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PubMed1973

Evaluation of passive hemagglutination, solid-phase radioimmunoassay, and immunoelectroosmophoresis for the detection of hepatitis B antigen.

Sensitivity and specificity of passive hemagglutination (RCA), solid phase radioimmunoassay (RIA), and immunoelectroosmophoresis (IEOP) were compared under experimental and clinical conditions. In dilution experiments with sera containing hepatitis B antigen (HB Ag) of known subtypes, the sensitivity for an ad subtype serum was RIA (1), RCA (1/2), IEOP (1/256) and for an ay subtype serum RCA (1), RIA (1/8), IEOP (1/128). An evaluation of the National Institutes of Health, Division of Biologics Standards test panel number 2 demonstrated HB Ag in 34 of 60 samples by RIA, in 33 by RCA, and in 25 by IEOP. HB Ag was detected in 57.5% of 200 outpatients with a tentative diagnosis of hepatitis by RIA, in 54% by RCA, and in 42.5% by IEOP. In 1,661 volunteer blood donors, 13 (0.78%) were "positive" for HB Ag by RIA, 11 (0.66%) by RCA, and 3 (0.18%) by IEOP. However, absorption experiments indicated that at least six of the above RIA positive and five of the RCA positive sera exhibited nonspecific positive reactions.

Absorption↗
PubMed1973

In vitro production of Clostridium perfringens enterotoxin and its detection by reversed passive hemagglutination.

The reversed passive hemagglutination (RPHA) test yielded a positive reaction in 2 h with as little as 0.5 ng of purified Clostridium perfringens enterotoxin (CPE) per ml as well as with cultures of some C. perfringens grown in Duncan-Strong (DS) medium. This method is the most sensitive, the simplest, and the fastest among all reported. The time course of CPE production of Clostridium perfringens NCTC 8798 in DS was investigated by RPHA. CPE in culture was detectable at 4 h, increased gradually, reached a maximum at 12 to 14 h, and remained at a high level of 20 mug/ml through 48 h of incubation. CPE synthesized within cells is released easily by sonic disruption of young cultures and by aging the cultures 20 h or more. Heat shock of the cell inoculum was essential for CPE production by C. perfringens in DS.

Bacteriological Techniques↗
PubMed1973

Evaluation of a hemagglutination test for human leptospirosis.

An indirect hemagglutination test for the diagnosis of leptospirosis is described; the test uses a soluble antigen from serotype patoc to sensitize sheep erythrocytes which are then fixed with glutaraldehyde. Evaluation of this procedure indicates that it is more reliable than the conventional macroscopic agglutination test and, in contrast with both microscopic and macroscopic agglutination tests, is positive only with sera from persons with current leptospiral illness. The test is simple and convenient and sensitized fixed cells may be stored for at least a year. In comparison with the macroscopic and microscopic tests, only a single antigen is required.

Animals↗
PubMed1974

Use of stable, sensitized cells in indirect micro hemagglutination test for amebiasis.

Human "O" cells were fixed with pyruvic aldehyde, treated with tannic acid, and fixed with glutaraldehyde. The cells were sensitized with amoeba antigen and stored in a refrigerator. The sensitized cells were used periodically for the indirect hemagglutination test with a battery of sera from patients with intestinal amebiasis and confirmed and unconfirmed amebic liver abscess, and also from negative controls. The same battery was tested with cells sensitized with different batches of antigen and also with fresh sheep cells. None of the cells showed any reaction with negative control sera. The fixed cells remained sensitive and stable throughout the study. Reproducibility of the titers with the fixed cells within each day and from day to day was satisfactory. The titers with fixed human "O" cells were slightly lower than were the titers with fresh sheep cells. The advantages of using stable, sensitized cells are pointed out.

Aldehydes↗
PubMed1974

Stable mycoplasma antigen preparations for indirect hemagglutination tests.

In immunological studies of mycoplasmas, the use of glutaraldehyde for the fixative makes it possible to use erythrocytes from commercially available defibrinated sheep blood. It eliminates the necessity of having to screen blood from individual sheep to obtain a suitable source of erythrocytes, as when employing tannic acid for fixation and sensitization. The chemical bonding of soluble mycoplasma proteins to glutaraladehyde-fixed sheep erythrocytes by bis-diazotized 3,3'dimethoxy derivative, benzidine, yields preparations that are satisfactory antigens for performing the indirect hemagglutination test by the microtiter technique. The antigenic preparations are satisfactory for use after storage at 4 or -10 C for many months. Incorporation of 5% glycerine in the final suspending milieu makes it possible to obtain uniform suspensions of the fixed and sensitized sheep erythrocytes after freezing and after repeated freezing and thawing. Proteins from Mycoplasma arthritidis and M. hominis have been coupled to glutaraldehyde-fixed erythrocytes by diazotization. The last mentioned preparation detected the presence of antibodies in titers greater than 1:10 in 37% of 237 pregnant women whose ages ranged between 20 and 30 years. There was no correlation between the presence of specific antibodies in the blood and the isolation of M. hominis from the cervical canal.

