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PubMed1979

Hemagglutination by Escherichia coli in septicemia and urinary tract infections.

The agglutination of erythrocytes from various animal species by Escherichia coli was studied. The 405 strains of E. coli were isolated from urine in patients with urinary tract infections, from blood in septicemic patients, or from feces in persons without intestinal or urinary disorders. In urinary tract infections, d-mannose-resistant agglutination (MRHA) of human erythrocytes was the most common finding (23% of the strains). The highest frequency of mannose-sensitive hemagglutination (MSHA) attributed to type I (common type) pili occurred with guinea pig erythrocytes (11.5%). Of the 78 E. coli strains isolated from blood cultures, 11 (14%) produced MRHA of human erythrocytes and only one gave MSHA. In the stool cultures, only 1 of 170 E. coli strains was MSHA reacting, whereas 28 strains (16.5%) showed MRHA of human erythrocytes. No MRHA strain reacted with antiserum against colonization factor antigen (CFA)/I of pilus nature in enterotoxigenic human E. coli strains (O78:H12). MRHA of bovine erythrocytes, reputedly typical of enterotoxigenic E. coli of serogroups O6 and O8, was shown by only two strains, neither of which agglutinated with CFA/II antiserum. The most common hemagglutinating pattern of E. coli from urine and blood thus was MRHA for human erythrocytes. This agglutination may have been caused by pili or other surface properties of one or more serotypes. These may represent a new class of colonization-promoting antigens (adhesins).

Animals↗
PubMed1981

Comparison of enzyme-linked immunosorbent assay and hemagglutination test using mycobacterial glycolipids.

Enzyme-linked immunosorbent assay and hemagglutination methods were compared using mycobacterial glycolipids as antigens. Both methods were found to have equivalent specificity and sensitivity in detecting mycobacterial diseases. Both tests had 96% specificity; the sensitivity of the enzyme-linked immunosorbent assay was 86%, and that of the hemagglutination test was 88.6%.

Antibodies, Bacterial↗
PubMed1981

Antibodies to the enterobacterial common antigen: standardization of the passive hemagglutination test and levels in normal human sera.

The passive hemagglutination test for antibodies against the enterobacterial common antigen (ECA) of Kunin was standardized for diagnostic purposes. Human erythrocytes were coated with a soluble ECA+ preparation from Salmonella typhimurium or, as specificity controls, with a similar ECA- preparation from congenic ECA-negative bacteria with saline, and the hemagglutination assay was performed on microtiter plates. The specificity of the test was ascertained further by inhibition assays with purified ECA and with crude ECA+ and ECA- preparations. The reproducibility of the tests was 96.4%; on this basis, a fourfold or larger difference in titers was regarded as significant. The anti-ECA titers in 649 serum samples from healthy persons ranged from less than 4 to 8,192. The titers increased with age, so that th geometric mean titers were 57 at 1.5 years of age and 201 at 45 years of age. After this, the titers decreased again, to a low of 52 in persons more than 70 years old. Women had higher titers than men up to the age of 40 years.

Adolescent↗
PubMed1981

Specificity, sensitivity, and reproducibility among the fluorescent treponemal antibody-absorption test, the microhemagglutination assay for Treponema pallidum antibodies, and the hemagglutination treponemal test for syphilis.

Using 920 sera, we compared the specificity and reproducibility of the hemagglutination treponemal test for syphilis with those of the fluorescent treponemal antibody-absorption test and the microhemagglutination assay for Treponema pallidum antibodies; we found all three tests to be comparable. However, the hemagglutination treponemal test for syphilis, like the microhemagglutination assay for T. pallidum antibodies, lacked sensitivity in sera from patients with primary syphilis.

Antibodies, Bacterial↗
PubMed1982

Passive hemagglutination test for detection of antibody to gonococcal ribosomal antigen in sera from patients with asymptomatic gonorrhea.

