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Antibody responses induced by immunization with a Japanese rabies vaccine determined by neutralization test and enzyme-linked immunosorbent assay.

The immunogenicity of a Japanese purified chick embryo cell culture rabies vaccine (PCECV) was examined. Serum samples were obtained from 86 subjects after pre-exposure or post-exposure prophylaxis. Rabies antibody titres were determined by neutralization test and enzyme-linked immunosorbent assay (ELISA). Titres higher than 0.5 international units (IU)/ml were demonstrated by neutralization test in all the 19 subjects after three-time pre-exposure immunization on days 0, 30 and 180. Titres higher than 0.5IU/ml were also demonstrated by neutralization test in all the 23 subjects after four-time post-exposure immunization on days 0, 3, 7 and 14. There was a correlation between neutralization and ELISA antibody titres (r=0.697); however, neutralization titers were higher than ELISA titres for most of the samples. The results suggest that current Japanese rabies vaccine induces recommended levels of neutralizing antibodies after pre- and post-exposure prophylaxes.

Antibodies, Viral↗

Modified Hudson-Kreitman-Aguade test and two-dimensional evaluation of neutrality tests.

There are a number of polymorphism-based statistical tests of neutrality, but most of them focus on either the amount or the pattern of polymorphism. In this article, a new test called the two-dimensional (2D) test is developed. This test evaluates a pair of summary statistics in a two-dimensional field. One statistic should summarize the pattern of polymorphism, while the other could be a measure of the level of polymorphism. For the latter summary statistic, the polymorphism-divergence ratio is used following the idea of the Hudson-Kreitman-Aguadé (HKA) test. To incorporate the HKA test in the 2D test, a summary statistic-based version of the HKA test is developed such that the polymorphism-divergence ratio at a particular region of interest is examined if it is consistent with the average of those in other independent regions.

Computer Simulation↗

A rapid plaque neutralization test for bluetongue virus.

A bluetongue virus plaque neutralization test, done in small tissue culture wells (2.0 cm2) using minimal antibody-virus contact time, was found to be of comparable sensitivity to more classical methods involving preincubation. By using the cell culture plates as serum dilution wells and adding cells directly to the virus-antibody mixture, a considerable saving in time is realized. Because of the small size of the wells, reagent cost and incubator space requirements are reduced making the test much more attractive for wide scale use, either as a screening test or one that yields dose-response lines.

Animals↗

The surface antigens of orthopoxviruses detected by cross-neutralization tests on cross-absorbed antisera.

Cross-neutralization tests were done on accepted species and recently isolated members of the genus Orthopoxvirus using antisera which had been separately absorbed with the various viruses. The results provided evidence for the involvement of four neutralizing antigens, and their distribution among 13 virus strains was determined. Monkeypox (Congo-8-Lombe), camelpox (Gorgan), ectromelia (Mill Hill), 'Lenny' and elephant poxviruses had distinctive antigenic formulae. Lister and Wyeth vaccines were indistinguishable but different from Copenhagen and EM63 vaccines which were themselves distinct. Cowpox (Brighton), buffalopox (BP4), MK 10, and Moscow poxviruses were indistinguishable. Examples were found where viruses shared surface antigens but were not all neutralized by antibody to them. this reduced the practical value of the technique for virus identification. Evidence was also obtained for the existence in some viruses of a fifth antigen, antibody to which could block neutralization by antibody to one particular antigen.

Antigens, Surface↗

Immunity to a 3-methylcholanthrene-induced fibrosarcoma in the C57BL/6 mouse: in vivo analysis by the adoptive tumor neutralization test.

Analyses of the adoptive tumor neutralization test (modified Winn test) in C57BL/6 mice made immune to a 3-methylcholantheene-induced fibrosarcoma showed that the reaction was mediated by a thymus-derived lymphocyte, it was tumor-specific, and the resistance of the immunized donor mouse to the challenge was strongly correlated with the protection of the recipient mouse. Proliferation of immune cells and close contact between tumor cells and immune T-cells were required. The hypothesis of a participation of the recipient in the reaction was considered because of the lack of adoptive protection of pangenic nude mice.

Animals↗

Rapid focus reduction neutralization test of influenza A and B viruses in microtiter system.

