[A new indirect hemagglutination test in the determination of anti-toxoplasma antibodies].
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The indirect haemagglutination test has been developed for use with LCM and Tacaribe viruses employing antibody-sensitized erythrocytes. Both in this test and in the indirect haemagglutination inhibition test cross-reactions between LCM and Tacaribe viruses were revealed. This opens new perspectives for group and species identification of arenaviruses.
Haemagglutination inhibition
Arenavirus
Hemagglutination tests
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The influence of various concentrations of human γ-globulin (HGG) employed to sensitize tanned sheep erythrocytes on haemagglutination titres with various early and late mouse and rabbit antisera to HGG has been studied. The concentration of HGG employed to sensitize tanned erythrocytes which gives the highest haemagglutination titres with early, mercaptoethanol-sensitive antibodies was not optimal for obtaining highest haemagglutination titres with late, mercaptoethanol-insensitive antibodies. Similar results were obtained with heavy and light sucrose gradient ultracentrifugation fractions of a late and early rabbit antiserum. These findings may account for past discrepancies and should be considered when employing the haemagglutination test to detect early and late antibodies.
Haemagglutination inhibition
Hemagglutination tests
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In this paper, the results are reported of experiments to investigate the optimum conditions for determining, and differentiating between, the antibodies against closely related poxviruses using the ELISA technique. This assay was found to be considerably more sensitive than the virus neutralization, haemagglutination inhibition, passive haemagglutination, and indirect fluorescent antibody tests for detecting antibodies, but has not yet been compared with radioimmunoassay. The use of ELISA made it possible to differentiate between antibodies to vaccinia, whitepox, and monkeypox viruses.
Monkeypox
Poxviridae
Haemagglutination inhibition
Orthopoxvirus
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Twenty strains of Venezuelan encephalitis (VE) virus inoculated intravenously in large doses into roosters produced hemagglutination-inhibition (HI) antibodies detectable in plasmas within 7 to 10 days. No signs of illness occurred, and there was no evidence of viral growth in tissues since blood concentrations of infectious virus steadily decreased after inoculation. HI antibodies in early plasmas were specific for VE virus and did not cross-react significantly with two other North American alphaviruses, eastern and western encephalitis viruses. VE virus strains could be distinquished by virus-dilution, short-incubation HI, but not by plasma-dilution neutralization tests, by using early rooster antibodies. The distinctions by HI test were similar with some strains to, but different with other strains from, those described by Young and Johnson with the spiny rat antisera used to establish their subtype classifications of VE virus (14, 28). Nevertheless, results of HI tests with rooster antibodies correlated with equine virulence, as did results with spiny rat antibodies, and distinguished the new strains of virus that appeared in Middle America during the VE outbreak of 1969 from preexisting strains.
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New england
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Summary Erythrocytes from various species showed a gradient of sensitivity to agglutination by arboviruses, from reptile erythrocytes, relatively insensitive, to avian erythrocytes, the most sensitive tested. Lipids extracted from both inagglutinable and agglutinable erythrocytes inhibited arbovirus haemagglutination. Lipid fractionated on DEAE-cellulose columns yielded two major inhibitory fractions, one containing only neutral lipid including cholesterol and another phospholipid and neutral lipid but no cholesterol. Fractionation of the lipid on silicic-acid columns resulted in a loss of activity in the fractions compared with the total lipid extract. Recombination of the fractions did not restore the inhibitory activity. Similar loss of activity followed treatment of lipid extracts with digitonin. No evidence of an essential component in inhibitory lipid mixtures was observed. Tests of mixtures of standard lipids gave results suggesting the need for suitable arrangements of lipids in aqueous solutions for interaction with arboviruses. Only certain combinations of lipids were potent inhibitors. The interactions observed were thought to be between arboviruses, which contain lipid, and artificial lipid aggregates in aqueous solutions. The arrangements of lipids were not thought analogous to receptor sites on erythrocytes.
Agglutination (biology)
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When commercial erythrocytic diagnostic agents, prepared from sheep red cells, are used in the passive hemagglutination test with blood sera from patients with intestinal infections, the possibility of nonspecific reactions with heterophilic antibodies of human body should be borne in mind. To completely eliminate these antibody effects on the results of the test, titration of the tested serum should be started from 1:64, 1:80, or 1:100, depending on what titration scale is used.
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A serological surveillance was carried out to detect antibody against influenza A virus in chicken sera. A total of 8,850 field samples were collected from 47 prefectures in Japan. Initially, all the sera were screened by agar gel immunodiffusion and those sera showing positive reaction were investigated for haemagglutination-inhibition (HI) and neuraminidase-inhibition antibodies against influenza viruses. Only 6 samples had antibodies; 4 sera had antibodies against human subtype H1N1 virus; with HI activity against strain A/PR/34; three sera had strong HI activity to strain A/Tottori/4/87, which by haemagglutination test is closely related to A/Yamagata/120/86. The remaining two chicken sera had antibodies against avian subtypes H10N4 and H3N6 viruses respectively.
Haemagglutination inhibition
Immunodiffusion
Ouchterlony double immunodiffusion
Strain (injury)
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Formalinized goose erythrocytes were used in haemagglutination inhibition (HI) and indirect fluorescent-antibody (IFA) tests to detect antibodies to Japanese encephalitis (JE) and West Nile (WN) viruses in equines. Paired serum samples from 31 cases having clinical symptoms of flaviviral infections (JE and WN viruses) and 45 controls were examined. For HI test, formalinized goose erythrocytes were used as such, whereas in IFA test, formalinized goose erythrocytes were first coated with respective viral antigens separately and later used to detect antibodies. By employing HI and IFA tests, paired samples having a titre same or less than two fold rise over the control sera were considered normal for both the viruses. IFA test was found to be a method of choice, due to its sensitivity over HI test.
Flavivirus
Haemagglutination inhibition
Direct fluorescent antibody
Hemagglutination assay
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Avian influenza viruses replicate in a variety of mammals and birds, yet hemagglutination inhibition tests show that postinfection sera from these animals (e.g., ferrets and ducks) have insignificant levels of antibodies (Hinshaw et al., Infect. Immun. 34:354-361, 1981). This suggested that avian influenza viruses, in contrast to mammalian viruses, may not induce a significant humoral response. Studies reported here indicate that avian influenza viruses do induce high levels of antibodies in ferrets, ducks, and mice and produce long-lived memory for cytotoxic T-cells in mice. The failure to detect hemagglutination-inhibiting antibodies to avian viruses was explained by the finding that antibodies to avian influenza viruses were not detectable in hemagglutination inhibition tests with intact virus yet were readily demonstrable when hemagglutinin subunits were used. In addition, these sera contained high levels of neutralizing antibodies to the avian virus. These findings suggest that the hemagglutinins of avian and mammalian influenza viruses may differ in their accessibility to antibodies or the biological consequence of antibody attachment or both. The practical consequence of these studies is that hemagglutination inhibition tests with intact avian viruses fail to detect antibody and do not correlate with virus neutralization. The avian virus used in these studies, A/Mallard/NY/6870/78 (H2N2), replicated and caused mortality in BALB/c mice, emphasizing that the host range and virulence of avian viruses extends to mammals. The above findings suggest that avian viruses could infect mammals in nature, yet seroepidemiological studies with conventional hemagglutination inhibition tests could give misleading results.
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