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PubMed1984

[Standardization of a microtechnic for passive hemagglutination to detect antibodies against infectious bovine rhinotracheitis].

A micromethod for passive Hemagglutination is described for the detection of IBR antibodies in serum samples of nine herds. The samples correspond to different zones of the province Buenos Aires. In animals without clinical signs the proportion of positives was 43.60%; in herds with signs consistent with the disease, this value attained 66.4% positivity (Tables 1 and 2). The feasibility of this method is discussed. Some variants introduced into the original technique are described.

Animals↗
PubMed1980

[Use of the aggregate hemagglutination method for determining cholera vibrio neuraminidase].

The suitability of the aggregate hemagglutination test for neuraminidase determination in biological fluids has been evaluated. This test allows one to determine neuraminidase at a concentration of 2--4 mg/ml. The sensitivity of this test is more than 2 orders higher than that of gel diffusion technique and the thiobarbituric assay. The test has revealed that neuraminidase produced by Vibrio cholerae is not immunologically related to other kinds of neuraminidase produced by Corynebacterium diphtheriae, Clostridium perfringes and Erysepelothrix insidiosa.

Antigens, Bacterial↗
PubMed1981

Use of the immune adherence hemagglutination test for titration of breast cancer patients' sea cross-reacting with purified mouse mammary tumor virus.

Ninety-two sera from patients with breast cancer, 42 sera from patients with neoplastic diseases other than breast cancer and 59 sera from apparently healthy women were examined by means of the immune adherence hemagglutination (IAHA) test using purified mouse mammary tumor virus (MMTV) fron RII mouse milk. It was found that 36.4% (34/96) of the sera from breast cancer patients, 7.1% (3/42) of the sera from patients with other neoplastic diseases and 5.1% (3/59) of the sera from apparently healthy women showed a positive reaction. Among the IAHA positive sera from breast cancer patients, 82.9% (29/35) showed a titer of more than 1:16. On the other hand, none of the positive sera from patients with cancers other than breast cancer showed a titer of more than 1:16. The sera from 4 breast cancer patients, which showed a positive reaction with RII MMTV in the IAHA test, were tested to examine the specificity of the reaction by using milk samples from sources other than RII mice, including C57BL mice, dogs, cattle and humans. None of the 4 sera showed a positive reaction with milk samples from sources other than the RIII mouse.

Adult↗
PubMedDate not supplied

Evaluation of the hemagglutination-inhibition test for measuring the response of chickens to avian infectious bronchitis virus vaccination.

An infectious bronchitis virus (IBV) hemagglutination-inhibition (HI) test was used to assay serum-antibody titers after IBV vaccination of IBV-susceptible specific-pathogen-free broilers and commercial layers. Three-week-old broilers were vaccinated via eye-drop with IBV strains that represent the antigenic spectrum of commercial vaccines--Holland, Massachusetts 41 (41 Ms), Connecticut 46, Florida 18288, or JMK strain--and revaccinated 3 weeks later with either the same or a heterologous strain. Weekly serum samples were tested by IBV HI with homologous and heterologous antigens. Vaccinates, except for those vaccinated with the Holland strain, were HI-positive with homologous but not heterologous antigens by 1 to 2 weeks postvaccination. Sixteen-week-old IBV-vaccinated commercial layers were revaccinated with IBV Holland 52 (H 52) strain and subsequently infected with Arkansas 99 (Ark 99) and SE 17 strains. In contrast to the limited HI cross-reactivity of serum from IBV-vaccinated broilers, there were extensive cross-reactions in HI tests with 41 Ms, H 52, Ark 99, and SE 17 antigens of revaccinated layers. These results demonstrate that the IBV HI test is more strain-specific than previous reports indicate, especially when the test samples are from early postvaccination.

Animals↗
PubMed1983

Graded hemagglutination inhibition for quantification of human fetal hemoglobin.

