Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Neutralization Tests”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 541 records · Page 30Linked to original sources

Buffalo in the northern Natal game parks show no serological evidence of infection with foot-and-mouth disease virus.

A total of 594 sera collected from buffalo (Syncerus caffer) in the Hluhluwe/Umfolozi Game Reserve complex, Ndumu Game Reserve and the eastern shores of Lake St Lucia were examined for antibody to SAT 1, 2 and 3 types of foot-and-mouth disease (FMD) virus in neutralization tests. No neutralization of SAT 2 or 3 viruses was exhibited by any of the sera tested at final dilutions greater than 10. A small proportion (2,9%) of sera neutralized SAT 1 virus at dilutions up to 10, but these were considered to be due to non-specific reactions. This, together with the absence of clinical FMD in both cattle and game in this region over at least a 45-year period and the failure to isolate FMD virus from pharyngeal scrapings of buffalo sampled in the area, leads to the conclusion that FMD does not occur in these buffalo populations.

Animals↗

Studies on bovine virus diarrhea: serum neutralization, complement-fixation and immunofluorescence.

The complement-fixation, the serum neutralization tests and the fluorescent-antibody technique were the serological methods applied in this laboratory for the detection of antigens for bovine virus diarrhea (BVD). As observed previously, the modified direct complement-fixation (MDCF) test was required to demonstrate antibodies against virus infections of cattle. At a certain stage of infection, the MDCF test was found to be as accurate and less time-consuming than the serum neutralization test for the detection of antibodies in bovine sera. The modified direct complement-fixing antibodies were detectable in the serum from approximately three weeks up to a few months after infection as compared to several years for the serum neutralization test. Thus, as in most other viral diseases, the MDCF test was of value for detecting recent infections while the serum neutralization test detects both recent and long-standing infections. The fluorescent antibody technique was of value to detect viral antigens of both cytopathogenic and noncytopathogenic strains of BVD in primary fetal kidney cell cultures inoculated with field specimens. In addition, the virus was detected in six of 220 fetuses collected at a local slaughter house for the preparation of primary cell cultures. The length of time required for the detection and identification of specific viral antigens by immunofluorescence was considerably reduced over that of the serum neutralization and virus interference tests.

Animals↗

Serological investigations of red foxes (Vulpes vulpes L.) for determination of the spread of tick-borne encephalitis in Northrhine-Westphalia.

Serum samples from 786 red foxes shot between January 1995 and August 1996 in the southern half of Northrhine-Westphalia, located in western Germany, were tested for the presence of antibodies against tick-borne encephalitis (TBE) virus using the Immunozym FSME IgG All Species-ELISA (Immuno, Heidelberg, Germany) as a screening test: 759 sera were negative, 23 (2.9%) were borderline, and four (0.5%) were positive. Nine of the 27 ELISA reactive sera were confirmed by the TBE Western-Blot (Immuno, Heidelberg, Germany). Furthermore these 27 sera were tested for neutralizing antibodies by means of a plaque reduction neutralization test (PRNT) against TBE and West Nile viruses. Only one single serum was found to have a neutralization titre (+1:800 PRNT80) against TBE virus. All other 26 sera were negative for neutralizing antibodies against TBE or West Nile virus. Since the titre of the single serum is low, it can be interpreted that if TBE virus is present, its prevalence is extremely low. Northrhine-Westphalia is not classified as a TBE-endemic area. Further calculated serological testing of game and virological investigation of collected ticks in the affected area seem to be meaningful and necessary.

Animals↗

Examination of the envelope glycoprotein of yellow fever vaccine viruses with monoclonal antibodies.

Two panels of envelope glycoprotein reactive monoclonal antibodies (mAbs) were prepared against yellow fever (YF) 17D vaccine viruses. Five mAbs were prepared against the World Health Organization 17D-204 avian leukosis virus-free secondary seed virus and eight mAbs against 17DD vaccine manufactured in Brazil. The majority of these mAbs were type-specific and displayed differing reactions in neutralization tests. One, B14, would only neutralize YF vaccine virus grown in invertebrate cells. Others would differentiate 17D-204 and 17DD vaccines, from different manufacturers, in neutralization tests when the viruses were grown in vertebrate cells. The data indicate that heterogeneity exists between the epitopes that elicit neutralizing antibody on YF vaccine from different manufacturers.

