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PubMed1980

Comparison of the hemagglutination inhibition procedure and an enzyme-linked immunosorbent assay for detection of specific antibodies to pneumonia virus of mice in experimentally infected laboratory rats.

An enzyme-linked immunosorbent assay (ELISA) and the hemagglutination inhibition (HI) test were used to evaluate the response of laboratory rats to experimental infection with pneumonia virus of mice. The ELISA procedure was more sensitive than the HI test and detected very low levels of antibodies early in the course of infection. At 5 days postinfection, ELISA detected antibody increases in five of five animals, whereas only two of five increases were detected by the HI test. At 9 days postinfection, the HI test failed to detect one titer increase measured by ELISA. Later during the course of infection, increases were detected by both tests. The ELISA procedure was, in general, more sensitive for detecting low levels of antibody than was the HI test, but was equal in sensitivity when high titers of antibody were measured.

Animals↗

PubMed1980

Comparison of passive hemagglutination, bactericidal activity, and radioimmunological methods in measuring antibody responses to Neisseria meningitidis group A capsular polysaccharide vaccine.

Passive hemagglutination (HA), a bactericidal activity test (BCA), and radioimmunoassay (RIA) were compared in measuring serum antibodies before and after group A meningococcal capsular polysaccharide vaccination of servicemen. The three methods were found satisfactory in demonstrating a response to vaccination in this age group. Of the postimmunization sera, 5% remained without HA and 1% remained without BCA activity; 1% of the postimmunization sera had less than 2 micrograms of antibody per ml as measured by RIA. Approximately 60% of the serum pairs showed a greater than or equal to 32-fold rise in HA titer, a greater than or equal 25-fold rise in BCA titer, or a greater than or equal to 4-fold rise in antibody concentration by RIA. A difference in response to two different vaccine lots was seen with RIA and BCA. Although the calculated correlation between the three methods was good, some individual sera gave discrepant results. These could be shown to be due mainly to one of the following factors: low HA titer was due to lack of the immunoglobulin M and A classes of antibodies, low BCA titer was due to the blocking effect of high immunoglobulin A content, and high BCA titer was due to antibodies directed to bacterial components other than the capsular polysaccharide.

Antibodies, Bacterial↗

PubMed1980

Passive hemagglutination technique for serotyping Campylobacter fetus subsp. jejuni on the basis of soluble heat-stable antigens.

Antigenic materials were extracted from Campylobacter fetus subsp. jejuni strains by heating bacterial suspensions in saline at 100 degrees C and by exposure to ethylenediaminetetraacetic acid. The antigens were heat stable at 100 degrees C, capable of sensitizing sheep erythrocytes for agglutination in antisera, and able to elicit production of specific antibody in rabbits; they occurred with different immunological specificities in 23 strains. Antisera against the 23 strains could be used for discriminating among isolates of the species when the passive hemagglutination technique was used for serotyping. Three serotypes were more common than others among a collection of human isolates.

Antigens, Bacterial↗

PubMed1981

Ether treatment of type B influenza virus antigen for the hemagglutination inhibition test.

Ether treatment was studied as a method of increasing the ability of type B influenza antigen to detect antibody by hemagglutination inhibition. Comparisons were made with the untreated antigen, with an eluate made from the same virus, and with a standard type B antigen of an earlier virus. Results were evaluated based on the comparative ability to detect rises in antibody titer, as well as the relative frequency of antibody prevalence determined by each method. The ether-treated antigen was far superior to the untreated antigen in both respects; it was also superior to the eluate, although the difference was less pronounced. The treated antigen performed better than the standard type B antigen in detecting antibody in children, but there was little difference in adults. This pattern was felt to be a result of the closer relation of the treated antigen to the infecting strain. The method is, therefore, proposed as a means of producing more reactive antigens of currently circulating strains of type B influenza virus.

Adolescent↗

PubMed1981

Comparison of hemagglutination inhibition test and enzyme-linked immunosorbent assay for determining antibody to rubella virus.

The hemagglutination inhibition test (HAI) and the enzyme-linked immunosorbent assay (ELISA) for detecting antibody to rubella virus were compared by testing 25 sets of paired sera taken before and after infection and 10 sets of sera taken during acute and convalescent stages of the disease and by screening 700 serum samples from the Collaborative Perinatal Project, NIH/NINCDS. The tests were found to be comparable in their ability to detect positive and negative sera, rises in titers, and seroconversions. When a purified antigen and carefully prepared reagents were used, ELISA was found to be as accurate and reliable as HAI. ELISA required no pretreatment of serum, could easily be automated, and was less time-consuming than HAI.

