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PubMed2002

Hemagglutinating factor (HAF) associated with adhesiveness in enteroinvasive Escherichia coli (EIEC).

A cell-associated mannose-resistant hemagglutinating factor (HAF) was extracted from enteroinvasive Escherichia coli (EIEC) serotype O124:H- by sonication. Ultrastructural analysis of EIEC and immunocytochemical assays with the cell-free HAF and EIEC bacterial cells on HeLa cells, suggested that the HAF is a non-fimbrial putative adhesive factor that mediates in vivo adherence of EIEC to human epithelial cells.

Antibodies, Bacterial↗
PubMed1991

The hemagglutinating adhesin HA-Ag2 of Bacteroides gingivalis is distinct from fimbrilin.

We carried out a series of immunoblots with antigenic preparations from the periodontal pathogen Bacteroides gingivalis using antisera of restricted specificity for the hemagglutinating adhesin HA-Ag2, and for the major structural subunit of the fimbriae (fimbrilin). We have been able to show that these 2 antigens are distinct. The fimbrilin subunit had an apparent molecular weight of 42 kDa in all of the bacterial preparations tested. HA-Ag2 occurred as a pair of bands at 43 and 49 kDa in outer membranes prepared as extracellular vesicles, and at 33 and 38 kDa in glass-bead-EDTA extracted antigens and in sheared-cell outer membranes prepared in the presence of EDTA. No HA-Ag2 was found in an enriched fimbrial preparation. The 2 antigens could thus be distinguished on the basis of their behaviour when subjected to different extraction techniques. The lower apparent molecular weight of HA-Ag2 (a pair of bands at 33 and 38 kDa) was invariably associated with the presence of EDTA in the buffers used to prepare the extracts, and the effect could be partially prevented by adding MgCl2 to the extraction buffer. The difference in apparent molecular weight of HA-Ag2 in the different extracts can thus be attributed either to an EDTA-sensitive tertiary conformation of its component polypeptides, or to an EDTA-sensitive linkage of each of these polypeptides to an unknown component of approximately 10 kDa.

Adhesins, Bacterial↗
PubMed1991

Characteristics of hemolytic and hemagglutinating activities of Treponema denticola.

Treponema denticola, a suspected periodontal pathogen, was shown to agglutinate and lyse human red blood cells. Both activities were cell-associated, heat-labile, and produced during the exponential growth phase. Hemolytic activity was time-dependent and required incubation at 37 degrees C; further incubation at 4 degrees C increased the hemolysis. Hemagglutination was reduced in the presence of D-glucosamine, EDTA or sodium salicylate, whereas hemolytic activity was affected by calcium chloride, magnesium chloride, EDTA or sodium salicylate. These biological activities may favor a higher concentration of hemin-containing compounds in periodontal sites, and therefore represent additional potential virulence mechanisms.

Hemagglutination↗
PubMed1975

A new reversed passive hemagglutination test for detection of HBsAg.

A new reversed passive hemagglutination test for HBsAg, termed Raphadex B, has been developed using immunochemically purified chimpanzee anti-HBs bound to stabilized human erythrocytes. The test has been found to have equivalent sensitivity to the Ausria 125I radioimmunoassay, and detected a similar number of HBsAg-containing specimens in screening of volunteer blood donors. This method offers an economical approach to third generation methodology for hepatitis B screening of blood donors.

ABO Blood-Group System↗
PubMed1978

Automatized hemagglutination kinetics.

An automatized technique for the study of hemagglutination kinetics is described. It was used to differentiate between homozygous and heterozygous individuals in the Rh system. Other applications are proposed.

ABO Blood-Group System↗
PubMed1978

Influence of various physicochemical factors on hemagglutination.

A study is made of the influence on hemagglutination of various physicochemical parameters, such as: cell shape, cell distance, extracellular colloid-osmotic pressure, degree of hydration of the cell surface, and cell zeta-potential. To a varying extent these are changed by: the addition of divers soluble polymers, the action of enzymes, the presence of various salt ions, centrifugation, agglomeration at low ionic strength, complex coacervation with cationic polymers, and the interaction with different kinds of antibodies. Among the more important parameters are cell distance and cell shape (particularly when the latter tends to spiculation), while the influence of zeta-potential appears to be of fairly minor consequence. The action of dissolved polymers is mediated through polymer bridging, increase in extracellular colloid-osmotic pressure, decrease of cell surface hydration, and change in cell shape. A peculiarity of dextran is that in addition to all of the above, it causes pronounced erythrocytic spiculation.

Antigen-Antibody Complex↗
PubMed1979

Detection of hepatitis B surface antigen with the miniature centrifugal fast analyzer. A modified reversed passive hemagglutination procedure.

