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PubMed1980

A micro-neutralization test for flavivirus antibodies.

A micro-neutralization test for the detection of flavivirus antibodies is described. The test utilizes the PS line of cells in flat bottomed microtiter plates to detect the neutralization of cytopathic effect by specific antibodies. Cross-neutralization of flaviviruses found in Southeast Asia with homologous and heterologous hyperimmune mouse ascitic fluids revealed the test to be specific. The test is simple and inexpensive to perform.

Antibodies, Viral↗
PubMed1985

Micro-neutralization test for mumps virus using the 96-well tissue culture plate and PAP (peroxidase-antiperoxidase) staining technique.

A rapid micro-test method for mumps virus neutralization was developed. In this method, a 96-well tissue culture plate was used for preparation of cell monolayers and the PAP staining technique was used for visualization of mumps virus infected cells. Clusters of infected cells were observed as a focus and the numbers of foci could be counted by the naked eye 2 days after the infection. A linear relationship between virus dilutions and focus numbers was observed. When neutralizing antibodies in sera from cases of natural mumps infection were assayed, a good correlation was observed between those obtained by the focus reduction method applying the micromethod and those obtained by the ordinary plaque method. Our results indicate that this micromethod is useful in mumps virus neutralization tests and it has many advantages over other methods previously reported.

Animals↗
PubMed1978

[Detection of antibodies to Rickketsia prowazekii by using the antigen neutralization test].

The authors studied a possibility of using the antigen neutralization test with dry immunoglobulin typhus erythrocytic diagnostic agent for the purpose of detection of Rickettsia prowazeki antibodies. Blood sera of 315 healthy persons, 24 patients with sporadic typhus, and 18 laboratory animals immunized with R. sibirica and R. burneti, as well as with Proteus OX19 were examined. The results obtained pointed to the high specificity and sensitivity of the given serological test. A possibility of its use for antibody detection both in the typhus patients and in persons who sustained this infection in the past was demonstrated. In difference from the complement fixation test it permits to study anticomplementary sera.

Antibodies, Bacterial↗
PubMed1981

Role of virus strain in conventional and enhanced measles plaque neutralization test.

A low-passage Edmonston strain of measles virus gave on the average four-fold higher antibody titers than two high-passage strains of Edmonston virus when used in a virus plaque neutralization test. Heterologous anti-human immunoglobulin did not affect antibody titers obtained with the low-passage virus but enhanced antibody titers obtained with the high-passage viruses on the average eight-fold. These differences in virus sensitivity to neutralization and neutralization potentiation by anti-immunoglobulin were interpreted as being possibly due to differences in density or accessibility of the functional antigens of measles virus. The conventional plaque neutralization test utilizing the low-passage Edmonston virus was about 10 times more sensitive than the virus CPE-neutralization test, 60 times more sensitive than the measles hemagglutination-inhibition test and 220 times more sensitive than the measles complement-fixation antibody test.

Animals↗
PubMed1990

Serodiagnosis of canine herpesvirus infection--development of an enzyme-linked immunosorbent assay and its comparison with two improved methods of serum neutralization test.

An enzyme-linked immunosorbent assay (ELISA) for the diagnosis of canine herpesvirus (CHV) infection using antigen prepared by solubilizing infected cells was developed. The ELISA and two improved methods of serum neutralization test, the microplate serum neutralization test (MSNT) with complement and the 50% plaque reduction (PR) assay with complement, were compared for the results of antibody detection from a total of 557 field canine sera. Of 529 sample sera that were negative in the MSNT with complement, 119 were ELISA positive, and this result together with time course of serum antibody detection in a dog experimentally infected with CHV strongly suggested that the MSNT with complement is less sensitive for the detection of antibody in CHV infected dogs, especially those in early stages of infection. A correlation was found between the titers measured by the ELISA and 50% PR assay with complement, however, for field use, the ELISA is recommended as a highly sensitive test method of serodiagnosis of CHV infection adequate for dealing with a large number of samples with less demand on time and effort.

Animals↗
PubMed1979

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↗
PubMed1982

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↗
PubMed1979

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↗
PubMed1990

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↗
PubMed1976

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↗
PubMed1974

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↗
PubMed1990

[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↗
PubMed1987

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↗
PubMed1972

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↗
PubMed1990

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↗
PubMed1976

[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↗
PubMedDate not supplied

Comparison of indirect immunofluorescence test for measles antibodies with haemagglutination inhibition and plaque neutralization tests.

Indirect Immunofluorescence (IFA), Plaque Reduction Neutralization (PRN) and Haemagglutination Inhibition (HI) tests for measles antibodies were carried out in 197 sera obtained from umbilical cord and vaccinated children. The IFA was also applied to blood samples collected with filter paper. IFA results demonstrated that the test is relatively simple to perform, with good reproducibility for different antigen lots. Good correlation was obtained between IFA, PRN and HI antibody titers. Better correlation was demonstrated with IFA and PRN than with HI and PRN tests. Sensitivity of IFA in detecting antibody was less effective than PRN, however more effective than HI using rhesus monkey red blood cells. PRN antibody titers over 100 were detected by IFA but not by HI (9.7% with negative results). IFA may be of considerable practical use and able to substitute HI in seroepidemiological surveys and to evaluate vaccine efficacy. It also can be simplified by employing filter paper collected samples.

Adult↗