A rapid neutralization test for influenza A and B viruses was developed. In this method, a 96-well tissue culture plate was used for the preparation of cell monolayers and the peroxidase-antiperoxidase staining technique was used for the visualization of foci infected with these viruses. In the presence of trypsin and tragacanth gum, clear foci developed 1 day after infection. A linear relationship between virus dilutions and numbers of foci was observed. When neutralizing antibodies in some test sera were assayed, a good correlation was observed between the titers obtained by the focus method and those obtained by the ordinary plaque method. In addition, many serum specimens were investigated by the neutralization test, and it was demonstrated that the test is useful for serological studies of influenza.

Animals↗

Hemolysis-in-gel and neutralization tests for determination of antibodies to mumps virus.

A hemolysis-in-gel test for the demonstration of antibodies to mumps virus is described. The results were compared with those of neutralization tests using a modified microtechnique. In the neutralization test viral replication was demonstrated by the hemadsorption of guinea pig erythrocytes, the visibility of which could be further enhanced by the use of o-tolidine. Good correlation was found between the results of the two techniques. The hemolysis-in-gel test was simple to perform, rapid, sensitive, and shown to be a useful test for the demonstration of mumps antibodies.

Antibodies, Viral↗

Assessment of IgG antibodies against yellow fever virus after vaccination with 17D by different assays: neutralization test, haemagglutination inhibition test, immunofluorescence assay and ELISA.

We analysed serum samples of 209 subjects immunized with yellow fever vaccine 17D by different assays: neutralization test, immunofluorescence assay, haemagglutination inhibition test and ELISA, for presence of 17D-specific antibodies. Serum samples were taken from a few weeks up to 35 years after vaccination. The neutralization test had the highest sensitivity. There was no correlation of results between the serological assays. Considering NT titres > 1:10 as indicating protection, we found that about 75% of subjects remained immune even 10 years after vaccination, with a median NT titre of 1:40 in reactive sera.

Adolescent↗

Microculture plaque neutralization test for California group arboviruses.

A microculture plaque neutralization test is described for California-group arboviruses that is as precise and quantitative as the standard test conducted in 60-mm petri dishes. It was shown that there was no significant between-panel or between-day variation in determinations and that a single pipette could be used for all serum-dilution levels within a titration without inoculum carry-over effect. The experimental protocol and statistical methods used produce 50% neutralization end points that meet the assumptions of parametric statistics. This permits the power and versatility of the analysis of variance to be exploited in testing for treatment effects in serological and immunological studies with viruses.

Analysis of Variance↗

[Epizoologic studies of the detection of antibodies against Aujeszky's disease virus in sera and blood eluates of swine from Thailand using ELISA ("Enzygnost," Behring), serum neutralization test and "Aujeszky Latex Kit" (Iffa Merieux)].

The results of three tests for Aujeszky's disease were analysed and compared. The presence of Aujeszky's antibodies was determined by "Enzyme-linked-Immunosorbent-Assays" (ELISA, "Enzygnost"), Behring company, Marburg; "Serum-Neutralization-Tests" (SNT); and "Latex Agglutination-Tests" (LT, "Aujeszky-Latex-Kit"), Iffa Merieux company, Laupheim. Whole blood and sera samples were taken from 805 swine from 26 of Thailand's provinces. These samples were analysed to determine if eluates of whole blood on filter paper were as effective as corresponding sera samples in determining the presence of Aujeszky's disease antibodies. From a total of 805 samples, 26% of the serum and 18% of the blood eluate samples showed a positive result when tested by the ELISA method. Clearly, testing whole blood eluates provides results which are inferior to results from sera samples. Therefore the ELISA whole blood eluates test can only be recommended with reservations. Further testing was done on 645 serum samples using SNT. Samples tested were those which gave negative, suspicious, or weakly positive results when tested by ELISA. Using SNT, 23% of these showed a positive result. Many serum and blood eluate samples were also tested by LT. Most of these test samples were chosen because they were deemed suspicious. Suspicious samples were defined as those which had deviant test results. According to these results the sensitivity of LT was between the sensitivity of SNT and ELISA. Owner survey responses tended to state that few animals had been vaccinated. This coupled with the frequency of antibody occurrence proves the high rate of infection among Thailand's swine population.

Animals↗

Detection of postvaccination mumps virus antibody by neutralization test, enzyme-linked immunosorbent assay and sensitive hemagglutination inhibition test.