A modified reversed passive hemagglutination test for the detection of hepatitis B surface antigen HBsAg is described. Sera and reagent cells coated with antibody to HBsAg (anti-HBs) are loaded separately into the rotor of a miniature centrifugal fast analyzer. The rotor is centrifuged briefly to transfer the components into its cuvettes. After mixing, the suspensions are allowed to stand at room temperature for 30 min, following which the rotor is again centrifuged and the absorbance of each cuvette is monitored. Cells suspended in serum containing HBsAg leave the light path more rapidly than cells suspended in sera free of antigen. The magnitude of change in absorbance varies directly with the concentration of the antigen. In 45 sera tested by the conventional V-plate technique, findings were as follows: 21 positive, 19 false positive and 5 negative. The automated procedure unequivocally differentiated the 21 positives; results for the false positive and negative specimens were identical and clearly distinguishable from the positive results. The automated procedure enhances specificity, offers equivalent sensitivity, and results that are quantitative and objective.

Absorption↗
PubMed1995

Anti-HLA antibody screening with extracted platelet HLA antigens by the mixed passive hemagglutination method.

For the detection of anti-HLA antibody in the serum of patients receiving frequent platelet transfusions, we developed a new method in which platelet antigens are extracted into physiological saline containing 3% sucrose and coated as a layer on a U-type Terasaki plate. In the present study, screening of anti-HLA antibodies was conducted with this plate by the mixed passive hemagglutination test although the plate contained both HLA and HPA. The reactivity determined by this method correlated well with antihuman immunoglobulin-lymphocyte cytotoxicity test (AHG-LCT) results (r = 0.963). The plate can be preserved for at least 2 years at -80 degrees C and is easier to handle than the frozen lymphocyte panels used in the LCT test in which lymphocytes must be kept alive. This new method is an alternative way to screen HLA antibodies. In addition, it is expected that this method will be used to screen anti-HPA antibodies.

Blood Platelets↗
PubMed2005

Determination of gelatinase A using a modified indirect hemagglutination assay in human prostate cancer screening and assessment of its correlation with prostate-specific antigen parameters.

BACKGROUND: Prostate cancer is the most common malignancy affecting men and is a major cause of cancer death. There are increasing data on novel tumor markers, such as gelatinase A, which play a key role in tissue invasion and metastasis. OBJECTIVES: We designed a study to evaluate total gelatinase A content using a simple and applicable Indirect hemagglutination (IHA) test in harmony with gelatinase A activity in serum samples as compared with prostate-specifc antigen (PSA) parameters. METHODS: In this study, we analysed the circulating form of gelatinase A (MMP-2) in patients suffering from either benign prostate hyperplasia (n=54) or prostate cancer (n=26) versus normal individuals as control (n=26). The gelatinolytic activity was determined by zymography and total MMP-2 content was measured by a novel IHA method. Total PSA and free PSA were quantified using a standard ELISA technique. RESULTS: Correlation of densitometric analysis of gelatinase A activity and IHA titer is significant at the 0.01 level (P<0.01, rho=0.916). Correlation of PSA and IHA titer is significant at the 0.01 level (P<0.01, rho=0.746). Correlation of free PSA and IHA titer is significant at the 0.01 level (P<0.01, rho=0.749). Borderline of IHA titer in patients with prostate cancer was 512+/-1 tube titer, in benign prostate hyperplasia patients was 128+/-1 tube titer and the titer in normal individuals was 8+/-1 tube titer. CONCLUSIONS: These results demonstrate that assessment of gelatinase A might be a promising procedure for monitoring and screening patients with prostate cancer.

Adenocarcinoma↗
PubMed1994

Expression of antigenically distinct fimbriae with hemagglutination and HeLa cell adherence properties by an enteroaggregative Escherichia coli strain belonging to the enteropathogenic serogroup.

An enteroaggregative Escherichia coli (EAggEC) strain (DS92), isolated from a case of infantile diarrhea, was shown to express mannose-resistant hemagglutination and HeLa cell adhering properties when grown at 37 degrees C but not at 28 degrees C. Cellular adherence properties of DS92, which belonged to enteropathogenic serogroup 0125, were shown to correlate well with the expression of fimbriae that were encoded by a 112 kb plasmid. The fimbriae of the EAggEC strain DS92 were composed of 20 kDa subunit proteins and were serologically distinct from fimbrial or non-fimbrial cell surface antigen(s) of other diarrheagenic E. coli strains including the reference EAggEC strain 17-2. Interestingly, the 20-kDa fimbrial protein was found to be antigenically related to 18- and 14.5-kDa cell surface proteins of two other locally isolated EAggEC strains belonging to the enteropathogenic serogroup 086.