Ribosomal fractions were obtained from a culture of type 2 Neisseria gonorrhoeae strain P-17 which was isolated from a patient with an acute gonococcal infection; these fractions were purified to eliminate the components of the outer membrane complex by affinity chromatography (Sepharose-anti-outer membrane complex antibody conjugates were used as the solid immunosorbent), and the resulting preparation was designated the purified ribosomal fraction, The purified ribosomal fraction was used to detect antibody activity in sera obtained from culture-positive asymptomatic carriers and healthy controls by a passive hemagglutination test. This passive hemagglutination test had a specificity of 100% for both sexes and sensitivities of 99.4 and 88.2% for female and male carriers, respectively, when an antibody titer of more than 1:3 was defined as abnormal. Absorption of the sera with nongonococcal organisms did not affect the antibody activity, and no significant difference in antigenicity among various N. gonorrhoeae strains was observed in ribosomal fractions. An enzyme-linked immunosorbent assay was also used to measure the relative amounts of specific antibodies to the purified ribosomal fraction, and this assay revealed that the anti-purified ribosomal fraction antibodies were immunoglobulin G.

Antibodies, Bacterial↗
PubMed1982

Indirect hemagglutination test that uses glutaraldehyde-fixed sheep erythrocytes sensitized with extract antigens for detection of Pasteurella antibody.

Glutaraldehyde-fixed sheep erythrocytes (GA-SRBC) were used in the indirect hemagglutination test for the detection of Pasteurella antibody. GA-SRBC were stable for at least 6 months. Heat extract or potassium thiocyanate extract antigens of Pasteurella strains could be adsorbed onto GA-SRBC or tanned GA-SRBC, respectively. The indirect hemagglutination test reaction was capsular group specific with heat extract antigen-sensitized GA-SRBC but not potassium thiocyanate extract antigen-sensitized tanned GA-SRBC.

Animals↗
PubMed1983

Determination of antigenic specificity and relationship among Haemophilus pleuropneumoniae serotypes by an indirect hemagglutination test.

Serological properties of antigens extracted from strains of Haemophilus pleuropneumoniae belonging to seven different serotypes were investigated. Antisera were prepared in rabbits against Formalin-treated whole cell suspensions as well as autoclaved cell suspensions. Saline and heat extracts and their alcohol precipitate antigens of H. pleuropneumoniae were used in the indirect hemagglutination test. All the antigens used were easily adsorbed directly onto sheep erythrocytes. Saline extract antigen showed maximum type specificity. Heating of the whole cell suspension revealed the cross-reactive minor antigenic determinants. Thus, the heat extract preparations had both type-specific and species-specific antigens. It is suggested that the indirect hemagglutination test may be useful for both serotyping and serodiagnosis of H. pleuropneumoniae infections in pigs.

Antigens, Bacterial↗
PubMed1983

Correlation of an expanded direct fluorescent-antibody system with an established passive hemagglutination system for serogrouping strains of Campylobacter jejuni and Campylobacter coli.

Rabbits were inoculated with whole, formalinized cells from eight passive hemagglutination reference strains of Campylobacter. Fluorescein isothiocyanate-labeled immunoglobulin G from these antisera defined seven new direct fluorescent-antibody serogroups of C. jejuni and one new serogroup of C. coli. This expanded the Campylobacter direct fluorescent antibody system to include 17 serogroups of C. jejuni, 3 serogroups of C. coli, and 2 serogroups of C. fetus. We then compared the passive hemagglutination method (57 serotypes) and the direct fluorescent-antibody method (20 serogroups) for typing strains of C. jejuni and C. coli. The data obtained by testing 101 strains by both methods revealed that the two test systems were measuring completely different sets of antigen complexes. The two serogrouping methods were complementary, and their combined use discriminated among strains more effectively than did either method individually.

Campylobacter↗
PubMed1983

Rubella antibodies detected by several commercial immunoassays in hemagglutination inhibition-negative sera.