Adult↗
PubMed1974

Compatibility of trypsin-modified human erythrocytes in the rubella hemagglutination-inhibition test employing three serum treatment procedures.

Results of comparative tests using trypsin-modified human type O erythrocytes and cells from newly hatched chickens with three standard serum treatment methods for rubella hemagglutination-inhibition techniques are reported. The kaolin, heparin-manganous chloride and dextran sulfate-calcium chloride methods could all be used with both cell types. Reproducibility with heparin-manganous chloride and dextran sulfate-calcium chloride was excellent with both cell types. Both methods gave generally higher antibody titers than the kaolin procedure. However, the use of human cells resulted in a more sensitive system than chicken cells with all serum treatment methods.

Animals↗
PubMed1974

Indirect hemagglutinating antibody response to Herpesvirus hominis types 1 and 2 in immunized laboratory animals and in natural infections of man.

Indirect hemagglutinating (IHA) antibody responses to Herpesvirus hominis types 1 and 2 (HVH-1 and HVH-2) were compared to complement-fixing and neutralizing antibody responses in immunized laboratory animals (rabbits, guinea pigs, and hamsters) and in natural infections of man. With the immunized animals, type specificity was seen only in the IHA test and only with antisera produced in hamsters and in the rabbits immunized with HVH-2. In human nongenital infections (considered to be caused predominately by HVH-1), IHA and neutralizing antibodies developed at about the same rate and reached approximately the same levels for HVH-1 and HVH-2. IHA titers tended to be higher than neutralizing antibody titers for both virus types. In genital infections (considered to be caused predominately by HVH-2), there was a rapid IHA antibody response to HVH-2, and the early HVH-2 antibody demonstrable by IHA, but not by neutralization tests, was found to be immunoglobulin M in nature. In genital infections, IHA titers for HVH-2 were markedly higher than neutralization titers, but there was no pronounced difference in neutralizing the IHA antibody titers for HVH-1. Several patients with genital infections fialed to develop IHA antibody for HVH-1. The IHA test possessed no greater sensitivity than did complement fixation or neutralization tests for serodiagnosis of HVH infections. Despite the fact that a number of patients with genital infections produced IHA antibody only for HVH-2, the test was no more effective than the neutralization test in providing a type-specific serodiagnosis of infection, due largely to the fact that the rapid IHA antibody response to HVH-2 prevented demonstration of a further, significant antibody titer increase in a number of cases.

Animals↗
PubMed2003

Comparison of complement fixation and hemagglutination inhibition assays for detecting antibody responses following influenza virus vaccination.

Complement fixation (CF) was compared to hemagglutination inhibition (HI) as a method for identifying antibody responses to influenza virus vaccination. CF assays were performed at two different laboratories using paired (pre- and postvaccination) sera from 38 vaccinated laboratory employees; HI assays were performed at a third laboratory. As expected, most vaccinees (31/38 = 82%) responded to at least one of three influenza virus antigens as measured by HI. In contrast, only 21% (8/38) of vaccinees showed a response by CF at laboratory 1, and only 29% (11/38) showed a response by CF at laboratory 2. These findings indicate that due to low sensitivity, CF assays should not be used to assess the antibody response to influenza virus vaccination.

Antibodies, Viral↗
PubMed2003

Antibody responses determined for Japanese dengue fever patients by neutralization and hemagglutination inhibition assays demonstrate cross-reactivity between dengue and Japanese encephalitis viruses.

Titers of antibodies to infecting dengue virus serotypes determined by serum neutralization assay were higher than those of antibody to Japanese encephalitis (JE) virus in Japanese dengue patients after disease day 8. Titers of antibody to dengue virus antigens determined by hemagglutination inhibition (HI) assay were higher in only 1 of 23 serum specimens after disease day 11. Thus, the neutralization test is more reliable than the HI test for serological diagnosis of dengue in countries where JE vaccination is widely used or JE is endemic.

Adolescent↗
PubMed1998

Comparison of neutralizing and hemagglutination-inhibiting antibody responses to influenza A virus vaccination of human immunodeficiency virus-infected individuals.

A neutralization enzyme immunoassay (N-EIA) was used to determine the neutralizing serum antibody titers to influenza A/Taiwan/1/86 (H1N1) and Beijing/353/89 (H3N2) viruses after vaccination of 51 human immunodeficiency virus (HIV) type 1-infected individuals and 10 healthy noninfected controls against influenza virus infection. Overall, the N-EIA titers correlated well with the hemagglutination-inhibition (HAI) titers that were observed in the same samples in a previous study (F. P. Kroon, J. T. van Dissel, J. C. de Jong, and R. van Furth, AIDS 8:469-476,1994). The N-EIA appeared to be more sensitive than the HAI test. Significantly more fourfold or higher rises in N-EIA titer and higher mean N-EIA titers occurred in HIV-infected individuals with > or =200 CD4+ cells per microl than in those with <200 CD4+ cells per microl.