Animals↗
PubMed2001

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting.

Tuberculous meningitis (TBM) is one of the commonest chronic infections of the central nervous system (CNS). Diagnosis of TBM has been a problem as it causes various clinical manifestations which can be confused with those of other chronic infections of the CNS such as neurocysticercosis (NCC), neurobrucellosis and cryptococcal meningitis, that are prevalent in many underdeveloped and developing countries. Differential diagnosis of TBM can be made by detecting circulating mycobacterial antigens in CSF by immunoassays. In this study, a reverse passive hemagglutination (RPHA) has been developed using rabbit antimycobacterial IgG for detection of circulating mycobacterial antigens in CSFs from chronic infections of the CNS in order to develop a rapid, simple, sensitive and cost-effective method. Circulating mycobacterial antigens were characterized by immunoblot assay. The sensitivity limit of RPHA was 400 ng ml(-1). RPHA was specific as antimycobacterial IgG did not show any reaction with porcine Cysticercus cellulosae which was used as a control antigen. RPHA could detect mycobacterial antigens in CSF at a sensitivity level of 94.11% with a specificity of 99.0%. Immunoblot analysis of RPHA positive CSFs revealed predominantly 30-32 kDa and 71 kDa antigens whilst 6, 86, 120, 96 and 110 kDa showed varied degree of reactivity. Antigens of masses 30-32 and 71 kDa were absent in culture filtrate of Mycobacterium tuberculosis H37Rv grown in Proskeur-Beck liquid medium. RPHA is a rapid, simple and sensitive immunological method with a long shelf life of 6-8 weeks if stabilized coated erythrocytes are stored at +4 degrees C. RPHA could be used as an additional immunodiagnostic tool in both differential diagnosis and prognosis of TBM. Immunoblot results indicate that 30-32 kDa and 71 kDa antigens are cell wall derived.

Antigens, Bacterial↗
PubMed2002

Assessment of antibody responses to antigens of Mycobacterium tuberculosis and Cysticercus cellulosae in cerebrospinal fluid of chronic meningitis patients for definitive diagnosis as TBM/NCC by passive hemagglutination and immunoblot assays.

Tanned sheep erythrocytes stabilized with pyruvic aldehyde and glutaraldehyde, called double-aldehyde-stabilized cells, were used to standardize passive hemagglutination assay (PHA) for detection of antibody responses to sonicate extract of Mycobacterium tuberculosis and Cysticercus cellulosae soluble antigens. PHA was performed in the following groups of cerebrospinal fluid (CSF) samples: group I - chronic infections of the central nervous system with the possible diagnosis of tuberculous meningitis (TBM), tuberculoma and neurocysticercosis (NCC) (n=88), and group II - controls which included (a) non-infectious non-neurological conditions (n=30), (b) infectious neurological conditions (n=21) and (c) non-infectious neurological conditions (n=133). PHA could detect anti-mycobacterial antibodies at the sensitivity level of 80.76% with a specificity of 92.4% and anti-cysticercal antibodies with a sensitivity of 100% and specificity of 92.94%. However, in 6.33% (i.e. 14/221) of group I and group II (c) CSFs both anti-mycobacterial and anti-cysticercal antibodies were detected. Immunoblot analysis of CSFs derived from TBM patients reacted predominantly to 120-kDa, 96-kDa, 65-kDa, 38-kDa, 26-kDa, 23-kDa, 19-kDa and 12-14-kDa and 4-6-kDa antigens of M. tuberculosis sonicate extract (MTSE), whilst CSFs of proven NCC reacted to >110-kDa, 96-kDa, 80-kDa, 66-68-kDa, 52-kDa and 26-28-kDa antigens of porcine whole cyst sonicate extract (PCSE). On immunoblot analysis, some of the CSFs of TBM patients were PHA positive for both MTSE and PCSE showed antibody reactivity to 70-kDa and 10-kDa antigens of C. cellulosae. Similarly CSF antibody of some Guillain Barre syndrome and myeloradiculopathy patients reacted with cysticercal antigens. But per se no cross-reactivity between MTSE and anti-cysticercal antibodies and vice-versa were observed. However, findings of this study should alert laboratory personnel especially in endemic areas to be extra careful in interpretation of antibody detection results.

Animals↗
PubMed1975

Early detection of hepatoma: prospective study in liver cirrhosis using passive hemagglutination and the radioimmunoassay.

