate human erythrocytes. Differences among the strains were mostly along species rather than tissue source lines. The A. naeslundii strains were the least efficient in SHA attachment and had the least number of fibrils, but they were the most strongly agglutinated strains in saliva. The A. viscosus strains, while showing some tissue source differences in hemagglutination, were consistently alike in SHA attachment and fibril density. Salivary agglutination did not correspond with the ability to attach to SHA. Considering the surface characteristics of the Actinomyces species studied, strains of the same species isolated from distinct oral surfaces are apparently more alike than strains of the two closely-related species isolated from the same surface.

Actinomyces↗
PubMed1976

Proliferation and agglutinability of primary and transformed human epithelial cells in culture.

Primary epithelial populations (HAM) were obtained by dissociation of the amniotic membrane stripped from human placentae. Agglutinability of cells from such normal populations and of cells from the transformed epithelial line WISH was then compared using concavanalin A as mediator. Extensive similar studies have previously been reported with cell strains isolated from other species. Freshly dissociated HAM cells from primary cultures agglutinated much less readily than did cells from WISH populations. Furthermore, the former exhibited a drastic decline in agglutinability as a function of time in suspension culture after trypsinization. Short-term exposure (60 h) of HAM cells in monolayer culture to 5-bromodeoxyuridine (BrdU) elicited heightened agglutinability detectable through 22 days in vitro. Addition of the protease inhibitors n-tosyl-L-lysyl-chloromethyl ketone (TLCK) or p-tosyl-L-arginine-methyl ester (TAME) to the culture medium inhibited proliferation of the WISH line by 40--50% while effecting only a 10-15% inhibition of HAM cells. These results also confirm data with other cell species indicating that high proteolytic activity at the surface of transformed cells may be related to the rapid proliferation rate.

Agglutination↗
PubMed1976

Surface morphology and agglutination of lectin-treated human lymphocytes and lymphoblasts.

Two human lymphoblasts (Raji and EB3) and normal human peripheral lymphocytes were exposed to different concentrations of Concanavalin A and wheat germ agglutinin. The lectin-induced agglutination was determined and correlated with lectin-induced changes in the surface morphology of these cells as studied in a scanning electron microscope. Whenever the lectin induced high agglutinability in a cell type, it also invariably had a smoothing effect on the cell surface. In contrast, when cells did not agglutinate well with a certain lectin, their cell surface remained essentially rough (villous) after addition of the lectin. The correlation found between increased agglutinability and altered cell surface morphology upon treatment with certain lectins suggests that both phenomena result from one and the same process. Additional evidence for this postulate is presented.

Agglutination↗
PubMed2001

Immunological analysis of agglutination in Dirofilaria immitis microfllariae.

The mechanism of agglutination phenomenon of Dirofilaria immitis microfilariae was analyzed. Circulating microfilariae were collected from a D. immitis-infected microfilaremic dog and cultured in the several kinds of sera from dogs and animals. The agglutination of D. immitis microfilariae is a specific phenomenon due to some immune complexes formed with the anti-microfilarial antibody, heat-instable factor(s) and excretory-secretory products of microfilariae. Only live microfilariae were agglutinated and the agglutinated microfilariae remained alive as long as 27 days in culture in vitro.

Agglutination↗
PubMed1976

Serum-induced head-to-head agglutination of bovine spermatozoa.

Addition of cow serum to diluted bull spermatozoa induced marked head-to-head agglutination which declined during incubation for 9 hr at 37 degrees C. Maintenance of the acrosomal cap during incubation was not adversely affected by serum. Agglutinating cells were those with intact acrosomal and cell membranes, as determined by differential interference-contrast and electron microscopy. Single cells in the same treatment groups experienced more rapid acrosomal deterioration than did untreated spermatozoa. Ultrastructurally, agglutinated cells were associated by a close apposition of the cell membranes in the acrosomal region. Structural integrity of the cell membrane and acrosome of agglutinated cells was the same after 9 hr of incubation as at the onset of the incubation period.

Acrosome↗
PubMed1982

Effects of amine substances on susceptibility of cells to agglutination by concanavalin A and on paracrystal formation by vinblastine in untransformed 3T3 cells.

