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PubMed1984

Comparison of an enzyme-linked immunoassay with an indirect hemagglutination assay for the detection of antibodies to cytomegalovirus.

Because of the morbidity associated with cytomegalovirus (CMV) infections in neonates, immunosuppressed and transplant patients, there is a need for a rapid, sensitive, and reliable assay for the presence of antibody to CMV. Such an assay would permit the identification of CMV antibody negative blood or plasma for transfusion into these high-risk patients. The indirect hemagglutination assay (IHA) is often employed to detect antibodies to CMV, but results must be interpreted subjectively. Thus, an indirect enzyme immunoassay (EIA) was developed to detect antibodies to CMV, and was compared to IHA. 531 sera from a population of healthy blood donors were assayed by both methods. Compared with IHA the sensitivity of the indirect EIA was 98.5% (267/271) and the specificity was 93.1% (242/260). Antibodies to Epstein-Barr, herpes-zoster, or herpes simplex viruses did not react in the EIA. The EIA and IHA correlated well in the detection of antibodies to CMV in sera from acute CMV infections. The presence of rheumatoid factor did not interfere with the EIA or IHA results. This indirect EIA is a rapid, simple, and reproducible method to identify seronegative sera or plasma.

Adult↗
PubMed1985

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↗
PubMed1989

Avidin-biotin enhanced reverse passive hemagglutination method for detection of HIV antigens.

A new assay for HIV antigens, the avidin-biotin enhanced reverse passive hemagglutination (AB RPHA) test is described which is suitable for detection of infection in cell culture experiments. This assay is simple to perform and economical, and has sensitivity equal to or greater than that of commercial ELISA assays. The coated red cells used for this assay may be stored in the frozen or lyophilized state.

Avidin↗
PubMed1990

Influenza-specific ELISA IgA and IgG predict severity of influenza disease in subjects prescreened with hemagglutination inhibition.

Four influenza A challenge studies were performed over a period of three years using the same dose of one virus pool. The first three studies were conducted two influenza seasons apart from the last study. In all four studies only subjects with screening hemagglutination inhibition (HAI) antibody titers less than or equal to 1:8 in sera were accepted as study subjects. The rate of infection after influenza challenge was 96% (25 of 26) in the first three studies, and only 73% (8 of 11) in the last study (P less than 0.04). The rate of influenza illness in the first three studies was 62% (16 of 26), and only 9% (1 of 11) in the last study (all subjects: P = 0.003; infected subjects: P = 0.01). Even though screening HAI titers were comparable between groups, prechallenge serum influenza-specific IgG titers correlated inversely with respiratory symptoms (P = 0.04). Prechallenge nasal wash influenza-specific IgA titers were lower in subjects who developed influenza illness (P = 0.03). Prechallenge influenza-specific nasal wash ELISA-IgA titers and serum ELISA-IgG titers predicted influenza severity in patients prescreened by HAI during influenza challenge studies.

Adolescent↗
PubMed1990

Hemagglutinating encephalomyelitis virus attaches to N-acetyl-9-O-acetylneuraminic acid-containing receptors on erythrocytes: comparison with bovine coronavirus and influenza C virus.

The receptors for the hemagglutinating encephalomyelitis virus (HEV, a porcine coronavirus) on chicken erythrocytes were analyzed and compared to the receptors for bovine coronavirus (BCV) and influenza C virus. Evidence was obtained that HEV requires the presence of N-acetyl-9-O-acetylneuraminic acid (Neu5,9Ac2) on the cell surface for agglutination of erythrocytes as has been previously shown for BCV and influenza C virus: (i) Incubation of red blood cells with sialate 9-O-acetylesterase, the receptor-destroying enzyme of influenza C virus, rendered the erythrocytes resistant against agglutination by each of the three viruses; (ii) Human erythrocytes which are resistant to agglutination by HEV acquire receptors for HEV after resialylation with Neu5,9Ac2. Sialylation of red blood cells with limiting amounts of sialic acid indicated that strain JHB/1/66 of influenza C virus requires less Neu5,9Ac2 for agglutination of erythrocytes than the two coronaviruses, both of which were found to be similar in their reactivity with Neu5,9Ac2-containing receptors.

Acetylesterase↗
PubMed1984

Detection of serum antibodies to sarcoptic mange mite antigens by the passive hemagglutination assay in pigs infested with Sarcoptes scabiei var. suis.

