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PubMed1988

Solid-phase immunosorbent technique for rapid detection of Rift Valley fever virus immunoglobulin M by hemagglutination inhibition.

A solid-phase immunosorbent technique (SPIT) was adapted to detect Rift Valley fever (RVF) virus-specific immunoglobulin M (IgM) in serum samples from humans vaccinated with Formalin-inactivated RVF vaccine. Microdilution plates coated with goat anti-human IgM were successively incubated with serum samples from human vaccinees, RVF virus hemagglutinating antigen, and goose erythrocytes. The RVF virus-specific IgM in the serum samples from vaccinees bound to the RVF virus antigen and inhibited hemagglutination of goose erythrocytes. SPIT was compared to the IgM capture enzyme linked immunosorbent assay (ELISA) and the indirect immunofluorescent-antibody (IFA) assay and was found to be sensitive in detecting RVF virus-specific IgM antibody, with high correlations between SPIT and the other two tests (Pearson's correlation coefficient [r] = 0.9 and 0.6, respectively). Results of SPIT were obtained within 5 h, offering speed over ELISA (8 h). In addition, SPIT does not require sophisticated equipment or expensive reagents. Serum rheumatoid factor did not produce false-positive reactions in SPIT as in the indirect immunofluorescent-antibody assay and IgM capture ELISA.

Antibodies, Viral↗
PubMed1990

Evaluation of a modified complement fixation test and an indirect hemagglutination test for the serodiagnosis of melioidosis in pigs.

A complement fixation test modified by the addition of porcine serum and an indirect hemagglutination test were used to detect antibodies to Pseudomonas pseudomallei in pigs. These tests together with cultural examinations were carried out with 250 pigs. The sensitivity and specificity values were 79.3 and 99.5% and 82.8 and 93.2% for the modified complement fixation and hemagglutination tests, respectively. When results from the combination of both tests were considered, the values were 86.2 and 92.8%, respectively.

Animals↗
PubMed1992

Comparison of different approaches to measuring influenza A virus-specific hemagglutination inhibition antibodies in the presence of serum inhibitors.

The A/Los Angeles/2/87 (H3N2) (A/LA/2/87) virus is sensitive to inhibitors of hemagglutination present in certain human sera. It was found that the effect of these inhibitors could be removed by treating sera with high-concentration receptor-destroying enzyme or trypsin-periodate or by using inhibitor-resistant viruses in the hemagglutination inhibition (HAI) test. Inhibitor-resistant viruses were not effective for detecting rises in antibody titers in the sera of volunteers infected with the A/LA/2/87 wild-type virus, while rises in antibody titer were readily detected in sera treated with trypsin-periodate and tested against A/LA/2/87 wild-type virus in an HAI test. It is therefore suggested that chemical or enzymatic methods be used to inactivate serum inhibitors and that standard virus be used in the HAI test for the currently circulating H3N2 viruses.

Antibodies, Viral↗
PubMed1992

Indirect hemagglutination assay for diagnosis of Escherichia coli O157 infection in patients with hemolytic-uremic syndrome.

An indirect hemagglutination assay consisting of sheep erythrocytes coated with lipopolysaccharide (LPS) from Shiga-like toxin-producing Escherichia coli O157 was used for the serological diagnosis of E. coli O157 infections in children with classical (enteropathic) hemolytic-uremic syndrome (HUS). One week after the onset of diarrhea (acute phase of the disease), the E. coli O157 antibody titer was greater than or equal to 1:4,096 in 22 of 27 patients with HUS, compared with 4 of 249 controls, the majority of whom had O157 antibody titers of between 1:4 and 1:256. This antibody response was observed in HUS patients with stool cultures positive and negative for E. coli O157. Selective absorption with homologous LPS and heterologous LPS showed that the antibody response was specific for E. coli O157. Because of its simplicity and ease of interpretation, the indirect hemagglutination assay described in this paper is recommended for the serological diagnosis of E. coli O157 infections in patients with HUS.

Antibodies, Bacterial↗
PubMed1998

Evaluation of the indirect hemagglutination assay for diagnosis of acute leptospirosis.