We describe a hemagglutination system for quantification of fetal hemoglobin (HbF), especially at low concentrations. This system consists of three components: (a) sheep erythrocytes coated with purified HbF, (b) sheep antiserum of human HbF made specific for the gamma-chain, and (c) human blood lysate with a known concentration of HbF as the standard. We can measure the whole range of HbF proportions in human blood, from 100% down to 0.01%. Statistical analysis showed that our measurements agreed with those made by the methods of alkali denaturation and radial immunodiffusion. Reproducibility of the method is shown by the low CVs for six repeated measurements. Low concentrations of HbF not measurable by other methods are easily assayed. We offer examples of the clinical utility of data on low concentrations of HbF.

Aged↗
PubMed1982

Indirect hemagglutination for the diagnosis of Heterobilharzia americana infections in dogs.

An indirect hemagglutination test was developed for the diagnosis of canine schistosomiasis. Specific-pathogen-free dogs and dogs infected with a variety of other helminths were experimentally infected with Heterobilharzia americana. Sera collected at various times after the dogs were exposed to cercariae were evaluated. Glutaraldehyde-fixed sheep erythrocytes sensitized with whole adult worm antigens provided a stable antigen substrate when tested over a period of months. The test produced sensitive and specific diagnosis at an appropriate serum dilution. Serum dilutions less than 1:80 from dogs with other helminth infections resulted in nonspecific agglutination. By postexposure day 60, coinciding with the end of the prepatent period, serum titers of dogs infected only with H americana equaled, and then surpassed, those of the other dogs.

Animals↗
PubMed1982

[Analysis of the serodiagnostic reactions of the bacterium Clostridium butyricum strain CNRZ 528 using passive hemagglutination].

For serologic tumour diagnostics a specific proof of antibodies against rods of Clostridium butyricum CNRZ 528 by passive hemagglutination has been described. We were able to prepare an antigen from the cell extract (crude-antigen) of rods without flagella by gel filtration, that after fixation on the surface of human-erythrocytes, stabilized by tannin or glutaraldehyde, agglutinates specifically with rod-antibodies of rabbit-hyperimmunserum. Simultaneously however, there was no cross-reactivity with spore-antibodies of rabbit-hyperimmunserum. Sensitivity and specificity of this test are dependent on an optimal dose of antigen bound to the surface of the erythrocytes. The optimization of the assay has to be achieved by the testing of patients sera.

Animals↗
PubMedDate not supplied

Investigations on the presence of antibodies to papova viruses in patients with different forms of cancer and in other categories of patients or apparently healthy subjects. Note II. Hemagglutination-inhibiting serum antibodies to BK virus.

Hemagglutination-inhibiting serum antibodies to BK virus (BKV were detected in patients with different forms of cancer and in blood donors (positivity percentages: 67.04 and 57.78, respectively). No such antibodies were found in a group of children 1 to 14 years of age. The data point to the presence of a latent human infection with BKV in the population of Romania.

Adolescent↗
PubMedDate not supplied

Investigations on the presence of antibodies to papova viruses in patients with different forms of cancer and in other categories of patients or apparently healthy subjects. Note III. Hemagglutination-inhibiting serum antibodies to polyoma virus.

Hemagglutination-inhibiting antibodies to polyoma virus could be detected in sera from apparently healthy subjects, patients with nonmalignant respiratory diseases and patients with different forms of cancer. The positivity percentages (12.97, 12.92 and 16.36, respectively) and the titers recorded were lower than in the case of BK virus. The results obtained suggest a slight antigenic relationship between polyoma and BK virus.

Adolescent↗
PubMed1982

Indirect hemagglutination antibody titers for Entamoeba Histolytica in dried filter paper blood and sera.

Serum and elutates from dried filter paper bloods from 404 patients with and without amebic liver abscess (ALA) were tested for Entamoeba histolytica antibodies by the indirect hemagglutination test. Eighty-nine percent of the sera from patients with ALA were positive (titers greater than or equal to 1 : 128) and 94% of those without ALA negative. Fifty-five percent of the filter paper bloods from ALA patients were positive. Overall agreement was 80% for positives and negatives and the correlation coefficient between the methods showed a linear relationship (r = 0.77). Although the quantity of sera available from dried filter paper bloods is limited and some antibody activity may be lost, the method should be of value in seroepidemiologic studies where venous bloods are not easily obtained.

