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REOVIRUS HEMAGGLUTINATION: INHIBITION BY N-ACETYL-D-GLUCOSAMINE.

Purified reoviruses types 1, 2, and 3 were specifically inhibited in their hemagglutination of human erythrocytes by 250 micromoles or more of N-acetyl-D-glucosamine but not by over 20 other sugars tested. N-acetyl-D-glucosamine inhibited reovirus hemaglutination by binding to capsid virus; it did not attach to the erythrocytes. It is possible that reovirus hemagglutination involves union between N-acetyl-D-glucosamine on the surface of the red cell and the glycoprotein of the virus coat.

Acetylglucosamine↗

Hemagglutination assay for antigen and antibody associated with viral hepatitis.

Hemagglutination assays are described for measuring hepatitis-associated Australia antigen and antibody. Red cells coated with isolated antigen, with chromic chloride as a coupling agent, are used for detection of antibodies. Detection of the antigen in serums depends on inhibition of hemagglutination. The test has the sensitivity and rapidity of the best tests available, is simpler to perform, and lends itself to large-scale screening of blood donors.

Antibodies↗

Microtiter indirect hemagglutination procedure for identification of streptococcal M-protein antibodies.

A number of investigators have attempted to utilize the hemagglutination system for detection of streptococcal type-specific antibody in human sera. Cross-reactions have made the procedure unreliable without cumbersome and time-consuming manipulation of the test sera. A method is described in which a microtiter indirect hemagglutination technique, using sensitized sheep erythrocytes, is sensitive, specific, and reliable for titration of type-specific antibody after naturally acquired or induced streptococcal infection.

Animals↗

Hemagglutinating property of Haemophilus aegyptius.

Extracts possessing the capacity to hemagglutinate normal human erythrocytes were recovered from Haemophilus aegyptius by treatment with either diethylene glycol or acetone. Antisera prepared against these extracts or the unextracted bacterial cell inhibited hemagglutination by homologous and heterologous antigens. Microgel diffusions indicated the presence of identical components in each extract as expressed by lines of identity between antisera to each fraction. The hemagglutinin was identified as a lipopolysaccharide, 42% lipid and 57% carbohydrate. The determination of 6% phosphorus in the lipid fraction identified it as containing phospholipid.

Antigens↗

Preparation of La Crosse virus hemagglutinating antigen in BHK-21 suspension cell cultures.

Hemagglutinating and complement-fixing antigens of La Crosse virus (California arbovirus group) were produced in serum-free suspension cultures of BHK-21/13S cells. The appearance and production of these antigens were correlated with the titer of infectious virus. No significant differences in antigen titers were produced by varying virus dose 10-fold. Hemagglutinin appeared 6 to 8 hr after inoculation and reached peak titer in 14 to 22 hr. Both beta-propiolactone and Tween 80-ether treatment inactivated infectious virus in the antigens. Unlyophilized antigen was stable at -60, 5 and 24 C for at least 117 days but not for 1 year. Lyophilized antigen was stable for at least a year, however, at -20 and 5 C. Cell culture-produced antigen was more sensitive than brain-produced antigen in detecting hemagglutination inhibition antibody in human sera.

Animals↗

Variables of the rubella hemagglutination-inhibition test system and their effect on antigen and antibody titers.

A systematic study was made of certain variables of the rubella hemagglutination-inhibition (HI) test system and their effect on antigen and antibody titers. Erythrocytes from pigeons and 1-day-old chicks gave similar antigen and antibody titers, but goose erythrocytes gave lower titers. Indicator erythrocytes could be stored in Alsever's solution at 4 C for as long as 2 weeks without losing sensitivity in hemagglutination (HA) and HI tests. Antigen titers varied by eightfold or more in different diluent systems; titers were generally higher at pH 6.2 than at pH 7.2. A diluent without Ca(2+) gave antigen titers as high as those obtained in diluents with added Ca(2+) ions. Antibody titers also varied in different diluent systems. HEPES diluents at pH 6.2 gave higher antibody titers than those obtained in other diluents, but occasional "false-positive" inhibition reactions were seen. Kaolin suspended in borate saline at pH 9.0 effectively removed inhibitor from sera without absorbing specific antibody, but at pH 7.3 it removed various amounts of specific antibody. Antibody titers of sera treated with kaolin at pH 9.0 were similar to those of sera treated with heparin-MnCl(2); treatment with dextran sulfate-CaCl(2) gave lower antibody titers. Antigens varied widely in sensitivity for detecting HI antibody and in the ability to detect diagnostically significant increases in antibody. Sensitivity in detecting antibody was not related to the HA titer of the antigens. Tween-ether-treated antigens gave lower antibody titers but were more reliable than corresponding untreated antigens for serological diagnosis of infection.

Animals↗

Indirect hemagglutination test for detection of antibodies to Cytomegalovirus.

