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Screening of pig sera for antibodies to Japanese encephalitis virus using a dot enzyme immunoassay and IgM capture ELISA: comparison with the hemagglutination inhibition and plaque reduction neutralization tests.

A dot enzyme immunoassay for determination of antibodies to Japanese encephalitis virus was designed for use as a field technique for the surveillance of Japanese encephalitis virus activity among domestic pigs. The test was compared with the neutralization test and the hemagglutination inhibition test and found to be more sensitive than the hemagglutination inhibition test and comparable to the neutralization test in sensitivity but more simple to perform than either the neutralization or the hemagglutination inhibition tests. An IgM capture ELISA for the determination of JEV specific porcine IgM was also utilized to determine current infection rates in pigs. The tests which do not involve the determination of specific IgM are better used for testing sentinel animals for providing clues as to the rate of transmission of JEV among pigs. IgM tests determining acute infection are less likely to be useful unless animals are tested very frequently or if a great number of animals are tested at any one time.

Animals↗

Neutrality tests using DNA polymorphism from multiple samples.

The polymorphism of a gene or a locus is studied with increasing frequency by multiple laboratories or the same group at different times. Such practice results in polymorphism being revealed by different samples at different regions of the locus. Tests of neutrality have been widely conducted for polymorphism data but commonly used statistical tests cannot be applied directly to such data. This article provides a procedure to conduct a neutrality test and details are given for two commonly used tests. Applying the two new tests to the chemokine-receptor gene (CCR5) in humans, we found that the hypothesis that all mutations are selectively neutral cannot explain the observed pattern of DNA polymorphism.

Chromosome Mapping↗

ELISA as a possible alternative to the neutralization test for evaluating the immune response to poliovirus vaccines.

This review describes several enzyme-linked immunosorbent assay (ELISA) techniques proposed to replace the neutralization test for detecting neutralization-relevant antibodies to polioviruses in recipients of inactivated poliovirus vaccine and oral poliovirus vaccine, and for seroepidemiologic studies. Comparisons of results from ELISA and the neutralization test suggest that ELISA variants, based on the principle of blocking or binding inhibition that emulate the neutralization test, might offer an alternative to the neutralization test. However, to replace the neutralization test with ELISA would first require extensive studies with very large numbers of serum samples, including sera having low titers of neutralizing antibodies, in order to obtain reliable and statistically sound validation.

Animals↗

Transmissible gastroenteritis virus: plaques and a plaque neutralization test.

A plaquing system and plaque neutralization test in porcine thyroid cells were used to study different transmissible gastroenteritis isolates and hemagglutinating encephalomyelitis virus. Among transmissible gastroenteritis virus isolates, plaque size varied considerably and mixed size ranges sometimes occurred. The most recently isolated viruses produced smaller plaques than the laboratory viruses or hemagglutinating encephalomyelitis virus. All transmissible gastroenteritis virus isolates reacted in the plaque neutralization test with a transmissible gastroenteritis virus antiserum which showed no activity against hemagglutinating encephalomyelitis virus. Plaque neutralization results both from experimentally infected pigs and following a field outbreak demonstrated the reliability of this test and its greater sensitivity than the conventional tube test.

Animals↗

Reproducibility of a virus-neutralization test for infectious laryngotracheitis virus.

A virus-neutralization test for infectious laryngotracheitis virus was performed in microtiter plates using standard techniques. To assess the reproducibility of the test, 11 sera were each titrated repeatedly once a week for 8 weeks, and the results were compared to a standard. The standard used for reproducibility was that the 95% logarithmic confidence intervals of the mean calculated from three titrations of the same serum had to be smaller than the logarithmic distance "within" two microtiter plate wells. For the virus neutralization test to give reproducible results, such confidence intervals had to fall "within" two wells at least 75% of the time. Over the 8 weeks, percent reproducibility varied from 43.5% to 81.5%. The infectious laryngotracheitis virus-neutralization test did not meet our defined standard of reproducibility with positive antisera. Results with negative control sera were reproducible, however. Percent reproducibility varied from 31.8% to 93.8% for different sera tested, but it was not related to the titer of the sera.

Animals↗

Plaque reduction neutralization test for human cytomegalovirus based upon enhanced uptake of neutral red by virus-infected cells.

Foci of cells infected with human cytomegalovirus were noted to stain more intensely than uninfected cells with neutral red, and this provided the basis for development of a plaque assay and plaque reduction neutralization test for cytomegalovirus. Plaques demonstrable by neutral red staining could be counted at 8 days after infection; thus, results could be obtained earlier than for plaque assay systems based upon the viral cytopathic effect, a fewer manipulations were required for staining cell monolayers to demonstrate plaques. Certain variables affecting plaque size and numbers and antibody titers were defined. Addition of fresh guinea pig complement to the reaction mixtures markedly enhanced cytomegalovirus-neutralizing antibody titers of hyperimmune animal sera, but titers of human sera were enhanced only two-or fourfold.

Animals↗

[Vaccination of ostriches (Struthio camelus, Linnaeus, 1758) against Newcastle disease: evidence for vaccine compatibility and seroconversion after vaccinations using the hemagglutination inhibition and virus neutralization tests].