Development and remission of the clinical signs of sarcoptic mange in pigs experimentally infested once and twice with Sarcoptes scabiei var. suis were described. Encrusted lesions in the ears of pigs and a general dermatitis were observed 3 weeks after infestation, but subsided gradually. Specific antibody activity, in titers averaging 1:16 000 in sera from rabbits immunized with mite extracts and 1:512 in the serum from a pig artificially infested with sarcoptic manage and then twice immunized with mite extract, was detected with the passive hemagglutination assay with tanned bovine red blood cells. Titers ranging from 1:40 to 1:160 were detected in sera from experimentally-infested animals. Only 3 of 75 sera from chronically naturally-infested pigs exhibited antibody titers of 1:40 to the mite extract.

Animals↗
PubMed1992

Early immunodiagnosis of bovine fascioliasis using the specific antigen f2 in a passive hemagglutination test.

An improved hemagglutination (HA) test using the purified specific f2 antigen of Fasciola hepatica has been evaluated with regard to its potential use for early diagnosis of infection. On experimental infection of beef calves 6-8 months of age, f2-specific antibodies were detected 2-4 weeks after inoculation and persisted at high levels during the 28 week experimental period. On natural infection of dairy calves, 6-9 months of age, allowed to graze in infected fields from April to June, 48% of the animals were positive in July although coproscopic analyses were negative. In November all the calves were HA positive but only 24% of them excreted F. hepatica eggs. Beef calves 1-3 months of age allowed to graze with their dams in infected fields continued to suckle their dams and were weakly infected according to HA results. This weak infection was not detected by coproscopical analysis. Colostral f2-specific antibodies persisted for approximately 6 months in the serum of dairy calves allowed to suckle their dams immediately after birth.

Animals↗
PubMed1994

Early immunodiagnosis of caprine fasciolosis using the specific f2 antigen in a passive hemagglutination test.

An improved hemagglutination (HA) test using the purified specific f2 antigen of Fasciola hepatica has been evaluated with respect to its potential use in the diagnosis of caprine fasciolosis. Following experimental infection of 1-year-old goats with a single heavy infection of 300 metacercariae, f2-specific antibodies were detected 2-3 weeks after infection and increased steadily to reach a maximum titer 9 weeks after infection, after which the antibody level declined. In animals receiving multiple infections of a lower dose of 50 metacercariae given at weekly intervals for 6 weeks, f2-specific antibodies were detected 3 weeks after infection and increased to reach a plateau 11 weeks after infection which was maintained until the end of the experiment (15 weeks after infection). Depending on animals and groups, eggs appeared in the feces between 7 and 9 weeks after infection. The HA test may provide valuable information about the early detection of caprine fasciolosis, particularly during the prepatent period.

Animals↗
PubMed1983

Hemagglutinating and hydrophobic surface properties of salmonellae producing enterotoxin neutralized by cholera anti-toxin.

Eleven Salmonella strains known to produce enterotoxin under aerobic culture conditions in deferrated (DF) medium at 37 degrees C were shown to produce enterotoxin with and without aeration at 22, 28, 37 and 42 degrees C. Heat-labile enterotoxin was generally produced with growth temperatures up to 37 degrees C irrespective of aeration. Heat-stable enterotoxin was produced up to 42 degrees C, mainly aerobically, as indicated by infant mouse assay (IMA), by six of the eleven strains tested. Nine strains produced heat-stable rapid permeability factor (RPF) in rabbit skin. Cholera anti-toxin neutralized reactivities of Salmonella heat-labile enterotoxin in four different biological assays. Mixed gangliosides also neutralized this activity in the cell-test systems. With guinea-pig erythrocytes, all strains underwent mannose-resistant hemagglutination (MRHA) irrespective of growth temperatures, i.e. 22 and 37 degrees C or medium, i.e., DF, tryptose soy broth (TSB) and colonization factor antigen (CFA) agar. At both growth temperatures, CFA agar-grown cells of each strain caused MRHA of bovine erythrocytes. Excepting three Salmonella typhimurium strains, DF broth-grown cells gave MRHA of bovine, chicken and human group A erythrocytes, CFA agar-grown cells caused MRHA of chicken and human blood, whereas TSB-grown cells caused few MRHA reactions. Salmonellae producing both heat-stable, (ST) and heat-labile, (LT) enterotoxins adsorbed to Phenyl Sepharose whereas salmonellae that produced only LT enterotoxin did not. The presence of MRHA adhesions did not correlate with cell-surface hydrophobicity. However, mannose-resistant hemagglutinins may occur more commonly among salmonellae than has been previously recognized.