Adolescent↗
PubMedDate not supplied

Preparing hemagglutinating antigen from isolates of infectious bronchitis virus.

The hemagglutinating (HA) activity of 14 strains of infectious bronchitis virus (IBV) was investigated. The optimal conditions for IBV antigen preparation include inoculation of 10- or 11-day-old specific pathogen-free embryonated eggs and incubation for 30 hours at 37 C. Embryos were inoculated via the allantoic cavity with 0.1 ml of a low embryonic passage of the virus (10(7) to 10(8) EID50/ml). Allantoic fluid was harvested and pooled, and a 100-fold concentration of virus particles was achieved by centrifugation for 3 hours at 30,000 x g. Virus pellets were resuspended in Tris-hydrochloride buffer containing 3 units of phospholipase-C (type-1) enzyme/ml and incubated for 2 hours at 37 C. All IBV strains tested demonstrated positive HA activity with chicken red blood cells. The antigen was stored in liquid state or lyophilized at 4 C.

Animals↗
PubMed1983

Detection of antibody to BK virus by enzyme-linked immunosorbent assay compared to hemagglutination inhibition and immunofluorescent antibody staining.

The enzyme-linked immunosorbent assay (ELISA), hemagglutination inhibition test (HAI), and immunofluorescent antibody staining (IFAS) test were compared for their ability to detect IgG antibody to BK virus (BKV). Antibody titers to BKV antigen were determined for 35 sera by each of the three assays. The ELISA tests agreed with either HAI or IFAS with regard to BKV antibody status in all but one serum. On this basis the HAI accounted for four discordant determinations while IFAS was not in agreement in three cases. Geometric mean titers of ELISA and HAI were similar but greater than that of IFAS. The ELISA is particularly well suited for use in large-scale seroepidemiological studies since automated spectrophometric equipment can provide rapid, objective, numerical data.

BK Virus↗
PubMedDate not supplied

Comparison of the hemagglutination-inhibition test with the constant-virus diluting-serum microneutralization test for detecting antibody to avian infectious bronchitis virus.

An evaluation was made of the hemagglutination-inhibition test and the constant-virus diluting-serum microneutralization test, two serological techniques that are used in detecting antibodies to avian infectious bronchitis. The repeatabilities and sensitivities of both tests were similar. Both tests compared favorably when testing high-titered antisera, but antibody titers detected varied when low-titered sera were used.

Animals↗
PubMedDate not supplied

Observations on the preparation and stability of infectious bronchitis virus hemagglutination antigen from virus propagated in chicken embryos and chicken kidney cell cultures.

A total of 166 infectious bronchitis virus (IBV) hemagglutination (HA) antigen preparations were made during a 30-month period from allanto-amnionic fluid (AAF) from chicken embryos inoculated with 10 different IBV strains (Mass 41, Conn 46, H52, Florida 18288, Ark 99, JMK, T, Holte, EF, SE17). Antigens were prepared by inoculating 9- or 10-day-old embryos with 10(5.0) to 10(6.5) EID50 IBV, harvesting AAF after a 30-hour-postinoculation incubation, and phospholipase C (PLC) treatment of virus concentrated by pelleting from the AAF. Longer (48 hr) incubation times were tried, but production of H52 HA antigen was successful only from AAF harvested after 30 hours of incubation. AAF from JMK-infected embryos had lower infectivity titers and frequently yielded lower HA antigen titers than the other strains. The treatment of AAF with fluorocarbon did not enhance or diminish HA activity but did yield clearer antigens by removing extraneous material. Polyethylene glycol precipitation of virus was an acceptable alternative to pelleting virus at 39,000 X g. Inactivation of IBV with 0.1% betapropiolactone did not affect HA activity, whereas inactivation with 0.1% formalin caused a marked reduction in HA titer. Different buffer formulations including phosphate, tris, or HEPES were tried to optimize the conditions for PLC treatment of virus concentrate, but there were no apparent differences in the antigens prepared in the different buffers. The HA antigen preparations were stored and were stable at 4 C. Antigen titers of greater than or equal to 64 after storage for 20 months or longer were not uncommon. Addition of merthiolate as a preservative had no deleterious effect on HA activity. Antigen stability appeared to be enhanced by incorporating EDTA in buffer for virus pellet recovery and during enzyme treatment. Attempts to produce HA antigens from cell-culture-adapted virus propagated in chicken kidney cells were less satisfactory. An acceptable HA antigen was prepared from only two (Mass 41, SE17) of the seven strains that were tried. Virus propagation in chicken embryos is the better method of the two for IBV HA antigen production. Aside from the need to concentrate virus and treat the concentrate with PLC, there appeared to be considerable latitude in the procedures that can be used to make acceptable IBV HA antigens.