Newcastle disease (ND) remains to be the worldwide most important infectious disease of poultry. This epizootic is in Germany and many other countries a notifiable disease. Prophylactic vaccination is the major tool for the control of ND in poultry and other birds. Eighty-three ostriches (Struthio camelus) which were kept on farms in Germany were checked for the presence of NDV-specific antibodies. Some of these birds are said to be vaccinated against Newcastle disease. Only some of these ostriches contained antibodies which were measurable in haemagglutination inhibition and virus neutralisation tests. Twenty-three previously unvaccinated ostriches were vaccinated with commercially available vaccines. Both the LaSota live and inactivated oil emulsion vaccines were well tolerated following conjunctival or subcutaneous application, respectively. Neither local nor systemic side reactions were observed. After the vaccinations high antibody titres were detected in hemagglutination inhibition and virus neutralisation tests. A strong correlation between both established methods (r = 0.92; < 0.001) were noted.

Animals↗

Potency control of diphtheria component in adsorbed vaccines by in vitro neutralization tests.

Samples from 20 lots of dT vaccine and from 20 lots of DTP vaccine were used to standardize and validate the Vero cell and the toxin binding inhibition (ToBI) tests for the potency control of diphtheria component. For the Vero cell method, violet crystal solution was used to stain the cells and estimate the endpoint of diluted diphtheria antitoxin. Diphtheria anatoxin was used for performing the ToBI test instead of toxin. The results obtained by both in vitro tests were similar to those obtained by in vivo toxin neutralization test in guinea pigs. The various analysis and the chi(2) test applied to evaluate the reproducibility and homogeneity, respectively, among in vitro tests and in vivo toxin neutralization test did not detect statistical significant difference for both analysed vaccines. An excellent correlation among in vitro tests and in vivo neutralization test was observed by Spearman's correlation coefficient.

Adsorption↗

Comparison of the latex agglutination test with the hemagglutination inhibition test, enzyme-linked immunosorbent assay, and neutralization test for detection of antibodies to rubella virus.

The ability of a rapid, latex agglutination test to diagnose rubella infection and to measure immune status was evaluated by comparison with the hemagglutination-inhibition (HAI) test, enzyme-linked immunosorbent assay (ELISA), and the neutralization (NT) test. The latex agglutination test accurately detected serological conversions in 74 pairs of sera representing 21 natural infections and 53 immunizations. The antibody levels of 276 sera from the general population were determined by latex agglutination, HAI, and ELISA. The correlation coefficients between the titers obtained by HAI and latex agglutination and by ELISA and latex agglutination were statistically significant. Results on 12 sera did not agree when measured by the three tests. These sera were included among the 196 specimens tested by NT. The correlation coefficient between NT and latex agglutination titers was statistically significant. There was one serum positive by latex agglutination but negative by NT, and five sera were negative by latex agglutination but had titers of 4 to 8 in the NT. The relative sensitivity of detecting antibody was greater by latex agglutination than by HAI. An additional 49 sera containing residual nonspecific hemagglutinin inhibitors were evaluated by latex agglutination and NT. The untreated sera showed no false positive reactions, and 36 of 39 NT positive sera were positive in the latex agglutination test.

Antibodies, Viral↗

Effect of misoriented sites on neutrality tests with outgroup.

Several neutrality tests use outgroups to infer the ancestral and derived states for polymorphism data. However, homoplasy can result in the incorrect inference of the derived variant. We show that empirically derived rates of misorientation strongly influence Fay and Wu's H-test, especially when the sample size is large.

Mutation↗

Inosithin neutralization test to measure lupus anticoagulants.

The inosithin neutralization test was performed in 14 patients in whom lupus anticoagulant was detected. To test its specificity, it was also performed in 10 patients with severe hemophilia and in three patients with Factor VIII inhibitors. Prolonged kaolin clotting time was corrected by adding varying amounts of inosithin (Asolectin, 0.19 to 100 micrograms), a soybean-derived phospholipid, in all patients with lupus anticoagulant but not in patients with hemophilia or in two patients with Factor VIII inhibitors. In one patient, both Factor VIII inhibitors and lupus anticoagulant were present. The concentration of lupus anticoagulant in a patient was expressed as the amount of inosithin (measured in micrograms) required to normalize the prolonged kaolin clotting time. This amount correlated significantly with the occurrence of thrombosis and/or recurrent abortion. The inosithin neutralization test is useful to measure lupus anticoagulant.

Adolescent↗

Serum dilution neutralization test for California group virus identification and serology.

The serum dilution neutralization test was evaluated for serological diagnosis of California group arbovirus infections and identification of virus isolates. The technical advantages and the degree of subtype specificity of the serum dilution neutralization test over the hemagglutination inhibition test and the complement fixation test were demonstrated with paired specimens from human cases, single human survey sera, and sentinel rabbit sera. Twenty-one virus isolates from various geographical areas of the United States were also used to evaluate the efficacy of the serum dilution neutralization test for specific virus identification.

Adolescent↗

Sensitive neutralization test for virus antibody. 1. Mumps antibody.

A sensitive mumps virus plaque neutralization test has been developed based on the potentiation of virus-antibody complexes by heterologous anti-immunoglobulins (AIG). The enhanced neutralization test was approximately 100 times more sensitive than the conventional neutralization test or the hemagglutination-inhibition test. Using AIG against human immunoglobulin G (IgG) or human IgM permitted determination of the relative titers of the two classes of mumps antibody. The test does not require special equipment or expertise and can be readily introduced in virological laboratories.

Antibodies, Anti-Idiotypic↗