Animals↗
PubMed1984

Evaluation of the indirect hemagglutination assay as a practical serodiagnostic test for mycoplasmal pneumonia of swine.

Sera from swine experimentally or naturally infected with Mycoplasma hyopneumoniae (the etiological agent of mycoplasmal pneumonia of swine, MPS) were tested by the indirect hemagglutination assay (IHA), the enzyme-linked immunosorbent assay (ELISA) and the complement fixation (CF) test. The IHA detected antibody at comparable times and levels to the other 2 serological tests following experimentally-induced infection. In the late antibody response (greater than or equal to 86 days post-infection), the ELISA titres were higher than either the IHA or the CF test. The IHA appeared least satisfactory when it was used to test sera from commercial swine herds. When 1000 sera were tested, the IHA was positive for only 30 (22%) of 135 sera which were positive by the ELISA and the CF test. The IHA titres were low; 20 of the 30 sera had a titre of only 10. The end-points for the IHA were difficult to read for sera of this low titre. The relationship between positive IHA results for the herd sera obtained at necropsy, and the occurrence of gross or microscopic lesions typical of MPS was poor (41 and 50% agreement, respectively). An agreement of 39% was noted between positive IHA results and the localization of mycoplasmal antigens by an indirect immunofluorescence (IIF) test. However, IHA results correlated significantly (P less than 0.05) with gross and microscopic lesions, but not with the IIF test. No significant correlation was noted between the IHA (or the other 2 serologic tests) and the cultural isolation of M. hyopneumoniae or M. flocculare. On the basis of these results, the IHA appears to have limited promise as a practical test for the diagnosis of MPS in commercial swine herds because of the low titres observed, poor correlation of the IHA and other indicators of MPS, the necessarily subjective determination of end-points, and other inherent technical limitations of the test.

Animals↗
PubMed1985

An indirect hemagglutination test for the detection of antibodies to Clostridium chauvoei.

An indirect hemagglutination (IHA) test using sonicated extract as the antigen was developed for the detection of antibodies to Clostridium chauvoei. This antigen can be adsorbed onto glutaraldehyde-fixed sheep red blood cells treated with tannic acid and can be destroyed by trypsin and heat treatment. It corresponded well with the flagella of the organism, when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the gel diffusion test. No serological cross-reactivity was found in the IHA test when the antigen was tested against 4 species of clostridial antibodies. Our results suggest that the IHA test mainly detects antibodies against the flagella of C. chauvoei.

Agglutination Tests↗
PubMed1988

Passive hemagglutination test for detection of antibodies against Taylorella (Haemophilus) equigenitalis in sera of mares.

The passive hemagglutination (PHA) test was improved to enable the detection of antibodies to Taylorella (Haemophilus) equigenitalis in the sera of mares. Horse red blood cells (RBC) fixed with glutaraldehyde were compared with similarly treated RBC of a cow, pig and sheep for the PHA test. The horse RBC were superior to those of the other animals tested in detecting mares affected with contagious equine metritis (CEM). A PHA test using these cells as indicator and an antigen prepared from T. equigenitalis by sonication following treatment with hyaluronidase was the most satisfactory in terms of sensitivity and specificity. None of the 156 serum samples from clinically healthy mares without a history of contact with T. equigenitalis-infected stallions or mares showed PHA titers greater than 1:32 and only a few samples (7.1%) showed PHA titers of 1:32. Four of the 50 serum samples from mares affected with CEM showed PHA titers of 1:32, while most of the samples (92.0%) showed PHA titers greater than 1:32. The glutaraldehyde-fixed horse RBC sensitized with the antigen had the advantage of being reproducible for at least 7 months when preserved at 4 degrees C.

Aldehydes↗
PubMed1995

The relationship between single radial hemolysis, hemagglutination inhibition, and virus neutralization assays used to detect antibodies specific for equine influenza viruses.

Antibodies specific for equine influenza viruses are usually quantified using single radial hemolysis (SRH), hemagglutination inhibition (HI) or virus neutralization (VN). Neutralizing antibodies are thought to provide optimum protection to challenged animals. The purpose of this study was to determine the extent to which SRH and HI assays detect antibodies which neutralize equine influenza viruses. Acute and convalescent sera from 41 horses were analyzed using VN, SRH, and HI assays. These horses were present in a population of Thoroughbred racehorses during an epidemic of upper respiratory tract disease associated with influenza A/equine/Saskatoon/1/91 (H3N8), infections. Concentrations of antibodies binding to influenza A/equine/Kentucky/1/81 (H3N8), A/equine/Miami/1/63 (H3N8), and A/equine/Prague/1/56 (H7N7) were determined. Results of the VN assay were compared with results from the SRH and HI assays for acute antibody levels, changes in antibody concentrations between acute and convalescent sampling, and the occurrence of seroconversion. The correlation between assays for pre-exposure antibody levels ranged from 88% to 96%. The correlation between assays for change in antibody concentration ranged from 83% to 90% for the H3N8 viruses. This study shows that antibody concentrations specific for equine influenza virus, measured using SRH and HI assays, are highly correlated with concentrations detected using a VN assay.