Animals↗

[Use of erythrocyte diagnostic agents in the universal system of serological reactions].

A method of parallel serologic studies of the material for the detection of specific antigen or antibodies is proposed. This method is not inferior to the known systems of serologic tests (passive hemagglutination-antigen neutralization tests, passive hemagglutination--antibody neutralization tests, etc) in sensitivity and economy, but exceeds all of them in informative value. The method has been tested both experimentally and in the epizootic study of plague infection used as a model. A serologic tests with the use of erythrocyte diagnosticums are based on a universal principle, this method may be used for other infections.

Animals↗

Detection of human immunoglobulins G and M antibodies to Rift Valley fever virus by enzyme-linked immunosorbent assay.

Rift Valley fever virus (RVFV) is an important human and animal pathogen in Africa and has been responsible for infections in travelers. Because of the aerosol infectivity and risk of dissemination of the virus, a need exists for simple, safe, serological tests for diagnosis. An enzyme-linked immunosorbent assay (ELISA) was developed to detect RVFV-specific immunoglobulins (immunoglobulin G [IgG] and IgM). In the test, a betapropiolactone-inactivated, sucrose-acetone-extracted, suckling mouse liver RVFV antigen was captured by mouse RVFV antibodies adsorbed to polystyrene plates. The test sample (human serum) was then added, and the binding of specific antibodies was indicated by alkaline phosphatase-conjugated swine anti-human IgG or IgM. A mu-capture IgM ELISA was also developed by using polystyrene plates coated with goat anti-human IgM incubated successively with serum sample, RVFV antigen, and indicator antibodies. The ELISA for RVFV-specific IgG proved to be more sensitive than hemagglutination inhibition or complement fixation tests and almost as sensitive as the plaque reduction neutralization test in detecting specific antibodies in human sera after vaccination. The two ELISA IgM tests could detect specific IgM antibodies during the first 6 weeks after RVFV vaccination. Three injections of inactivated vaccine were given on days 0, 6 to 8, and 32 to 34. ELISA IgM values for sera obtained on days 6 to 8 were negative or in the lower range of significance, on days 32 to 34 they were strongly positive, and on days 42 to 52 they were waning. Later sera were negative. The plague reduction neutralization test was negative on days 6 to 8 but rose progressively in later samples. These findings suggest that the three doses of RVFV vaccine induce a prolonged primary antibody response. The ELISA IgM could become an important tool for early diagnosis in acute human infection. A number of African sera, some of which were positive for RVFV by plaque reduction neutralization test, were also tested by ELISA IgG. There was good agreement between both tests.

Animals↗

Quantitation of commercial equine tetanus antitoxin by competitive enzyme-linked immunosorbent assay.

In the USA, the potency of commercially prepared equine tetanus antitoxin is determined by the method outlined in the Code of Federal Regulations, Title 9, Part 113.451. In the current test, commercial equine tetanus antitoxin is tested by a toxin neutralization test in guinea pigs. The in vivo test measures antitoxin content through effectiveness of protection of guinea pigs injected with diluted mixtures of antitoxin and a standard toxin. A competitive enzyme-linked immunosorbent assay, designed as an in vitro alternative to the in vivo test, measures antitoxin content based on a competitive reaction between standard or unknown serum and murine monoclonal antibody specific for tetanus toxin. The monoclonal antibody used in the assay delayed death in mouse passive protection studies and reacted with the C fragment of tetanus toxin. No cross-reaction was observed when the antibody was tested with the toxins of Clostridium chauvoei, C. novyi, C. perfringens, or C. sordellii. The in vitro test will measure the antitoxin content of serum samples containing 100-1500 units of antitoxin. Tetanus antitoxin titers obtained by the competitive enzyme-linked immunosorbent assay compared favorably with the toxin neutralization test conducted in guinea pigs. The in vitro assay serves as a feasible alternative to the in vivo test because it can be completed in less time, is reproducible, and eliminates the use of test animals.