A group of 251 children aged 2-3 years given live attenuated mumps virus vaccine PAVIVAC of Czechoslovak production were tested for antiparotitis antibody levels in pre- and postvaccination sera by neutralization test (NT), enzyme-linked immunosorbent assay (ELISA) and sensitive hemagglutination inhibition test, enhanced by heterologous antibody to human immunoglobulin G (E-HIT). The prevaccination findings were as follows: positive ELISA IgG titres, neutralization antibodies and hemagglutination inhibition antibodies were present in, respectively, 35%, 25.9% and 27.9% of the sera. Postvaccination seroconversions were evaluated in 159 susceptible vaccinees whose prevaccination sera had been negative by all three tests. The lowest seroconversion was detected by NT (74.2%), seroconversions by ELISA and E-HIT were appreciably higher (82.4% and 86.8%, respectively). The seven children showing a seroconversion by E-HIT but not by ELISA had a 4 fold increase of anti-mumps ELISA IgG antibodies as well, but the rise of antibody titres was at a level falling in the range below the positivity criterion for ELISA. The statistically evaluated detection rate for antibodies was significantly higher (significance test "t") by ELISA as compared with neutralization test. However, antibody levels (geometric mean titres) were 8-10 times lower in postvaccination sera than in convalescent sera of 30 children with mumps in all three tests.

Antibodies, Viral↗

The antiglobulin neutralization test: a procedure for evaluating the efficacy of washing red cells for antibody elution.

The antiglobulin neutralization test, a procedure designed to detect neutralizing concentrations of IgG, is compared with the sulfosalicylic acid test and the indirect antiglobulin test as a means of evaluating efficacy of protein removal by washing red cells prior to antibody elution. Variables that may affect the results of the antiglobulin neutralization test and the sulfosalicylic acid test are discussed.

Journal Article↗

Arbovirus neutralization tests with Peruvian sera in Vero cell cultures.

Selected human sera from Peru, previously examined by the haemagglutination-inhibition (HI) test with a number of arboviruses, were reexamined by neutralization tests carried out in Vero cell cultures. Results confirmed and extended the HI findings, indicating that the antibodies detected were evoked by Eastern equine encephalitis, Mayaro, Venezuelan equine encephalitis, Ilheus, St Louis encephalitis, yellow fever, Caraparu, and Guaroa viruses.

Antibodies↗

Estimation of seroprevalence of encephalomyocarditis in Dutch sow herds using the virus neutralization test.

Encephalomyocarditis virus (EMCV) has been found on pig farms worldwide and can cause myocarditis in young pigs and reproduction disorders in sows. So far, clinical signs of EMCV have not been reported in the Netherlands. The aim of this study was to estimate the seroprevalence of EMCV infection in Dutch sow herds. A total of 277 Dutch sow herds were randomly selected, from which 3237 serum samples were collected. These samples were tested for EMCV antibodies using the virus neutralization test (VN test). The apparent prevalence of EMCV antibodies was 9.3% in the total sow population, and the apparent herd prevalence was 58.8%. An exact determination of the prevalence of EMCV infections in the Dutch sow population was not possible because the characteristics of the VN test under field circumstances were not known. However, Dutch sow herds seem to be infected with EMCV because the distribution of positive blood samples in the tested sow population was significantly different from that expected if random false-positive reactions had occurred.

Animals↗

A refined complement-enhanced neutralization test for detecting antibodies to Junin virus.

A refined, complement-enhanced, plaque-reduction neutralization test was developed for measuring neutralizing antibodies against Junin (Argentine hemorrhagic fever) virus. The assay measured neutralizing antibodies after natural as well as vaccine-induced Junin virus infections. Among vaccinated individuals, titers were 2-4-fold higher than those obtained with conventional assays, without loss of specificity. Enhanced sensitivity was achieved by using a standardized complement source (vs human or animal serum) for virus dilution, incubation of virus-serum mixtures at 36 degrees C for 2 h (vs overnight at 4 degrees C) prior to plaque assay, control of age and density of cell monolayers, and variation in overlay conditions.

Analysis of Variance↗

[Use of the neutralization test for mass population studies].

The authors applied the antibody neutralization test (ANT) for detection of causative agent of intestinal infections in feces of healthy population in Turkmenia. Some part of the material was studied in parallel with the aid of the ANT and by the bacteriological method. As a result of comparison it appeared that in using the ANT causative agents of intestinal infections were revealed much more frequently than by the bacteriological method; in this connection this serological method can be recommended as an auxiliary one for the diagnosis of intestinal infections.

Diagnosis, Differential↗