Antibodies, Viral↗

PubMed1981

Comparison of enzyme-linked immunosorbent assay for acute measles with hemagglutination inhibition complement fixation, and fluorescent-antibody methods.

An enzyme-linked immunosorbent assay (ELISA) using commercially available reagents was compared with standard serological tests for determination of antibody titers in patients with acute measles. The measles-specific ELISA was as sensitive as complement fixation, hemagglutination inhibition, and indirect fluorescent-antibody methods. The ELISA method used was simple, sensitive, inexpensive, and reproducible and may conveniently be used in a small clinical laboratory.

Antibodies, Viral↗

PubMed1981

Difference in antibody reactivity between complement fixation and immune adherence hemagglutination tests with virus antigens.

Complement-fixing (CF) immunoglobulin M antibody to infantile gastroenteritis virus (a rotavirus) did not show the reactivity of immune adherence hemagglutination (IAHA). Early immunoglobulin G CF antibody produced both in patients and in guinea pigs experimentally infected with Japanese encephalitis virus (a flavivirus) had weak reactivity in IAHA test. However, late antibody showed higher titers by IAHA than by CF. These results suggested that early antibodies with lower affinity are inefficient in the IAHA reaction. The implications of this study are: (i) a low ratio of IAHA/CF antibody titers in a serum suggests a recent rotavirus infection; (ii) the IAHA reaction is more type specific than the CF reaction for identifying the serotype of antigenically cross-reacting viruses with hyperimmune sera.

Animals↗

PubMed1981

Comparison of enzyme-linked immunosorbent assay and hemagglutination inhibition in a seroepidemiological study of influenza type C infection.

An enzyme-linked immunosorbent assay was developed for the detection of antibodies to influenza type C. Based on replicate testing it was decided that an increase of more than three times the standard error of the mean from one serum specimen to the next represented a significant increase in antibody content. After removal of potentially chicken egg albumin antibodies, the test appeared to be more sensitive for the detection of rises in antibody levels than was hemagglutination inhibition. In some cases, heat treatment of the serum before use in the enzyme-linked immunosorbent assay increased the readings. The strains C/AA/1/59 and C/NJ/1/76 were used in a pilot epidemiological study of influenza C infection in Tecumseh, Mich. Sera tested were collected from autumn of 1976 to autumn of 1978. The overall incidence rate for influenza C infection in the 2-year period was 7.8%. Most cases occurred in children aged 5 to 9 years, and the rates decreased rapidly with increasing age. There appeared to be no relationship to influenza C infection with influenza A or B coinfection. All 17 influenza C infections observed occurred during the 1976 to 1977 respiratory disease season. This indicates that type C virus may not occur endemically, but may exhibit year-to-year variation in infection frequency, as is the case with types A and B.

Adolescent↗

PubMed1981

Detection of rubella antibodies by hemagglutination inhibition, indirect fluorescent-antibody test, and enzyme-linked immunosorbent assay.

Using hemagglutination inhibition (HAI) as a reference method, 292 (40 nonimmune, 252 immune) human serum samples were tested by indirect fluorescent antibody (IFA) and enzyme-linked immunosorbent assay (ELISA) for immune status and quantitation of rubella antibodies. The overall agreement with HAI for immune status was 99.7% (291/292) with IFA and 98.6% (288/292) with ELISA. Two specimens (0.7%, 2/292), negative by HAI, were equivocal by ELISA. Initially a 6.5% (19/292) overall disagreement was obtained for immune status evaluation between HAI and IFA, which was reduced to 0.3% (1/292) upon repeat testing. All of these samples were near the immune/nonimmune cutoff point (95 samples), reflecting an initial disagreement of 20% (19/95) in this category (HAI titers less than 1:20). Likewise, an initial overall disagreement of 4.5% (13/292) was obtained between HAI and ELISA which was reduced to 0.7% (2/292) upon repeated testing. Eleven of the 13 samples were near the immune/nonimmune cutoff point, reflecting an initial disagreement of 11.6% (11/95) with sera having an HAI antibody titer of less than 1:20. Quantitation of rubella antibodies by IFA showed an overall correlation with HAI of 86.6% within less than twofold titer and 99.3% within less than fourfold titers. In testing the ability of ELISA to quantitate antibody, a correlation coefficient (r) of 0.996 was obtained by plotting the measured average optical density (405 nm) of ELISA against the corresponding log of HAI titer. Both IFA and ELISA showed good correlation with HAI for immune status evaluation and for quantitation of rubella antibodies. Technically the HAI was the most cumbersome to perform, whereas IFA was the least technically demanding. Originally, 308 samples were tested; 16 samples (5.2%) could not be evaluated by IFA because of a high level of nonspecific fluorescence. The strict requirement of controlling the temperature range (23 to 24 degrees C) during substrate hydrolysis proved to be a problem with the ELISA test in our laboratory.