Serology plays an important role in the diagnosis of leptospirosis. Few laboratories have the resources and expertise to perform the microscopic agglutination test. There is a need for rapid and simple serological tests which facilitate the early diagnosis of leptospirosis, while antibiotic therapy may be most effective. A commercially available indirect hemagglutination assay (IHA; MRL Diagnostics, Cypress, Calif.) was evaluated with multiple serum specimens from 107 patients being investigated for leptospirosis. By using a combination of enzyme-linked immunosorbent assay (ELISA) methods for immunoglobulin M (IgM) and IgG antibodies and the microscopic agglutination test, 54 patients were found to have leptospirosis and 53 were found not to have leptospirosis. The sensitivity of IHA for the detection of acute leptospirosis was 100%, the specificity was 94%, the positive predictive value was 95%, and the negative predictive value was 100%. IHA was negative when 13 antinuclear antibody-positive sera, 24 serum specimens from patients with syphilis, and 16 serum specimens false positive by the Venereal Disease Research Laboratory test were tested. IHA was shown to detect both IgM and IgG classes of antibodies in human sera. Serum specimens from 27 dogs investigated for leptospirosis were studied: 3 samples gave nonspecific hemagglutination, but for all remaining samples, the results of IHA and an IgM ELISA were concordant. Performance of IHA was simple, and IHA requires no specialized equipment. It represents a useful assay for laboratories which require a leptospiral diagnostic capability but lack the expertise to perform specialist investigations.

Acute Disease↗
PubMed2000

Comparison of antibody titers determined by hemagglutination inhibition and enzyme immunoassay for JC virus and BK virus.

A comparison of antibody titers to JC virus (JCV) or BK virus (BKV) was made by hemagglutination inhibition (HI) and enzyme immunoassay (EIA) with 114 human plasma samples. Antibody titers to JCV or BKV determined by HI were lower than those determined by EIA. Nevertheless, as HI titers increased so did EIA titers. When antibody data were compared by the Spearman rank correlation test, highly significant correlations were found between HI and EIA titers. Results obtained by plotting EIA antibody titers for JCV against those for BKV generally showed a reciprocal relationship, i.e., samples with high antibody titers to JCV had lower antibody titers to BKV and vice versa. Some samples, however, had antibody titers to both viruses. Of the samples tested, 25.4% (25 of 114) had HI and EIA antibody titers to JCV and BKV which were identical or closely related. This is not the scenario one would expect for cross-reactive epitopes shared by the two viruses, but one suggesting that these samples were from individuals who had experienced infections by both viruses. Adsorption with concentrated JCV or BKV antigen of sera with high antibody titers to both JCV and BKV and testing by JCV and BKV EIA gave results which support this conclusion. Although 52.6% (51 of 97) of the samples from the Japanese population tested had very high antibody titers (>/=40,960) to either JCV or BKV, none of the samples were found by a dot blot immunoassay to have antibodies which cross-reacted with simian virus 40. The results from this study, in agreement with those of others, suggest that humans infected by JCV or BKV produce antibodies to species-specific epitopes on their VP1 capsid protein, which is associated with hemagglutination and cellular binding.

Antibodies, Viral↗
PubMed1976

Rubella hemagglutination-inhibition test: false-positive reactions in sera contaminated with bacteria.

We have shown that bacterial contamination of sera can have a marked influence on the results of the rubella hemagglutination-inhibition (HI) test. In addition to increasing the levels of nonspecific agglutinins, a number of common bacterial species tested had a significant effect on the HI titers. When sera free from rubella-specific antibodies were contaminated with Bacillus subtilis or Pseudomonas fluorescens, HI titers ranged from 16 to 128 (expressed as the reciprocal of the highest serum dilution completely inhibiting hemagglutination) after treatment with heparin/MnCl2. Our observations demonstrate, therefore, that bacterial contamination can be one of the causes of false-positive reactions in this test.

Bacillus subtilis↗
PubMed1978

Type-specific indirect hemagglutinating antibody in patients with Pseudomonas aeruginosa Infection.

