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Detection of rotavirus antibody by inhibition of reverse passive hemagglutination.

A reverse passive hemagglutination inhibition (RPHI) test was developed for detecting rotavirus antibody in patients' sera. Sheep erythrocytes coated with guinea pig antibody against Nebraska calf diarrhea virus (NCDV) were readily hemagglutinated by NCDV in a reaction called reverse passive hemagglutination (RPHA). Inhibition of this RPHA reaction was used to detect the presence of rotavirus antibody in patients' sera which cross-reacted with NCDV. The sensitivity of RPHI was consistently at least 10 times greater than that of the complement fixation test for detection of rotavirus antibody in patients' sera. Since immunoglobulin M (IgM) antibody activity detected by RPHI was destroyed by pretreatment of serum with dithiothreitol (DDT), whereas DTT-resistant antibodies such as IgG remained unchanged, the titer of IgM antibody could be distinguished from that of DTT-resistant antibody by comparing the results of two RPHI tests performed with and without DTT pretreatment. From the IgM antibody response pattern, primary and secondary rotavirus infections could be distinguished.

Adolescent↗

Detection of antibodies to Epstein-Barr virus capsid antigen by immune adherence hemagglutination.

The immune adherence hemagglutination assay was found to be as sensitive and specific as the indirect immunofluorescence technique for titration of antibodies to Epstein-Barr virus capsid antigen. Satisfactory virus capsid antigen-specific and negative control antigens for the immune adherence hemagglutination assay were prepared from cell extracts of the Epstein-Barr virus producer P3HR-1 and the Epstein-Barr virus genome-negative BJAB lymphoblastoid cell lines, respectively. As the immune adherence hemagglutination assay can be used to titrate antibodies to both the heterophil antigen of the Paul-Bunnell type and to virus capsid antigen, it offers a promising alternative to the immunofluorescence methods in the serodiagnosis of Epstein-Barr virus infections which can be performed by most diagnostic laboratories.

Antibodies, Viral↗

Heated versus unheated sera in the hemagglutination treponemal test for syphilis.

Sera (920) were tested to evaluate the use of heated versus unheated sera in the hemagglutination treponemal test for syphilis. The heated and unheated samples were tested on the same day with the hemagglutination treponemal test for syphilis kit according to the manufacturer's protocol. Agreement of results between the heated and unheated sera was 99.2%. The reading pattern of agglutination was clearer and more distinct with heated sera; therefore, based solely on our preference for the reading patterns, we suggest that heated sera be used in the hemagglutination treponemal test for syphilis.

Hemagglutination Tests↗

Comparison of different antigen preparations as substrates for use in passive hemagglutination and enzyme-linked immunosorbent assays for detection of antibody against bovine enteric coronavirus.

Purified coronavirus, detergent extracts of purified coronavirus, and virus-infected Madin-Darby bovine kidney cells were evaluated as antigen substrates in enzyme-linked immunosorbent assay (ELISA) and passive hemagglutination systems. Only detergent-extracted and -unextracted, purified viruses were reactive as antigen substrates in ELISA, whereas all three antigen preparations could be used for sensitization of erythrocytes in the passive hemagglutination assay. The passive hemagglutination system with infected cell extracts exhibited a similar level of sensitivity and specificity to the ELISA system employing purified coronavirus but enabled 300 times more tests to be performed per volume of virus-infected cell culture.

Animals↗

Evaluation of an indirect hemagglutination test for the diagnosis of human leptospirosis.

A presumptive hemagglutination test for the serological diagnosis of leptospirosis in humans is described. The antigen was prepared from a soluble alcohol extract of an andamana strain sorbed to human O-negative erythrocytes and preserved by pyruvic aldehyde fixation. In this study, the overall sensitivity of the hemagglutination test was 92% in contrast to 69% for the presumptive slide agglutination test. The specificity was 95% for the hemagglutination test in comparison with 83% for the slide test.

Agglutination Tests↗

Comparison of enzyme-linked immunosorbent assay, DNA hybridization, hemagglutination, and electron microscopy for detection of canine parvovirus infections.