Antibodies, Viral↗

PubMed1982

Assay of human rotavirus antibody by immune adherence hemagglutination with a cultivable human rotavirus as antigen.

A cultivable human rotavirus, Wa, was propagated in MA-104 cells and used as an antigen in immune adherence hemagglutination (IAHA) and complement fixation (CF) tests. IAHA antibody titers of sera from normal humans were found to be eightfold higher than CF antibody titers. The IAHA antibody determination with the Wa antigen is considered to be especially useful for the seroepidemiological studies of human rotavirus infections.

Antibodies, Viral↗

PubMed1982

Comparison of indirect fluorescent-antibody amoebic serology with counterimmunoelectrophoresis and indirect hemagglutination amoebic serologies.

Patients ranged from those with no prior diagnosis of or suspected exposure to Entamoeba histolytica to those with proven amoebic liver abscesses (extraintestinal disease). A comparison of serologies from patients with proven and suspected amoebiasis or possible past exposure revealed good correlation between the indirect fluorescent antibody (IFA) procedure and the other methods used, counterimmunoelectrophoresis and indirect hemagglutination. Titers from patients with proven extraintestinal amoebiasis were in the expected high range previously reported by other authors. Patients with clinical histories suggestive of exposure to E. histolytica but no proven disease had lower titers which indicated possible background exposure. The IFA procedure provides a rapid method of antibody detection; results obtained on an emergency basis provide essential information in making the diagnosis of amoebic abscess, pyogenic abscess, or tumor. The IFA procedure is rapid, reliable reproducible, and relatively inexpensive to perform, provided a good source of antigen is consistently available.

Amebiasis↗

PubMed1982

Comparative measurement of equine influenza virus antibodies in horse sera by single radial hemolysis, neutralization, and hemagglutination inhibition tests.

Single radial hemolysis (SRH), neutralization (NT), and hemagglutination inhibition (HI) tests were carried out on sera from horses immunized against the Prague and Miami strains of equine influenza virus. The HI and NT tests demonstrated good sensitivity; the sensitivity of the SRH test was somewhat lower. The NT titers of individual sera were correlated very closely with the HI titers, although the NT titers were higher. SRH zone diameters of individual sera also showed significant correlation with the NT and NI titers. The SRH test appears to be suitable for large-scale serological surveys and offers the advantages of rapidity and simplicity.

Animals↗

PubMed1982

Comparison of the latex agglutination test with the hemagglutination inhibition test, enzyme-linked immunosorbent assay, and neutralization test for detection of antibodies to rubella virus.

The ability of a rapid, latex agglutination test to diagnose rubella infection and to measure immune status was evaluated by comparison with the hemagglutination-inhibition (HAI) test, enzyme-linked immunosorbent assay (ELISA), and the neutralization (NT) test. The latex agglutination test accurately detected serological conversions in 74 pairs of sera representing 21 natural infections and 53 immunizations. The antibody levels of 276 sera from the general population were determined by latex agglutination, HAI, and ELISA. The correlation coefficients between the titers obtained by HAI and latex agglutination and by ELISA and latex agglutination were statistically significant. Results on 12 sera did not agree when measured by the three tests. These sera were included among the 196 specimens tested by NT. The correlation coefficient between NT and latex agglutination titers was statistically significant. There was one serum positive by latex agglutination but negative by NT, and five sera were negative by latex agglutination but had titers of 4 to 8 in the NT. The relative sensitivity of detecting antibody was greater by latex agglutination than by HAI. An additional 49 sera containing residual nonspecific hemagglutinin inhibitors were evaluated by latex agglutination and NT. The untreated sera showed no false positive reactions, and 36 of 39 NT positive sera were positive in the latex agglutination test.

Antibodies, Viral↗

PubMed1983

Fluorescence immunoassay and passive latex agglutination as alternatives to hemagglutination inhibition for determining rubella immune status.