Lipopolysaccharide antigens were extracted from heated cell extracts of several serotypes of Pseudonomas aeruginosa. Indirect hemagglutination with the extracts indicated specific reactivity with sera from rabbits immunized with homologous serotypes of P. aeruginosa. Sera from healthy adults and patients infected with P. aeruginosa were studied subsequently and shown to possess antibodies against P. aeurginosa. In patients infected with P. aeruginosa type E, indirect hemagglutination antibody against type E was resistant to 2-mercaptoethanol (2-ME) and classified as immunoglobulin G. In patients infected with any other type of P. aeruginosa and in healthy adults, it was sensitive to 2-ME and classified as immunoglobulin M. The antibody in the seven patients infected with P. aeruginosa was titrated during the course of infection with lipopolysaccharide antigen prepared from the infecting strain. As a result, all patients either possessed 2-ME-resistant antibody or showed antibody rise to homotypic antigen during the infection. However, no patient showed 2-ME-resistant antibody against hetero-typic lipopolysaccharide antigens. In hospitalized patients, the incidence of anti body against type E, G, and I Pseudomonas strains was greater than that against type A. In particular, 2-ME-resistant antibody to all four serotypes was detected at a higher rate in hospitalized patients than in healthy adults.

Antibodies, Bacterial↗
PubMed1979

False-positive reactions in the rapid plasma reagin-card, fluorescent treponemal antibody-absorbed, and hemagglutination treponemal syphilis serology tests.

Sera from 628 nonsyphilitic individuals were tested with the Rapid Plasma Reagin-Card, Fluorescent Treponemal Antibody-Absorbed, and Hemagglutination Treponemal Test for Syphilis tests to ascertain the comparative specificity of these tests. Many sera were also tested with the quantitative Venereal Disease Research Laboratory test. Sera included in the study were from both normal individuals and patients with a variety of illnesses and conditions. The Hemagglutination Treponemal Test for Syphilis gave the lowest overall percentage of false-positive reactions (1.6%), followed by the Fluorescent Treponemal Antibody-Absorbed test (3.3%) and the Rapid Plasma Reagin-Card test (10.8%).

Adult↗
PubMed1974

Hemagglutination-inhibition: rapid assay for neuraminic acid-containing viruses.

Influenza virus particles bind rapidly to vesicular stomatitis, Sindbis, or Rauscher murine leukemia virus particles, forming mixed aggregates demonstrable by electron microscopy. The normal hemagglutinating property of influenza virus is inhibited by these viruses, providing a rapid quantitative assay. Prior treatment with neuraminidase blocks the ability of other viruses to inhibit influenza virus hemagglutination.

Animals↗
PubMed1974

Kinetics of inhibition of influenza virus hemagglutination by homologous antihemagglutinin and antineuraminidase antibodies as measured by means of the photometric antibody concentration unit method.

The kinetics of the reactions of antihemagglutinin (AH) and antineuraminidase (AN) antibodies with homologous influenza virus were examined by use of a photometric hemagglutination inhibition test (antibody concentration unit [ACU] test). The following results were obtained. (i) The isotherms describing the reaction of AN antibodies with homologous virus were found to have much steeper slopes than did the corresponding AH antibody isotherm. This finding indicates that the photometric ACU method can discriminate objectively between AH and AN antibodies. (ii) The reaction of mixtures of AH and AN antibodies with homologous virus was examined and found that AH antibodies combine with virus independently from the presence of AN antibodies, whereas AH antibodies were found to reduce greatly the measurable degree of hemagglutination inhibition by AN antibodies. (iii) A formula was developed and experimentally confirmed by means of which the binding of virus by mixtures of AH and AN antibodies can be predicted. Thereby, the influence of the relative concentration of AH and AN antibodies on the establishing of isotherms and on their subsequent use for antibody titration by means of the photometric ACU method was investigated. It was found that the procedure routinely employed for establishing isotherms yielded data reflecting the reaction of AH antibodies only. As a consequence, the use of these isotherms will identify AH antibodies.

Antibodies, Viral↗
PubMed1996

Assessment of the serological relatedness of genital human papillomaviruses by hemagglutination inhibition.