Although a very good correlation was found between the level of rubella antibodies measured by a standard hemagglutination inhibition (HI) test and by an enzyme-linked immunosorbent assay (ELISA) procedure (Cordia R), an appreciable proportion (31%) of ELISA-positive specimens were encountered among HI-negative sera. The reverse was rarely seen. Many of the HI-negative, ELISA-positive sera were also found to be positive for rubella antibodies by one or more other assay methods, including an immunofluorescence assay (IFA) procedure (FIAX), passive hemagglutination (PHA) (Rubacell and PHAST), latex agglutination (Rubascan), and a second ELISA procedure (Rubelisa). The specificity of all of the ELISA-positive HI-negative specimens was substantiated by absorption experiments. In these tests, the ELISA reactivities were blocked by rubella antigens, but not by a variety of tissue culture control antigens or by influenza virus grown on the same cell line. The findings indicate that many of the newer methods available for rubella antibody detection are more sensitive than HI for detecting low levels of rubella antibodies. Until more clinical information is available concerning the protective nature of these low levels of antibody, caution should be exercised in assessing the significance of these results.

Antibodies, Viral↗
PubMed1984

Immunity to human cytomegalovirus measured and compared by complement fixation, indirect fluorescent-antibody, indirect hemagglutination, and enzyme-linked immunosorbent assays.

The complement fixation test is currently the test employed most frequently to determine the presence of antibody to human cytomegalovirus. Several other techniques have been adapted for this purpose. A comparison of cytomegalovirus antibody titers was made between the complement fixation test, a commercially available enzyme-linked immunosorbent assay, an indirect immunofluorescent technique, and a modified indirect hemagglutination test. Forty-three serum samples were tested for antibodies by each of the above procedures. The enzyme-linked immunosorbent, immunofluorescent, and indirect hemagglutination assays were in close agreement on all samples tested; the titers obtained with these methods were all equal to or greater than the complement fixation titer for 38 of the 41 samples (92.6%). Two samples were anticomplementary in the complement fixation test but gave readable results in the other tests. The complement fixation test was the least sensitive of the procedures examined. The commercial enzyme-linked immunosorbent assay system was the most practical method and offered the highest degree of sensitivity in detecting antibodies to cytomegalovirus.

Antibodies, Viral↗
PubMed1984

Quantitation of enterovirus 70 antibody by microneutralization test and comparison with standard neutralization, hemagglutination inhibition, and complement fixation tests with different virus strains.

We describe here a microneutralization procedure for conveniently testing large numbers of specimens for antibodies to enterovirus 70. The test utilized human rhabdomyosarcoma cells and was read by staining with crystal violet after 4 days of incubation. The test compares well with other serological assays, being more sensitive than the standard tube neutralization test and the complement fixation test, but less sensitive than the hemagglutination inhibition test. However, the hemagglutination inhibition test required concentrated, partially purified virus as antigen, as did the complement fixation test, and was difficult to read, so that its greater sensitivity may not be of practical significance. By all four test procedures, a recent isolate of enterovirus 70 was a more sensitive antigen than the prototype strain, as shown by greater geometric mean titers in sera of patients from various epidemics.

Antibodies, Viral↗
PubMed1984

Detection of cytomegalovirus antibody with two commercially available assays, an indirect hemagglutination test and an enzyme immunosorbent assay.

By the use of two reference procedures, an indirect hemagglutination assay and a complement fixation test, the presence or absence of cytomegalovirus (CMV) antibody was determined for 221 human sera. Ninety-nine sera (44.8%) were found to contain CMV antibody. The remaining 122 sera (55.2%) lacked detectable CMV antibody. These same sera were then analyzed by two recently introduced, commercially available CMV antibody assays, an indirect hemagglutination test (IHA-c; Cetus Corp., Emeryville, Calif.) and an enzyme-linked immunosorbent assay (ELISA; M. A. Bioproducts, Walkersville, Md.). With the results of the reference procedures as true evidence of the presence or absence of CMV antibody, the sensitivity of the IHA-c was found to be 100%; the specificity was 98.4%. The sensitivity of the ELISA was also 100%; the specificity was 96.7%. The overall accuracies of these procedures were 99.1 and 98.2%, respectively. Time and motion studies revealed the IHA-c procedure to be faster and technically less demanding than the ELISA procedure.

Antibodies, Viral↗
PubMed1984

Comparison of enzyme-linked immunosorbent and indirect hemagglutination assays for determining anthrax antibodies.