An indirect hemagglutination test has been adapted for use with cytomegalovirus. The test is highly sensitive and reproducible. Both immunoglobulin M and immunoglobulin G antibodies can be detected by this method. The hemagglutination reaction can be inhibited by small amounts of homologous antigen. This principle permits early identification of virus isolated from diagnostic specimens.

Adult↗

Trypsinized human group O erythrocytes as an alternative hemagglutinating agent for Japanese encephalitis virus.

Trypsinized human group O erythrocytes were found to be a suitable alternative to gander cells in hemagglutination (HA) and hemagglutination inhibition (HAI) tests for Japanese encephalitis (JE) virus. In the HAI test, no cross-reactions against JE virus were observed with immune sera containing antibody to taxonomically related or unrelated viruses, with mouse brain antigen, or with nonantibody serum inhibitors; specific antibody rise could be detected in an immunized rabbit. Gander and trypsinized human group O cells gave comparable titers in the HAI test, but the latter were preferable since (i) they required less challenging HA antigen, being more sensitive to agglutination by JE virus, and (ii) all human and some animal sera investigated were devoid of natural agglutinins for these cells, thereby eliminating or reducing the need for prior adsorption with packed cells.

ABO Blood-Group System↗

Serological comparison of canine rotavirus with various simian and human rotaviruses by plaque reduction neutralization and hemagglutination inhibition tests.

By the plaque reduction neutralization test, the CU-1 strain of canine rotavirus was similar, if not identical, to three strains (no. 14, no. 15, and P) of the tentatively designated third human rotavirus serotype. In addition, strain CU-1 demonstrated a one-way antigenic relationship with two other strains (M and B) of the third human rotavirus serotype. The CU-1 strain of canine rotavirus hemagglutinated human group O, rhesus monkey, dog, sheep, and guinea pig erythrocytes. A two-way antigenic relationship between canine (CU-1) and simian (MMU 18006 and SA11) rotaviruses demonstrated previously by the plaque reduction neutralization test was confirmed further with two additional isolates (A79-10 and LSU 79C-36) of canine rotavirus by the plaque reduction neutralization test and the hemagglutination inhibition test. The CU-1 strain of canine rotavirus, which is known to be distinct from two well-characterized human rotavirus serotypes (Wa and DS-1), was also found to be distinct from the St. Thomas no. 4 strain, which is a newly defined fourth human rotavirus serotype. Thus, this canine strain, which is related antigenically to one of four human rotavirus serotypes, is another example of an animal rotavirus which shares serotype specificity with a human rotavirus.

Animals↗

Effect of urea on the hemagglutinating and complement-fixing antigens of type 2 Dengue virus.

Sephadex G-200 filtration was used for fractionating dengue type 2 (DEN-2)-infected suckling mouse brain (SMB) supernatant fluids. The high-molecular-weight fraction, which was eluted in the void volume, showed no increase in type specificity when tested against immune ascitic fluid to the four prototype dengue viruses. A void-volume fraction obtained after the infected SMB supernatant fluids were treated with urea displayed significant increases in complement fixation (CF) type specificity. Immune ascitic fluid prepared against the more typespecific DEN-2 antigen demonstrated neutralizing ability and greater CF type specificity. When DEN-2 sucrose-acetone-extracted hemagglutinating (HA) antigens were treated with 6 m urea at 37 C for various time intervals, all HA antigen was destroyed in 15 min. Urea treatment of infected SMB supernatant fluids indicated that the CF antigens were more stable to the effect of urea than were the HA antigens. After urea treatment of the SMB supernatant fluids, the CF type specificity increased as the hemagglutination inhibition titer decreased.

Animals↗

Preparation and use of erythrocyte-globulin conjugates to Lassa virus in reversed passive hemagglutination and inhibition.

Conditions were defined for functional covalent coupling of anti-Lassa virus globulins to glutaraldehyde-fixed chicken erythrocytes. Tolylene-2,4-diisocyanate in a reaction mixture containing not more than 0.01 M NaCl produced uniformly good conjugates which were used in reversed passive hemagglutination (RPH) and reversed passive hemagglutination inhibition (RPHI) tests to detect Lassa virus antigens in infected cell cultures and specific antigens in Vero cell cultures. Identical results were obtained with this method and with immunofluorescent-antibody (IFA) staining in the detection and identification of Lassa virus isolated from human and rodent specimens from West Africa. The RPHI method was equal to IFA for serological diagnosis of acute human Lassa virus infection and superior to IFA, complement fixation, and a radioimmunoassay procedure for detection of Lassa virus antibodies in a human population where this infection is endemic.

Animals↗

Reactivity of nonsensitized control erythrocytes in rubella passive hemagglutination assays.

Two commercially available, nonsensitized control erythrocytes for use in rubella antibody passive hemagglutination assays detected nonspecific reactions in 10 of 600 (1.7%) and 9 of 500 (1.8%) sera tested. Reactive sera were positive with one or the other cell type, but not both. The probability of obtaining a false-positive result owing to a nonspecific passive hemagglutination reaction was estimated to be 0.17% for either cell type.