Canine fecal samples were analyzed by enzyme-linked immunosorbent assays (ELISA) by using monoclonal antibodies to the canine parvovirus hemagglutinating protein. These data were compared with results obtained with DNA hybridization assays, hemagglutination assays, and electron microscopy. The highest correlation was observed between the ELISA and the hemagglutination tests, with 94.4% of samples showing agreement. Lower correlation was obtained between ELISA and DNA hybridization tests (73.3%). Correlation between ELISA and electron microscopy was 60.9%. The studies indicated that the ELISA can be used as a sensitive and specific diagnostic assay for canine parvovirus infections.

Animals↗

Comparison of indirect hemagglutination and rapid plate agglutination tests with counterimmunoelectrophoresis for typing Pasteurella haemolytica.

A rapid, simple, and accurate counterimmunoelectrophoresis (CIE) technique was developed and compared with the indirect hemagglutination and rapid plate agglutination techniques for serotyping cultures of Pasteurella haemolytica. The CIE test had 100% correlation with the conventional indirect hemagglutination test and, after serum absorption, correctly identified cultures representing the 12 established serotypes and 49 field isolates of P. haemolytica with reasonable rapidity. Cross-reactions were observed in the CIE and rapid plate agglutination tests but not in the indirect hemagglutination test with antisera prepared from the 12 established serotypes. These cross-reactions were eliminated from the CIE test but not from the rapid plate agglutination test by absorption of antisera with cells which possessed the cross-reacting antigens. Avian isolates of P. haemolytica did not type with antisera to the 12 established serotypes by any of the methods. Both homologous and heterologous reactions were observed with these strains in the rapid plate agglutination and CIE tests with antisera prepared from six selected cultures. These results support the previous finding that the taxonomic relationship of these avian strains to P. haemolytica is questionable.

Agglutination Tests↗

Comparison of enzyme-linked immunosorbent assay, hemagglutination inhibition, and passive latex agglutination for determination of rubella immune status.

Hemagglutination inhibition, enzyme-linked immunosorbent assay (ELISA), and passive latex agglutination were used to test 495 human serum samples for determination of rubella immunity. Overall agreements for immune status were as follows: hemagglutination inhibition versus ELISA, 94.7% (469 of 495); hemagglutination inhibition versus passive latex agglutination (1:10 dilution), 99.2% (491 of 495); and passive latex agglutination (1:10 dilution) versus ELISA, 94.7% (469 of 495). Both ELISA and passive latex agglutination are satisfactory for rubella immunity screening, with the reservation that the ELISA method examined yielded a large number of false-negative results.

Adult↗

Comparison of a simple latex agglutination test with hemolysis-in-gel, hemagglutination inhibition, and radioimmunoassay for detection of rubella virus antibodies.

Rubella virus antibodies were measured in 300 sera from pregnant women visiting a maternity center by using a new, simple latex test, Rubalex. The results were compared with those obtained by using hemolysis in gel, hemagglutination inhibition, and radioimmunoassay. The sensitivity of the latex test was 100, 98.0, and 99.6% when compared with hemolysis in gel, hemagglutination inhibition, and radioimmunoassay, respectively. Its comparative specificity was 96.2, 95.7, and 90.7%, and the predictive value of a positive result was 99.2, 99.2, and 98%, respectively. When assayed with the British standard anti-rubella serum its sensitivity was 11 IU/ml. The latex test gave a positive result within 2 min, and 87% of the positive samples had already reacted after 1 min. The negative results remained as such for at least 8 min. No prozone effect was observed for sera with hemagglutination inhibition titers from 256 to 2,048. We concluded that the latex test, Rubalex, was readily applicable for measuring rubella immunity with a reaction time of 2 min in undiluted samples.

Antibodies, Viral↗

Serodiagnosis by passive hemagglutination test and verotoxin enzyme-linked immunosorbent assay of toxin-producing Escherichia coli infections in patients with hemolytic-uremic syndrome.