(1) Passive hemagglutination and radioimmunoassay are suitable methods for the detection of AFP in the low concentration range. (2) In 3.72% of the cases a clinically unknown carcinoma was found in an unselected group of patients with liver cirrhosis. (3) 21.9% of the patients showed AFP elevations up to 2000 ng/ml. In 10.6% of this group, increasing titers demonstrated a primary liver cell carcinoma. In 89.4% a transitory rise of AFP was not associated with tumor growth. Levels return to normal values within three months in 90% of the cases. (4) Transitory AFP elevations are not correlated to clinical conditions (praecoma, coma, delirium, bleeding, ascites, shunt) or to biochemical parameters (GOT, GPT, bilirubin, prothrombin complex time, gamma-globulin). (5) A temporary rise in AFP is more frequently observed in groups with high hepatoma incidence than in groups with low hepatoma incidence. (6) Therefore, it may be suggested that a transitory rise of AFP could reflect a "primary reaction" of carcinogenesis. (7) Primary liver cell carcinoma is found to be more frequent in posthepatitic than in postalcoholic, cryptogenic, and other cirrhosis and to be more frequent in australia-antigen positive than in australia-antigen negative cases. (8) Routine serological tumor antigen screening of patients with a precancerous disease is useful.

Carcinoma, Hepatocellular↗
PubMed1993

[Detection of antibody to varicella zoster virus by immune adherence hemagglutination].

Anti varicella-zoster virus (VZV) antibodies were detected by an immune adherence hemagglutination (IAHA) test, and were compared with the CF test, IFA test and ELISA test, respectively. Type O, Rh-positive RBC for IAHA was obtained from five healthy volunteers. All five RBCs had sufficient sensitivity as the indicator cell. Optimum incubation temperature was 37 degrees C in the serum and in the complement. The complement was obtained from guinea pig sera, and the most suitable concentration was 1;100. The convalescent VZV antibody titers were similar to the values obtained from any of the methods previously mentioned. However, mean titers measured by CF were about two-to fourfold lower than in the values of IAHA. Seroconversion rates of the live VZV vaccine, as detected by CF and IFA were relatively low (CF; 76%, IFA; 56%). In contrast, those obtained from ELISA and IAHA showed perfectly (100%). These results indicate that IAHA has sufficient sensitivity and specificity in the detection of VZV antibodies. In addition, the IAHA test is thought to be a rapid and easy test to perform in ordinary laboratories.

Antibodies, Viral↗
PubMed2003

[Measles antibody followed after measles vaccination: comparison with gelatin beads agglutination (PA) antibody, hemagglutination inhibition (HI) antibody and IgG-avidity].

The immunity status of 37 cases has been tested at 6 weeks and 3.5 years after vaccination by using gelatin beads agglutination (PA), hemagglutination inhibition (HI) and avidity of specific IgG. The geometric mean titer (GMT) of HI antibody was 34.5 at 6 weeks and had fallen to 17.9 over 3.5 years. On the other hand, GMT of PA antibody was 36.5 at 6 weeks and had increased to 286.0 over 3.5 years. Also, the average of measles specific IgG avidity was 4.5% at 6 week and had increased to 45.4% over 3.5 years. The multiple regression analysis was performed in order to investigate the relationship of HI antibody and avidity to PA antibody. The significant involvement of the HI antibody and avidity to PA antibody (standardized partial regression coefficients; 0.612, 0.726) was recognized and the multiple correlation coefficient was 0.880 (p < 0.001). Hence it was strongly suggested that the increase of PA antibody with time could be dependent on that of IgG avidity.

Agglutination↗
PubMed1966

Sialic acid binding sites: role in hemagglutination by Mycoplasma gallisepticum.

Hemagglutination of turkey erythrocytes by Mycoplasma gallisepticum was inhibited by mucoproteins containing sialic acid, by sialic acid itself, and by treatment of the erythrocytes with neuraminidase. Neuraminidase treatment of the mucoprotein-rich inhibitors reduced or abolished their inhibitory activity. The findings indicate that sialic acid on the erythrocyte surface provides binding sites for Mycoplasma gallisepticum.

Animals↗
PubMed1971

Factor 8 detection by hemagglutination inhibition: hemophilia A and von Willebrand's disease.

Factor VIII activity was detected immunologically in both the serums and plasmas of 14 normal individuals and 14 patients with hemophilia A. A hemagglutination-inhibition test with rabbit antibody to highly purified (10,000-fold) factor VIII from humans was used. Serums and plasmas from eight patients with von Willebrand's disease showed little or no factor VIII activity in this test, an indication that the test may serve as a specific assay for differentiation between von Willebrand's disease and hemophilia A.

Animals↗