An enzyme-linked immunosorbent assay has been established to measure anthrax antibody titers. The protective antigen component of anthrax toxin was used as the capture antigen. Two types of conjugates (protein A-horseradish peroxidase and anti-human immunoglobulin G plus immunoglobulin A plus immunoglobulin M-horseradish peroxidase) were tested. Results from enzyme-linked immunosorbent assay testing were compared with those from indirect hemagglutination titers on serum from vaccinees. The overall trend of enzyme-linked immunosorbent assay and indirect hemagglutination titers was significant. The enzyme-linked immunosorbent assay offered speed, precision, and reduced cost per test.

Antibodies, Bacterial↗
PubMed1985

Improvement of the indirect hemagglutination assay for Salmonella typhi Vi antibodies by use of glutaraldehyde-fixed erythrocytes.

Glutaraldehyde-fixed sheep erythrocytes sensitized with Vi antigen were shown to be usable for at least 1 year in the indirect hemagglutination assay for Vi antibodies. The use of fixed sensitized erythrocytes improves the practicality of the indirect hemagglutination assay by (i) eliminating the need to sensitize cells each time the test is performed; (ii) reducing waste of the purified Vi antigen; and (iii) reducing test-to-test variation.

Animals↗
PubMed1986

Confirmation of human cytomegalovirus by reverse passive hemagglutination with monoclonal antibodies reactive to the major glycosylated peptide (GP-66).

Sheep erythrocytes coated with three monoclonal antibodies, each reactive to a different epitope of the 66-kilodalton cytomegalovirus (CMV) matrix protein, were used in a reverse passive hemagglutination test with CMV-infected cell lysate to identify and confirm the CMV. The test is specific only for CMV, since 5 laboratory strains of CMV (AD169, Davis, Espilat, C-87, and Towne) and 10 clinical isolates reacted well, but uninfected MRC-5 cell lysate, lysates of herpes simplex virus types 1 and 2, varicella-zoster virus, and adenoviruses did not react. The reactive CMV lysate was confirmed by the pretreatment of CMV lysate with the three monoclonal antibodies followed by the addition of antibody-coated erythrocytes. The reverse passive hemagglutination test and the confirmatory blocking test are performed at the same time, requiring 2 h to complete. Since V-bottom microtiter 96-well plates and a 25-microliter pipette can be used to perform the test, it is ideal for CMV confirmation, especially when the equipment to read the fluorescent-antibody test or enzyme-linked immunosorbent assay is not available.

Antibodies, Monoclonal↗
PubMed1986

Passive hemagglutination test for antibody to hepatitis B core antigen.

Available tests for antibody to hepatitis B core antigen (anti-HBc) are complicated. A passive hemagglutination assay for anti-HBc was developed by sensitizing sheep erythrocytes with hepatitis B core antigen synthesized by a recombinant DNA technique. It was compared with a commercial passive hemagglutination assay and an enzyme-linked immunosorbent assay for anti-HBc and showed good agreement with both. It is rapid, simple, and sensitive.

DNA, Recombinant↗
PubMed1987

Monoclonal antibody passive hemagglutination and capture enzyme-linked immunosorbent assays for direct detection and quantitation of F41 and K99 fimbrial antigens in enterotoxigenic Escherichia coli.

Production of diarrhea in neonatal calves by enterotoxigenic Escherichia coli depends on its ability to attach to the epithelial cells of the intestine via surface adhesins called pili or fimbriae and to secrete enterotoxins. The most important of these fimbriae are designated K99 and F41. We produced and characterized a murine monoclonal antibody specific to F41. This monoclonal antibody and a K99-specific monoclonal antibody were used to develop sensitive and specific passive hemagglutination and capture enzyme-linked immunosorbent assays (ELISAs) for detection and quantitation of F41 and K99 antigens in E. coli cultures and culture supernatants. The capture ELISA systems exhibited excellent sensitivity and specificity, whereas the passive hemagglutination systems appeared to be oversensitive. The ability of the capture ELISAs to detect K99 and F41 fimbrial antigens in fecal specimens from calves was evaluated. Fimbrial antigens were detected in six of six specimens from scouring calves but not in four of four specimens from nonscouring calves.

Animals↗