Eight cases of hemolytic-uremic syndrome in which no pathogens were isolated were diagnosed serologically by a passive hemagglutination assay and a verotoxin (VT; Shiga-like toxin) enzyme-linked immunosorbent assay (ELISA). The passive hemagglutination assay employed formalinized sheep erythrocytes sensitized with soluble native antigen or heat-treated antigen (lipopolysaccharide [LPS]) from Escherichia coli O26, O111, O128, and O157 or flagellar antigen of nine different H serogroups of E. coli: H2, H7, H8, H10, H11, H12, H18, H19, and H25. All patients had antibodies against the native antigen and/or the LPS of E. coli O157, but positive agglutination with H7 was observed only in one patient. In the VT-ELISA with plates coated with purified VT 1 or VT 2, antibody against VT 2 was observed in the sera of five of six patients examined, but none of the patients possessed VT 1 antibody. These results indicate that the causative pathogen in these eight hemolytic-uremic syndrome cases is likely to be VT-producing E. coli O157. The passive hemagglutination assay described here is a very sensitive, simple, and rapid method. This assay is highly recommended for the serological diagnosis of VT-producing E. coli infections, particularly in patients infected by serogroup O157 strains. Furthermore, the VT-ELISA is useful in studying the role of VT in hemolytic-uremic syndrome.

Animals↗

Whole-blood hemagglutination inhibition test for venereal disease research laboratory (VDRL) antibodies.

Nontreponemal antibody tests such as the Venereal Disease Research Laboratory (VDRL) test are carried out on serum and widely used as screening tests for syphilis. The aim of the present study was to develop a screening test for syphilis making use of whole blood and VDRL liposomes. Antibody to human red blood cells was conjugated to VDRL liposomes and reacted with a diluted sample of patient whole blood. A total of 951 samples were tested by the new test and the VDRL tube test. All 49 VDRL samples positive by the VDRL test showed inhibition of hemagglutination in the whole-blood test (sensitivity, 100%). Of 902 samples with negative results by the VDRL test, 901 caused hemagglutination when tested with the liposomes (specificity, 99.9%). The hemagglutination inhibition method tests for syphilis in a simple one-step procedure in which whole blood is added to a tube containing liposomes. The new test has potential for point-of-care testing in developing countries.

Animals↗

Comparison of indirect hemagglutination and indirect immunofluorescence tests with microneutralization tests for detection of type-specific Herpesvirus hominis antibody.

Indirect hemagglutinating and immunofluorescent antibody responses to Herpesvirus hominis types 1 and 2 were compared to neutralizing antibody responses in infected humans from whom H. hominis type 1 or 2 was isolated. The indirect immunofluorescent antibody test was shown to be the most sensitive and specific for primary human infections. The sensitivity and specificity of the indirect hemagglutination and the immunofluorescent antibody tests were shown to be equal to that of the microneutralization test among patients who had primary or recurrent H. hominis type 2 infections. It is suggested that the indirect hemagglutination test is preferable for assaying large populations for previous infection with H. hominis type 2 because it is rapid, easier to perform, and more economical. The intermediate range of titer differences (deltat) between H. hominis types 1 and 2 previously reported to be due to infections with both viruses was shown to occur in all three tests among patients with primary infections with either virus.

Antibodies, Viral↗

Adeno-associated virus serotype 4 (AAV4) and AAV5 both require sialic acid binding for hemagglutination and efficient transduction but differ in sialic acid linkage specificity.

Adeno-associated virus serotype 4 (AAV4) and AAV5 have different tropisms compared to AAV2 and to each other. We recently reported that alpha 2--3 sialic acid is required for AAV5 binding and transduction. In this study, we characterized AAV4 binding and transduction and found it also binds sialic acid, but the specificity is significantly different from AAV5. AAV4 can hemagglutinate red blood cells from several species, whereas AAV5 hemagglutinates only rhesus monkey red blood cells. Treatment of red blood cells with trypsin inhibited hemagglutination for both AAV4 and AAV5, suggesting that the agglutinin is a protein. Treatment of Cos and red blood cells with neuraminidases also indicated that AAV4 bound alpha 2--3 sialic acid. However, resialylation experiments with neuraminidase-treated red blood cells demonstrated that AAV4 binding required alpha 2--3 O-linked sialic acid, whereas AAV5 required N-linked sialic acid. Similarly, resialylation of sialic acid-deficient CHO cells supported this same conclusion. The difference in linkage specificity for AAV4 and AAV5 was confirmed by binding and transduction experiments with cells incubated with either N-linked or O-linked inhibitors of glycosylation. Furthermore, AAV4 transduction was only blocked with soluble alpha 2-3 sialic acid, whereas AAV5 could be blocked with either alpha 2--3 or alpha 2-6 sialic acid. These results suggest that AAV4 and AAV5 require different sialic acid-containing glycoproteins for binding and transduction of target cells and they further explain the different tropism of AAV4 and AAV5.