Erythrocytes↗

Hemagglutination patterns of Aeromonas spp. in relation to biotype and source.

Aeromonas spp. show patterns of hemagglutination with human group O cells in the presence of fucose, galactose, and mannose. These patterns are related to biotype as well as to the source of isolates. There was good correlation between hemagglutination pattern and the presence of diarrhea among strains isolated in Western Australia, which was the only source with adequate data for classification of children with an without diarrhea. Most of the environmental and other nonfecal isolates produced patterns different from those in strains associated with diarrhea. These results suggest that hemagglutinins should be considered with enterotoxins as virulence factors in Aeromonas spp.

Aeromonas↗

Passive hemagglutination test for measles immunity and serodiagnosis.

A passive hemagglutination (PHA) test for measles was evaluated in comparison with hemagglutination inhibition (HI) and neutralization (NT) tests. The PHA test determines exclusively the level of antibody directed to the hemagglutinin protein of measles virus. The ratio of PHA to HI titer was 1 to 32 (geometric mean, 6.5) for the first 5 weeks of infection but declined to near unity thereafter. It gradually increased again to 4 to 32 (geometric mean, 11.7) over several years. The initial high PHA titer relative to the HI titer was most likely due to the presence of the immunoglobulin M antibody known to be efficient in agglutination, because 2-mercaptoethanol (2ME) treatment of sera reduced the PHA titer to a level similar to that of the HI titer. The PHA titer in sera obtained after the convalescent phase was insensitive to 2ME, and the relative increase in the PHA over the HI titer was presumably a result of increased antibody avidity. In some individuals, the HI titer fell to below detectable levels several years after either natural infection or vaccination, but the PHA as well as the NT titer remained positive. The PHA titer was therefore a more reliable and more sensitive indicator of immune status against measles than the HI titer. The decrease in PHA titer by 2ME treatment provided evidence of a current or very recent infection. PHA was found to be useful both for assessing immunity status and for serodiagnosis.

Adolescent↗

Analysis of the hemagglutination activity domains of the Venezuelan equine encephalomyelitis and eastern equine encephalomyelitis viruses.

The hemagglutination (HA) domains of the Venezuelan equine encephalomyelitis (VEE) and the eastern equine encephalomyelitis (EEE) viruses providing the interaction of virions and red blood cells were studied with the use of a panel of 17 hemagglutination inhibition (HI) monoclonal antibodies (MAbs). A highly conserved domain (C domain) forming alphavirus-group-reactive MAbs was identified in the E2 protein of the VEE and EEE viruses. These MAbs inhibited HA of the western equine encephalomyelitis, Semliki Forest, Sindbis, Getah, Aura, Chikungunya and Pixuna viruses. The involvement of amino acid residues 59 and 232 in the formation of the C region was demonstrated by sequencing the gene encoding the E2 protein of three escape variants of the VEE virus.

Alphavirus↗

Reversed passive hemagglutination and inhibition with Rift Valley fever and Crimean-Congo hemorrhagic fever viruses.

Reversed passive hemagglutination (RPHA) tests were performed by coating glutaraldehyde-fixed and tannic acid-treated sheep erthrocytes with antibodies to Rift Valley fever (RVF) and Crimean-Congo hemorrhagic fever (CCHF) viruses. Cells coated with crude antibody, in the form of mouse immune ascitic fluid, reacted with high specificity and sensitivity in RPHA tests with live or inactivated virus antigens. In inhibition (RPHI) tests, RVF antibody titers in human and sheep sera were similar to those determined by hemagglutination inhibition test. RVF virus antigen could be detected in viremic sheep sera by the RPHA technique. RPHI antibody titers to CCHF virus in human and hare sera were similar to indirect immunofluorescence (IF) titers, but sheep and cattle sera with RPHI titers of 1:32 or less were negative in IF tests.

Animals↗

Evidence of exposure of waterfowl and other aquatic birds to the hemagglutinating duck adenovirus identical to EDS-76 virus.

Serum and fecal samples from 12 species of aquatic birds were studied for evidence of exposure to a hemagglutinating duck adenovirus (DAV). DAV is serologically indistinguishable from egg-drop syndrome-76 virus. A total of 285 serum samples were tested by the hemagglutination-inhibition (HI) test. Forty-two percent of the birds had HI antibodies, with titers ranging from 8 to 256. Wild ducks showed the highest frequency of antibodies (56%) while in coots and grebes, antibody was less frequent, 33% and 26%, respectively. Attempted virus isolations from 79 fecal samples were unsuccessful. The data support the hypothesis that DAV is indigenous in wild duck populations and suggest that infection and viremia are limited in time and occur at a very early age.

Adenoviridae↗