The effects of a secondary amine (ketamine), tertiary amines (dibucaine, lidocaine, marcaine, propanidid, diazepam and chlorpromazine) and a quaternary amine (tetraethylammonium bromide, TEA) on mouse 3T3 cell agglutination by concanavalin A (Con A), on patch formation of Con A receptors on the cell surface, and on paracrystal formation by vinblastine in cytoplasm were studied. These amines enhanced the cell agglutination at low concentrations of Con A, as did the mixture of colchicine and cytochalasin B. Ca++, applied extracellularly, inhibited the effects of these amines on cell agglutination by Con A. The patch formation of Con A receptors on the cell surface as revealed by fluoresceinated Con A was enhanced by these amines. Ketamine, dibucaine and TEA inhibited the paracrystal formation in cytoplasm, as did Ca++ ionophores such as A-23187 and X-537-A. These results suggest that the amines tested affect the fluidity of Con A receptors by impairment of cell membrane structural proteins and result in the increase of the susceptibility of cells to agglutination by Con A.

Agglutination↗
PubMedDate not supplied

Detection of Cryptosporidium parvum antigen by co-agglutination test and ELISA.

Confirmation of the presence of Cryptosporidium in environmental samples is laborious, costly and often difficult. We report here a simple and economic slide agglutination test (co-agglutination test) for detecting cryptosporidial antigen in stool, serum and water. The results show that as a screening method co-agglutination is clearly superior to enzyme-linked immunosorbent assay (ELISA) and modified Ziehl-Neelsen staining, although ELISA is more accurate. The co-agglutination test is recommended for application as a new tool for detecting cryptosporidial antigen in large-scale epidemiological surveys.

Acute Disease↗
PubMed2002

[BjCHI1 from Brassica juncea displays both chitinase and agglutination activity].

The proteins encoded by the Brassica juncea chitinase gene BjCHI1 and its derived genes BjCHI2 and BjCHI3 were expressed by Multi-copy Pichia expression system. The chitinase activity of FPLC purified BjCHI1, BjCHI2 and BjCHI3 were tested and the results showed that all the three proteins degraded both CM-chitin-RBV and colloidal chitin. The Km values of BjCHI1, BjCHI2 and BjCHI3 for CM-chitin-RBV were estimated as 0.799 mg/mL, 0.544 mg/mL and 0.793 mg/mL, respectively. When the colloidal chitin was used as substrate, the Km values were 0.281 mg/mL, 0.388 mg/mL and 1.643 mg/mL, respectively, indicating chitin-binding domain can increase affinity of chitinase to insoluble substrate. In the agglutination activity assay, only BjCHI1 shows activity when the protein concentration was more than 33 micrograms/mL, while BjCHI2 and BjCHI3 without agglutination activity even when the concentration was increased as high as 800 micrograms/mL. This means that the two chitin-binding domains in BjCHI1 are essential for agglutination and BjCHI1 is the first protein which shows both chitinase and agglutination activity identified so far in plants.

Agglutination↗
PubMedDate not supplied

[The agglutinability of cholera O-monoclonal immunoglobulins].

A set of 10 monoclonal antibodies specific for vibrio species and of 5 monoclonal antibodies specific for serovar (Ogawa) was studied. These monoclonal antibodies were directed toward V. cholerae O1 antigen in agglutination reaction and on slide plates. Monoclonal antibodies agglutinating typical strains of cholera vibrios with titration range from 1: 6000 to 1: 25,000 were selected. MA were revealed to interact with cholera vibrio strains with reduced agglutinability. The set of agglutinating O monoclonal immunoglobulins was developed for laboratory identification of cholera O1 vibrios.

Agglutination Tests↗
PubMed1992

The differentiation between freshwater and marine fish trypanosomes by lectin agglutinability.

The agglutinability of three fish trypanosomes, Trypanosoma boissoni and T. triglae senegalensis from marine fish, and T. carassii from freshwater fish was compared. The tests were performed with trypomastigotes and epimastigotes in the exponential and stationary growth phases. The simple agglutination test, performed in microwell plates, used ten purified lectins, and revealed a clear difference between marine and freshwater strains when compared for their agglutinability with the HPA lectin. Whereas the cells of T. boissoni and T. triglae senegalensis became strongly agglutinated by this lectin, those of T. carassii remained unagglutinated in the same conditions. However, the cells of T. triglae senegalensis reacted positively with ConA lectin in the stationary phase only. The differences in structure and composition of surface polysaccharides stable after prolonged cultivation in vitro are inferred on the basis of the results obtained.

Agglutination Tests↗