Animals↗
PubMed1995

Effect of heparin on hemagglutination by Akabane and Aino viruses belonging to the Simbu group of bunyaviruses.

Heparin inhibited the hemagglutinin activity of Akabane and Aino viruses. The minimal inhibitory concentration of heparin required to inhibit 8 hemagglutination (HA) U of Akabane and Aino viruses was 10 U/ml. Goose erythrocytes failed to combine with the HA inhibitory factor of heparin. On the other hand, goose erythrocytes treated with heparinase had greatly reduced agglutinability by Akabane virus. Virus-heparin complex formation was observed by sedimenting heparin with the virus particles.

Animals↗
PubMed1995

Influence of chondroitinase on indirect hemagglutination titers and phagocytosis of Pasteurella multocida serogroups A, D and F.

Capsules of Pasteurella multocida serogroups A, D and F contain mucopolysaccharides which block antigenic determinants and prevent phagocytosis. In this study, capsules of serogroup A, D and F strains of P. multocida were depolymerized by enzyme treatment. Capsule depolymerization of serogroup D and F strains with chondroitinase increased indirect hemagglutination (IHA) test titers and enhanced phagocytosis by swine neutrophils. Capsule depolymerization of serogroup A strains with hyaluronidase increased IHA titers, but depolymerization with chondroitinase did not. When serogroup A strains were treated with a combination of chondroitinase and hyaluronidase, IHA test titers were lower than titers of the same strains treated with hyaluronidase alone. Combined enzyme treatment of serogroup D strains resulted in IHA test titers similar to those of chondroitinase treatment alone.

Animals↗
PubMedDate not supplied

Comparison of hemagglutination and hemadsorption tests for influenza detection.

Hemagglutination (HA) testing of the supernatant of primary rhesus monkey kidney cell culture tubes inoculated with clinical specimens was compared with hemadsorption (HAd) of the same tube for identifying the presence of hemagglutinin activity due to influenza. A total of 476 respiratory specimens were screened for influenza by this method. The 127 influenza or parainfluenza positive cultures showed 100% concordance between the two tests. All negative cultures were both also negative by HA and HAd. In contrast to the HAd method, HA testing was performed with less tube manipulation, less hands-on time, and less potential for cell culture tube contamination.

Animals↗
PubMed1986

The role of fimbriae of uropathogenic Escherichia coli as carriers of the adhesin involved in mannose-resistant hemagglutination.

The gene clusters encoding various P-fimbriae (F7(1), F7(2), F9 and F11) were compared. Deletion plasmids that lack the gene encoding the fimbrillin were derived from these gene clusters. Introduction of these deletion plasmids into an E. coli K12 strain resulted in non-fimbriated cells that still showed mannose-resistant hemagglutination (MRHA). However when introduced into wild type E. coli strains no MRHA was observed. Derivatives of the wild type E. coli strains with reduced amounts of O-antigen on the other hand showed MRHA when harbouring these plasmids. These results indicate that adhesion and presence of fimbriae are not necessarily linked. P-fimbriae could function as a carrier for the adhesin and thus endow adhesive capacity to cells with a complete O-antigen.

Adhesins, Escherichia coli↗
PubMed2006

Reliable serodiagnosis of imported cystic echinococcosis with a commercial indirect hemagglutination assay.

A commercially available indirect hemagglutination assay (IHA) (Echinococcosis Fumouze; Laboratoires Fumouze, Levallois-Perret, France) was evaluated using sera from 52 patients with proven cystic echinococcosis. The specificity was assessed using 247 sera from patients with various parasitic, bacterial, viral, and fungal infectious diseases; sera containing autoimmune antibodies; and sera from healthy blood donors. With a cutoff value for a positive result of 320 (as recommended by the manufacturer), the sensitivity and specificity were 88% and 98.4%; with a cutoff of 160, the sensitivity and specificity were 94% and 95.1%, respectively. The IHA is rapid, easy to perform, and is a very sensitive serodiagnostic test for cystic echinococcosis.

Animals↗