Animals↗

Consistent patterns in the contact of domestic animals with tick-borne encephalitis virus in the eastern part of the Russian plain.

Hemagglutination inhibition test (HIT), neutralization test (NT) and complement fixation test (CFT) were used to serologically examine goats and cows in the southern area of the Kirov region and in the Udmurt ASSR. A total of 1641 sera of goats and 1414 sera of cows were examined for the presence on antibodies against tick-borne encephalitis virus. The tests showed a relatively intensive contact of goats and cows with TBE virus in the forests of the eastern part of the Russian Plain: HIT was positive in 27% of goat sera and 17% of cow sera, virus-neutralizing antibodies were detected in 25% of sera of goats and the positivity of CFT was recorded in 32% of goat sera and 29% of cow sera. The relative number of immune animals differed considerably from locality to locality. The localities with great numbers of immune animals were characterized by a distinct increase in the incidence of adult ticks Ixodes persulcatus. The immunization of domestic animals in the natural foci of infection was found to be season-dependent: the highest immunity rates were observed at the end of summer and in the autumn. The seasonal variations in animal immunity are demonstrated in detail on adult goats. Towards the spring time some serum antibodies, namely virus neutralizing antibodies and antihemagglutinins, tended to disappear in a certain number of animals. A similar, but somewhat different pattern of immunization could be observed in cows. In contrast to goats, cows appeared to be less susceptible to the tick-borne encephalitis virus infection, which was substantiated by both serologic data obtain in this study and the data reported in the literature. In the natural foci of TBE virus infection grazing goats and cows had the most intensive contact with the pathogenic agent in the first year of their lives.

Age Factors↗

Immunologic responses to West Nile virus in vaccinated and clinically affected horses.

OBJECTIVE: To compare neutralizing antibody response between horses vaccinated against West Nile virus (WNV) and horses that survived naturally occurring infection. DESIGN: Cross-sectional observational study. ANIMALS: 187 horses vaccinated with a killed WNV vaccine and 37 horses with confirmed clinical WNV infection. PROCEDURE: Serum was collected from vaccinated horses prior to and 4 to 6 weeks after completion of an initial vaccination series (2 doses) and 5 to 7 months later. Serum was collected from affected horses 4 to 6 weeks after laboratory diagnosis of infection and 5 to 7 months after the first sample was obtained. The IgM capture ELISA, plaque reduction neutralization test (PRNT), and microtiter virus neutralization test were used. RESULTS: All affected horses had PRNT titers > or = 1:100 at 4 to 6 weeks after onset of disease, and 90% (18/20) maintained this titer for 5 to 7 months. After the second vaccination, 67% of vaccinated horses had PRNT titers > or = 1:100 and 14% had titers < 1:10. Five to 7 months later, 33% (28/84) of vaccinated horses had PRNT titers > or = 1:100, whereas 29% (24/84) had titers < 1:10. Vaccinated and clinically affected horses' end point titers had decreased by 5 to 7 months after vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: A portion of horses vaccinated against WNV may respond poorly. Vaccination every 6 months may be indicated in certain horses and in areas of high vector activity. Other preventative methods such as mosquito control are warranted to prevent WNV infection in horses.

Animals↗

Preparation and characterization of the monoclonal antibodies against Japanese encephalitis virus.