Allantois↗
PubMed1983

Comparison of hemagglutination and competitive enzyme-linked immunosorbent assay procedures for detecting canine parvovirus in feces.

A competitive enzyme-linked immunosorbent assay (ELISA) for detection of canine parvovirus (CPV) antigen in fecal samples was developed. Fecal samples were tested by ELISA and a direct hemagglutination assay, and the results compared. The tests gave the same results in 83% and 88% of the fecal samples, depending on whether the samples were treated with chloroform. The discrepancies were due to the fact that each test detected virus in certain fecal samples that was not detected by the other. The use of a monoclonal anti-CPV conjugate resulted in a highly specific test. The performance of the ELISA as a competitive assay also increased the reproducibility and sensitivity over that which could be obtained from a classic sandwich-type procedure.

Animals↗
PubMed1984

[Value of indirect immunofluorescence and passive hemagglutination reactions in the detection of urogenital trichomoniasis].

The indirect fluorescent antibody (IFAT) and indirect hemagglutination tests (IHA) were performed in 375 patients attending the venereal disease clinic (CHU of ROUEN). In addition, wet mount, acridine orange stain and culture were used to detect Trichomonas vaginalis in genital discharge. Significant levels of antitrichomonas antibodies were present in sera, in 44 per cent of women with vaginal discharge and 23 p. cent of men with urethritis or prostatitis, whereas no evidence of trichomonal infection was demonstrated. These data suggest that IFAT and IHA tests may be useful to improve the diagnosis of uro-genital trichomoniasis.

Adult↗
PubMed1983

[Comparative evaluation of different meningococcal group-specific erythrocyte diagnostic kits in passive hemagglutination test].

The comparative evaluation of the sensitivity and specificity of serogroup A, C, Y meningococcal antigenic preparations obtained by different methods was carried out by means of the passive hemagglutination test. In case of group 0 (I) human red blood cells sensitized with serogroup A and C vaccinal preparations obtained by Gotschlich's method (designated as A-1 and C-1) were used. In the other case formalin-treated sheep red blood cells sensitized with group-specific polysaccharides obtained by alcohol precipitation from the cultural fluid of group A, C and Y meningococci with subsequent heating (designated as A-2, C-2, Y) were used. Titrations with commercial immune rabbit sera showed that both variants of the antigenic preparations were similar in their specificity and sensitivity. In patients with the symptoms of meningitis the diagnostic titer was 1:40 for preparations A-1, C-1 and 1:80 for preparations A-2, C-2 and Y. The results of the examination of 164 patients (220 serum specimens) demonstrated that these preparations were of equal diagnostic value.

Antibodies, Bacterial↗
PubMedDate not supplied

[Rapid diagnosis of Crimean hemorrhagic fever in the indirect reaction to hemagglutination].

The possibility of using indirect hemagglutination test (IHA) with erythrocyte immunoglobulin diagnosticum for detection of CHF virus in the blood of patients early after the onset was studied. It was shown that viremia could be detected by the IHA test within 2-3 hours provided the blood was collected not later that 8 days after the onset of the disease. The sensitivity of the test for the detection of viremia is as high as that of bioassays. The IHA test may also be used for the detection of CHF virus specific antigen in cadaver materials (blood, organ suspensions) even in those cases when virus isolation is prevented by toxicity of the specimens.

Animals↗