Adult↗
PubMed2006

Neutralizing and hemagglutination-inhibiting activities of antibodies elicited by the 2004-2005 influenza vaccine against drifted viruses.

Evaluation of the antibody responses induced by the 2004-2005 influenza vaccine strain against the homologous variant, the 2004-2005 field isolates, and a previous circulating strain showed that a correlation between neutralizing and hemagglutination-inhibiting activities exists only when the antigen is very close to the vaccine strain.

Antibodies, Viral↗
PubMed1974

Reactivity of envelope, capsid, and soluble antigens of Herpesvirus hominis types 1 and 2 in the indirect hemagglutination test.

Envelope, capsid, and soluble antigens of Herpesvirus hominis (HVH) types 1 and 2 were compared to crude antigens (disrupted HVH-infected cells) for potency, type-specificity, and diagnostic value in the indirect hemagglutination (IHA) test. The envelope appeared to be the predominant component reacting in the IHA test, but recurrent HVH infections increased the reactivity of human sera with capsid antigens. The soluble antigens reacted in the IHA test with HVH immune animal sera, but very few convalescent-phase human sera showed reactivity with soluble antigens. Overall, none of the subunit antigens showed greater type-specificity than did crude antigens in IHA tests with immune animal or convalescent-phase human sera. Recurrent infections with type 1 or type 2 viruses tended to broaden heterotypic reactivity of the patients' sera with both crude and subunit antigens, even in patients showing only a single type of antibody by IHA inhibition. Subunit antigens were no more sensitive than crude antigens in demonstrating significant IHA antibody titer rises for serodiagnosis of herpesvirus infections, and they generally had to be used at lower working dilutions than crude antigens.

Animals↗
PubMed1975

Identification of measles virus-specific hemolysis-inihibiting antibodies separate from hemagglutination-inhibiting antibodies.

The occurrence of antibodies giving hemolysis inhibition (HLI) but not hemagglutination inhibition (HI) was examined in human convalescent and rabbit hyperimmune sera. HI antibodies, which through their interaction with hemagglutinin components display HLI activity, were removed by absorption with Tween 80-ether (TE)-treated measles virus material. This absorption did not change the titer of non-HI HLI antibodies. After removal of HI antibodies from 16 late measles convalescent sera and three batches of gamma globulin. HLI antibody titers showed a two- to eightfold reduction. The titers of neutralizing antibodies were reduced from 1/4 to 1/20 of the original titers. There was a good correlation between the titers of neutralizing and HLI antibodies both in sera from which HI antibodies had been removed by absorption and in sera spontaneously showing markedly higher HLI than HI antibody titers. HLI antibodies with these characteristics could be identified in HI tests when whole virus instead of TE-treated material was used an antigen and anti-antiserum was added to the tests. In contrast to the situation in human sera, antibodies remaining after removal of HI antibodies from rabbit hyperimmune sera against purified virus particles were detectable in neutralization and HLI tests only in the presence of anti-antiserum. However, virus particles from which the major fraction of all envelope projections had been removed by treatment with 0.004% trypsin induced the production of non-HI HLI antibodies active also in the absence of anti-antiserum. TE and formalin treatment destroyed the hemolytic activity of virus preparations and also their capacity to induce a production of non-HI HLI antibodies.

Absorption↗
PubMed1976

Purification of hepatitis A antigen from feces and detection of antigen and antibody by immune adherence hemagglutination.

Hepatitis A antigen (HA Ag) was purified from feces collected during acute illness from patients with naturally occurring viral hepatitis, type A. Positive fecal specimens were identified by immune electron microscopy, but for detection of HA Agduring purification immune adherence hemagglutination (IAHA) and microtiter solid-phase radioimmunoassay were used. Isopycnic banding in cesium chloride, rate-zonal separation in sucrose, and preparative zonal electrophoresis were used in various combinations for successive purification, and the purified antigen was successfully used in a test for antibody by IAHA. Seronconversions to HA Ag were demonstrated by IAHA in 20 instances of hepatitis A virus infection, but in none of six cases of type B hepatitis or three cases of post-transfusion hepatitis unrelated to heaptitis A or B viruses, nor in two individuals without hepatitis. In addition, the temporal pattern of antibody development during type A hepatitis was studied in serial sera from an experimentally infected chimpanzee. Antibody titers by IAHA correlated well with antibody ratings determined by immune electron microscopy.

Antibodies, Viral↗