Six mouse monoclonal antibodies (MoAbs) against Japanese encephalitis virus (JEV) were prepared and analyzed with indirect immunofluorescence assay (IFA), enzyme linked immunosorbent assay (ELISA), haemagglutination inhibition test (HI), neutralization test (NT), antibody dependent cell mediated cytotoxicity (ADCC) assay, antigenic site specific analysis and relative affinity measurement. These MoAbs could be divided into three classes by indirect immunofluorescence cross reactivity among four flaviviruses, 2H4, 2F2, and nG2 were type specific; 2D2 and mC3 were subgroup specific; and mG9 was family specific. 2H4 and 2F2 had higher neutralization activity, 2D2 and mC3 had the function of inducing ADCC effect, mG9 had higher titer in HI. The six MoAbs recognized five antigenic sites on JEV envelope glycoprotein, 2H4 and 2F2 recognized the same or very similar antigenic site and their relative affinity was ranked as: nG2 > 2H4 > 2D2 > mG9 > 2F2 > mC3.

Animals↗

Radiometric cytolysis inhibition assay, a new rapid test for neutralizing antibodies to intact and trypsin-cleaved poliovirus.

We have developed a new rapid test, the radiometric cytolysis inhibition assay (RACINA), for the determination of neutralizing poliovirus antibodies. HeLa cells prelabeled with 51Cr, [3H]leucine, or, preferentially, with [3H]uridine are used as sensitive quantitative indicators of residual infectious virus. Both suspensions and monolayer cultures of the indicator cells can be used. Neutralization of a fraction of a high-titer virus preparation can be scored after the first replication cycle at 8 to 10 h. By lowering the incubation temperature to 30 degrees C, the completion of the cytolysis due to the first replication cycle of poliovirus was delayed beyond 21 h. This makes it possible to use the RACINA, unlike the standard microneutralization assay, for measuring antibodies to trypsin-cleaved polioviruses. The RACINA was found to be as sensitive as and more reproducible than the standard microneutralization assay in the measurement of neutralizing poliovirus antibodies. The RACINA is a rapid and reliable test for neutralizing antibodies and in principle it may be applicable for quantitation of neutralizing antibodies to other cytolytic agents as well.

Animals↗

Characterization of the Palyam serogroup viruses (Reoviridae: Orbivirus).

31 Palyam serogroup viruses were examined by complement-fixation and plaque-reduction neutralization tests and by PAGE of the segmented, double-stranded (ds) RNA genome. Although the viruses were indistinguishable by complement-fixation tests, 10 distinct virus serotypes were identified by plaque-reduction neutralization methods. Palyam group viruses which were distinct by the neutralization test had unique dsRNA profiles, whereas those agents which were indistinct by the neutralization test had identical dsRNA profiles. 20 isolations of 3 Palyam serotypes were made from bovines and Culicoides midges in Australia over a 9-year period. When the genome of these isolates was examined electrophoretically, the dsRNA profiles of virus isolates within a given serotype were identical.

Animals↗

Rapid (24h) neutralization assay for the detection of antibodies to human cytomegalovirus using a monoclonal antibody to an HCMV early nuclear protein.

Neutralizing antibodies to human cytomegalovirus (HCMV) may play an important role during the course of an HCMV infection but yet detection with conventional methods remains difficult and time consuming. A new neutralization test has been developed basically corresponding to the known microtitre technique. A monoclonal antibody to an HCMV early nuclear protein is utilized to detect HCMV (AD169) infected fibroblasts by biotin/streptavidin-enhanced immunoperoxidase staining. The test procedure was thereby confined to only 24 h. Evaluation of the new neutralization test (24h-NT) was done in comparison to the conventional method (NT). Preselected serum samples (n = 217), of which n = 169 were HCMV-seropositive in ELISA (reciprocal titre greater than or equal to 40), were tested for neutralizing antibodies (NAbs) to HCMV. ELISA titres were measured by a commercially available kit (Behring, F.R.G.). NAbs could be detected in 107 sera by 24h-NT (reciprocal titre greater than or equal to 5) and in 110 sera determined by NT (reciprocal titre greater than or equal to 5). Those sera were exclusively positive in ELISA. ELISA negative sera (n = 48; reciprocal titre less than 40) yielded no detectable neutralizing titres. This specific and rapid test should facilitate HCMV-NAbs detection for a wide variety of applications including routine virology diagnostics, evaluation of HCMV hyperimmunoglobulins and medical research.