To assess the potential for cross-protection among genital human papillomavirus (HPV) types in virus-like particle (VLP)-based vaccinations, inhibition of HPV VLP-mediated hemagglutination by rabbit antisera raised against HPV type 6b (HPV-6b), HPV-11, HPV-16, HPV-18, HPV-31, HPV-33, and HPV-45 was analyzed. Only highly homologous types (HPV-6b and HPV-11, and HPV-18 and HPV-45) exhibited detectable serological cross-reaction for the class of antibodies that inhibit virion-to-cell surface binding. However, analysis of neutralizing monoclonal antibodies to several animal and human papillomaviruses indicated that over half of these antibodies do not prevent cell surface binding, but these latter antibodies do not appear to be more cross-reactive in enzyme-linked immunosorbent assays than those that mediate inhibition of hemagglutination. The data strongly suggest that while there may be limited cross-protection between highly (>85% L1 amino acid identity) homologous types, protection by HPV VLP-based vaccines will be predominantly type specific.

Animals↗
PubMed1978

Comparison of rotavirus strains by hemagglutination inhibition.

Rotaviruses have been shown to be of importance as aetiologic agents of gastroenteritis in infants and in domestic animals of several species. Hemagglutinins were prepared from two Canadian isolates of bovine rotavirus and from one isolate of a simian rotavirus. A United Kingdon isolate of bovine rotavirus was shown not to possess hemagglutinating activity, indicating a strain difference between a Canadian and United Kingdom bovine rotavirus. In hemagglutination-inhibition (HAI) tests a rabbit hyperimmune (two injections) serum, prepared to one of the bovine rotaviruses, was not helpful in distinguishing the two bovine viruses because of cross-reactions between the viruses. However, it was possible to distinguish the bovine viruses from the simian virus with this serum. When guinea pig immune sera were prepared to the four rotavirus strains and tested with the three hemagglutinins in the HAI test, antigenic differences between the four strains of rotavirus were demonstrated. Hyperimmune guinea pig serum prepared to a strain of human rotavirus did not inhibit any of three hemagglutinins indicating that the human strain is different from the three rotavirus strains which gave hemagglutinins.

Animals↗
PubMed1979

Effect of serum-antigen incubation times on the expression of non-specific inhibitors of rubella hemagglutination.

Under certain conditions, serum very low-density lipoproteins (VLDL) and high-density lipoproteins (HDL) can inhibit rubella hemagglutination. The level of this non-specific hemagglutination-inhibition (HI) activity increases as the incubation period between serum and antigen is increased. Treatment of serum with heparin-MnCl2 does not precipitate HDL, and may not effect complete removal of all VLDL. This treatment method, therefore, should be considered a source of false-positive reactions, especially when extended serum-antigen incubation periods are used to enhance HI activity and to detect low levels of IgM.

Antibodies, Viral↗
PubMed1977

A reappraisal of the forces involved in hemagglutination.

The underlying principles governing the second stage of hemagglutination, as described in 1965, have been reevaluated. This reappraisal reaffirms that red cell aggregation is dependent on the potential energy barrier (zota) between erythrocytes. This in turn is dependent on the ionic strength and dielectric constant of the bulk medium. Hemagglutination by IgG and IgM antibody is a function of the effective length of these immunoglobulins and the zota of the cells in the reaction mixture. Agglutination of cells occurs when the zota-potential is below a critical level. The enhancing action of natural and synthetic polymers (e. g., bovine albumin) is to lower zeta by raising the dielectric constant of the bulk medium. Several other recent concepts are discussed and the major differences discussed in light of data available.

Antibodies, Anti-Idiotypic↗
PubMed1980

Hemagglutination antigen of Mycobacterium tuberculosis. Immunochemical characterization and concanavalin A reactivity.

The hemagglutination antigen isolated from Mycobacterium tuberculosis strain Aoyama B was characterized by immunoelectrophoresis using the International Reference System for Mycobacterial Antigens. It showed two precipitin bands, the major one being identical with antigen 1 (arabinomannan). The minor precipitin band formed a spur with antigen 2 (arabinogalactan), attesting to the presence of common determinant groups. The immunoelectrophoretic analysis of the hemagglutination antigen against concanavalin A also revealed 2 precipitin bands, the major one being identical with the single precipitin band formed between the reference antigen and concanavalin A. The 2 arabinomannans present seemed to differ in molecular weight.

Concanavalin A↗