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Hemagglutination-inhibition tests with different strains of equine infectious anemia virus.

The serologic relationships between 6 strains of equine infectious anemia (EIA) viruses were investigated by hemagglutination-inhibition (HI) tests. Cross HI tests, using sera from horses in the early stage of infection, revealed that all strains were inhibited only by homologous strain antisera and that HI antibody was always detectable before virus-neutralizing antibody. In the later stages of infection, both homologous and heterologous HI antibodies were detected in a sera of most of the horses, and the order of appearance of heterologous HI antibodies was random in 2 horses inoculated with the same virus strain. The viruses sequentially isolated from 1 horse had serologically the same hemagglutinating antigenicity to their parent virus. These results indicate that hemagglutinating antigens of EIA virus were type specific, but different from the antigens detectable by neutralization. The possible role of EIA virus hemagglutinin and HI antibody in causing periodic anemia is discussed.

Animals↗

Measurement of antibodies to avian influenza virus A(H7N7) in humans by hemagglutination inhibition test.

During the epizootic of highly pathogenic avian influenza A(H7N7) in 2003 in The Netherlands, RT-PCR and culture confirmed infection was detected in 89 persons who were ill. A modified hemagglutination inhibition (HI) test using horse erythrocytes and 2 hemagglutinating units of virus was applied to assess retrospectively the extent of human (subclinical) infection. Validation of the HI-test with sera from 34 RT-PCR and culture confirmed A(H7) infected persons and sera from 100 persons from a human influenza vaccine trial in autumn 2002 showed that this HI-test had a sensitivity of 85% and a specificity of 100% when using a cut-off titer of > or =10. Using this cut-off value, A(H7) specific antibodies were detected in 49% of 508 persons exposed to poultry and in 64% of 63 persons exposed to A(H7) infected persons. Correlation of seropositivity with the occurrence of eye symptoms in exposed persons who had not received antiviral prophylaxis and of reduced seropositivity with taking antiviral prophylaxis provided further evidence that the A(H7) HI antibody titers were real. In conclusion, by applying an HI-test using horse erythrocytes human antibodies against the avian A(H7N7) virus were detected with high sensitivity and specificity in an unexpectedly high proportion of exposed persons.

Adolescent↗

Passive hemagglutination-inhibition test for typing foot-and-mouth disease virus.

In addition to currently used serological tests for the occurrence of foot-and-mouth disease virus (FMDV), a specific "passive" hemagglutination-inhibition (HAI) test has been developed as a supplement. Serial twofold dilutions of antiserum (0.05 ml) were mixed with 0.05 ml of a constant concentration of FMDV. After incubating for 30 min at 37 C, agglutinating antibodies were determined by adding 0.1 ml of 2.5% virus-sensitized erythrocytes. The minimum concentration of antiserum required to agglutinate the erythrocytes defined the inhibition in the HAI test. Similar tests using different concentrations of virus to inhibit antibodies were carried out in parallel fashion. The relationship between the logarithm of the HAI titer and the concentration of inhibiting virus was nearly first order (P > 0.25). The slope was used as a measure of the relative specificities of the antigen-antibody interaction and was independent of concentration. The HAI test was type-, subtype-, strain-, and variant-specific with the viral antigens used. In particular, typing was performed directly on bovine antisera.

Animals↗

Enzyme immunoassay, complement fixation and hemagglutination inhibition tests in the diagnosis of influenza A and B virus infections. Purified hemagglutinin in subtype-specific diagnosis.

The efficacy of enzyme immunoassay (EIA) in detecting diagnostic antibody rises to influenza A and B viruses was compared with complement fixation (CF) and hemagglutination inhibition (HI) tests in 455 patients with an acute respiratory infection. EIA and HI detected significantly more diagnostic antibody rises against influenza A than the CF method (96 and 87 vs. 47, respectively). In the case of influenza B significantly more diagnostic influenza B antibody rises were observed by EIA than by CF or HI (59 vs. 37 and 40, respectively). In most of the cases antibody rises in EIA were found in both IgG and IgA isotypes whereas increases in IgM antibodies were seen less frequently. Purified hemagglutinins (HA) were prepared from influenza A HI- and H3-subtypes and from influenza B viruses and used as antigens in EIA and the results were compared with those of HI. Infections caused by influenza A HI-subtype showed good homologous antibody responses in EIA but heterologous antibody responses to H3-subtype and influenza B HAs were frequently observed. Heterologous responses were clearly less frequent in patients with infections caused by the H3-subtype. Influenza B infections occasionally raised HA antibodies against influenza A H1-subtype but not to the H3-subtype. Interestingly, HI detected these heterologous responses at least as frequently as EIA. When whole viruses were used as antigens in EIA, subtype specificity was not observed and cross-reactions between influenza A and B virus antibodies were found. These observations suggest that, although EIA can show greater diagnostic efficacy over HI and CF methods, HI is still the serological method of choice in determining the causative subtype of influenza A virus infection.

Adolescent↗

Comparison of Mycoplasma synoviae hemagglutinating antigens by the hemagglutination-inhibition test.

Mycoplasma synoviae (MS) strains were isolated from the trachea of hens from MS-positive broiler breeder flocks having progeny condemnations due to airsacculitis. Hemagglutinating (HA) antigens were made from several strains. The HA antigen made from the 95th medium passage of MS FMT strain was compared with that of the standard MS WVU 1853 strain by the hemagglutination-inhibition (HI) test. Sixty-six sera from 10 MS-positive flocks, 4 Mycoplasma gallisepticum (MG)-positive sera, and 7 normal sera were used in the HI test. The geometric mean (GM) HI titer with MS WVU 1853 antigen was 98.69 on 66 sera from 10 MS-positive flocks, whereas the GM HI titer with MS FMT antigen was 226.21.

Animals↗

Serotyping of Haemophilus paragallinarum by the Page scheme: comparison of the use of agglutination and hemagglutination-inhibition tests.

Seventy-two isolates of Haemophilus paragallinarum were serotyped according to the Page scheme, using a new hemagglutination-inhibition (HI) test. The results were compared with the plate agglutination method conventionally used in the Page scheme. The HI test used washed cells of H. paragallinarum, glutaraldehyde-fixed chicken erythrocytes, and rabbit antisera originally produced for the agglutination method. For 49 of the isolates, there was complete correlation between the results of the HI serotyping test and the previously performed agglutination test--23 were serovar A, two were serovar B, and 24 were serovar C. The other 23 isolates were nontypable by the agglutination test, but 21 of them could be serotyped by the HI method--six as serovar A, two as serovar B, and 13 as serovar C. Nine isolates required treatment of the bacterial cells with hyaluronidase for the expression of hemagglutination (HA) activity. Two isolates did not have HA activity despite hyaluronidase treatment and so could not be serotyped by the HI test.

Agglutination Tests↗