Three different assays for detection of rubella antibodies, hemagglutination inhibition (HAI), fluorescence immunoassay (FIA), and passive latex agglutination (PLA) were used to test 297 human serum samples. Overall agreements for immune status were as follows: HAI versus FIA, 95.3% (283 of 297); HAI versus PLA (1:10 dilution), 96.3% (286 of 297); HAI versus PLA (undiluted), 93.9% (279 of 297); PLA (1:10 dilution) versus FIA, 94.9% (282 of 297); and PLA (undiluted) versus FIA, 97.9% (291 of 297). The HAI test is the most time consuming, subjective, and technically difficult to perform. The FIA and PLA tests are very rapid and less labor intensive. In addition, the FIA offers an objective determination of the patient's rubella antibody level.

Antibody Specificity↗

PubMed1983

Microtiter determination of measles hemagglutination inhibition antibody with filter papers.

Epidemiological studies of measles and measles immunization frequently require determination of measles antibody status. In developing countries, where venipuncture is frequently unacceptable and where refrigerated storage of serum specimens is often unavailable, microtiter techniques not requiring refrigeration are required. We developed a filter paper technique that measures measles hemagglutination inhibition antibody and meets these criteria. Comparison of separately collected venous blood and peripheral blood collected on filter paper demonstrated 97% agreement in terms of presence or absence of antibody. In 30 of 32 measles specimens, 94% of titers were the same or varied by less than 2 twofold dilutions.

Antibodies, Viral↗

PubMed1983

Rapid indirect hemagglutination test for serotyping Pasteurella haemolytica.

A modified indirect hemagglutination test which enabled Pasteurella haemolytica to be serotyped in 3 h is described. There was 100% correlation with the conventional test for both the 15 prototype strains and the 50 field isolates. It was found that nonviable cultures could also be serotyped.

Hemagglutination Tests↗

PubMed1983

Hemagglutination treponemal test for syphilis.

Sera from 290 hospital patients were tested to compare the sensitivity, specificity, and reproducibility of the hemagglutination treponemal test for syphilis (HATTS) with the fluorescent treponemal antibody absorption test (FTA-ABS). Complete agreement was obtained between the methods when 142 syphilitic sera from patients with various stages of syphilis were tested. By using clinical histories, the specificity with 148 nonsyphilitic sera was determined to be 100% for the HATTS and 96.6% (143 of 148) for the FTA-ABS. Satisfactory reproducibility was obtained with both methods. Compared with the FTA-ABS, the HATTS was more specific, easier, and more economical to perform. We therefore recommend the HATTS as a suitable alternative to the FTA-ABS.

Cost-Benefit Analysis↗

PubMed1983

Increased sensitivity and reduced specificity of hemagglutination inhibition tests with ether-treated influenza B/Singapore/222/79.

Hemagglutination inhibition (HI) tests against whole virus (WV) influenza B/Singapore/222/79 antigen detected prevaccination serum antibody in only 15 (20%) of 50 predominantly elderly volunteers and fourfold or greater titer rises in only three (6%) after they received 1981-1982 trivalent influenza vaccine containing antigens of this virus. HI titers against ether-treated (ET) B/Singapore/222/79 were about eightfold higher than those against WV antigen and were comparable to microneutralization titers against this virus. The ET HI detected prevaccination antibody in 84%, a postvaccination titer rise in 32%, and a final titer of 80 or higher in 66%. Among 51 additional persons with known or presumed influenza B virus infections early in 1982, ET B/Singapore/222/79 was also more sensitive than WV for serodiagnosis (69 versus 49%), but eight persons with both WV and ET B/Singapore/222/79 HI responses also had an HI titer rise to WV A/Brazil/11/78 (H1N1) antigen. Conversely, among 14 college students with febrile, culture-proven influenza A (H1N1) infections early in 1982, 6 (43%) developed HI titer rises to ET B/Singapore/222/79 with no other serological evidence of influenza B virus infection. Moreover, young adult volunteers with mild experimental influenza A (H1N1) infections also exhibited a 17% (3 of 18) incidence of ET B/Singapore/222/79 HI titer rises, versus none in matched, uninfected volunteers. These data indicate that ET B/Singapore/222/79 virus has increased sensitivity but reduced specificity compared to WV as an HI antigen and that caution is needed in interpretation of a single HI test for serodiagnosis, whether with WV or ET antigen.

Aged↗