Animals↗

Hemagglutination by murine hepatitis viruses. Absence of detectable activity in strains 3, A59, and S grown on DBT cells.

Erythrocytes from twelve mammalian and avian sources in ten different buffers at three incubation temperatures could not be hemagglutinated with murine hepatitis virus (MHV) strains 3, A59, or S grown on DBT cells. Viral antigen preparation in the absence of fetal calf serum, partial virus purification, or various concentrations of red blood cells still failed to yield detectable hemagglutinating activity. Thus, the newly described MHV-DVIM remains the only hemagglutinating strain of murine coronavirus.

Animals↗

Counterimmunoelectrophoresis using an arc 5 antigen for the rapid diagnosis of hydatidosis and comparison with the indirect hemagglutination test.

A modified counterimmunoelectrophoresis and a conventional indirect hemagglutination test were compared for routine diagnosis of human hydatid disease in an endemic area in the Middle East. Counterimmunoelectrophoresis was performed on a cellulose acetate membrane with dilutions of a commercially available antigen which interacts with sera of patients with confirmed hydatid disease to produce the arc 5 precipitin line. The test was performed with unconcentrated human sera and the lines stained in an aqueous solution of Ponceau red. Sensitivity (95.5% vs. 93.2%) and specificity (99.2% vs. 89.9%) were higher with counterimmunoelectrophoresis than with indirect hemagglutination. Cross-reactivity with sera of patients with other parasitic infections was noted with indirect hemagglutination but not with counterimmunoelectrophoresis. There was no cross-reactivity with sera of patients with autoimmune disorders by either test.

Antibodies↗

[Use of the indirect hemagglutination reaction for detecting antibodies to Pseudomonas pyocyanea in acute suppurative destructive pneumonia].

No less than 4-fold increase in the antibody titres to Pseudomonas aeruginosa during the infectious process served as laboratory confirmation of its participation in the infectious process. In 49 of 91 patients hemagglutining titre exceeded the diagnostic one (1:640). In 9 patients (chiefly in infants) hemagglutinin titre remained low, but there was a rise of antibody level to Pseudomonas aeruginosa during the disease. High hemagglutinin titres were noted in 12 patients on admission to the clinic, with reduction of the antibody titres at the late periods of the disease. Antibody titres remained unchanged during the disease in 15 patients. In 3 cases the indirect hemagglutination test was assesed as negative. In the rest of the patients hemagglutinin titres varied within the range of the diagnostic titre. Thus, the indirect hemagglutination test with erythrocytic diagnostic agent permitting to determine antibodies to Pseudomonas aeruginosa could be used at the clinic in combination with bacteriological and other investigations for establishing etiology of the destructive process.

Acute Disease↗

[Study of arenaviruses in the indirect hemagglutination reaction].

The indirect hemagglutination test with erythrocytes sensitized with antibody to LCM and Tacaribe viruses, most typical members of of arenaviruses, has been developed. This test is a sensitive method for detection of arenaviruses in infected materials in vitro. The indirect hemagglutination test and the indirect hemagglutination-inhibition test revealed cross-interaction between LCM and Tacaribe viruses which opens new approaches for group and species identification of arenaviruses.

Hemagglutination Tests↗

[Comparison of characterization of hemagglutination and HA1 sequence of currently circulating strains of influenza A (H3N2) virus in Zhejiang province and in Japan].

OBJECTIVE: To compare the characterization of hemagglutination and the sequence of HA1 region of the influenza A (H3N2) virus in Zhejiang province and Japan in recent years. METHODS: Hemagglutinin antigenic analysis of influenza A (H3N2) viruses which were isolated in Zhejiang province was carried out and to compare their characterization of hemagglutination and the sequence of amino acids in HA1 region with the virus strains from Japan during the same period. RESULTS: The antigenicity and the amino acid sequence in HA1 region of the virus strains prevalent in Zhejiang in recent years have changed to some extent. Most of the virus strains were lack of hemagglutination reaction to chicken red blood cell and became "O" phase virus strains. The amino acid sequence of HA1 region of the virus strains exhibited only 94.82% of its homology with that of A/Wuhan/359/95 strain. The virus strains of Japan in the same period showed similar characteristics. CONCLUSION: HA1 gene variation of influenza A (H3N2) viruses was the main cause leading to the spread of influenza in the last years.

Amino Acid Sequence↗