Antibodies, Monoclonal↗

The 2-deoxy-D-glucose-neutral red test and vagotomy: an experimental study.

The 2-deoxy-D-glucose - neutral red test proved to be successful in assessing completeness of vagotomy in the baboon both intra- and postoperatively. There were no deleterious side-effects and we found that both products could be sterilized adequately. So far we have used this test of 5 patients in the immediate postoperative phase; in 1 patients, on whom the surgeon thought he had done an incomplete parietal cell vagotomy, the test was found to be positive within a week of the operation.

Alkaline Phosphatase↗

Prevalence of bovine viral diarrhea virus infection in bulls in artificial insemination centers in Poland.

This study was directed at the evaluation of the prevalence of bovine viral diarrhea virus (BVDV) infection in bulls in artificial insemination centers. Both serological and virological examinations were performed. Blood samples were tested in micro-seroneutralization test for BVDV antibodies. Virus isolation was performed in cell culture and BVDV antigen was detected in an indirect immunofluorescence assay with monoclonal antibodies. One hundred and seventy-five serum samples and 219 whole blood samples for virus isolation were tested. Neutralizing antibodies were found in 86% of the bulls. Persistent infection (PI) was detected in 0.9% of the analyzed blood samples.

Animals↗

Incubation at raised temperature or enrichment media, combined with secondary enrichment in Rappaport's medium, for the isolation of salmonellas from sewage.

Two FMDV strains which had been previously differentiated by complement-fixation were compared by guinea-pig protection test, kinetic neutralization and micro-neutralization tests. It was found that these tests, which have not been previously applied by the methods described, were all capable of FMDV STRAIN DIFFERENTIATION. Similar differences were found by all methods, which suggests that comparisons made by cross-CF, cross-neutr-alization or cross-protection involve measurement of the same antigen/antibody interactions.

Antibodies, Viral↗

Large envelope glycoprotein and nucleocapsid protein of equine arteritis virus (EAV) induce an immune response in Balb/c mice by DNA vaccination; strategy for developing a DNA-vaccine against EAV-infection.

Equine arteritis virus (EAV) is a member of the Arteriviridae family, that includes lactate dehydrogenase-elevating virus (LDV), porcine reproductive and respiratory syndrome virus (PRRSV), and simian haemorrhagic fever virus (SHFV). Equine arteritis is a contagious disease of horses and is spread via respiratory or reproductive tract. The objective of the present study is to evaluate the possibility for developing a model system for prevention horses against an EAV infection by DNAvaccination. A cDNA bank from the RNA of EAV was established. This gene library contains the translation unit of the EAV open reading frames (ORF) 1 to 7. The identity of the cDNA was confirmed by nucleotide sequence analysis. Using this defined EAV cDNA gene library the cDNA sequence of the viral ORFs were molecularly cloned into the corresponding sites of well characterized and powerful expression vectors (pCR3.1, pDisplay, and/or pcDNA3.1/HisC). The capability of these recombinant plasmids expressing the gene products of the individual viral ORFs 3 to 5, and 7 in induction of an immune response in mouse system was investigated. The Balb/c mice (ten mice per assay) were inoculated with the DNA of the constructed expression vectors harboring and expressing the EAV cDNA of the viral ORFs. The Balb/c mice were injected with about 100 microg DNA diluted in 100 microl PBS. The DNA was injected subcutaneously and into the tibialis cranialis muscle (Musculus gastrocnemius). The mice were boosted 3 to 5 times with the same quantities of DNA and under the same conditions at about two week intervals. Control mice received the same amount of parental expression vectors via an identical route and frequency. The pre- and post-vaccinated sera of the individual animals were screened by neutralization tests (NT). Neutralizing antibodies against EAV were detected when the animals were inoculated with the DNA of the expression vectors harboring cDNA of the EAV ORFs 5 and 7. Highest NT-titers were observed when the animals were administered with the cDNA of ORF 5 and/or with the cDNA of the neutralization determinants of EAV that is located on the N-terminal ectodomain of the gene product of ORF 5 between the amino acid positions 1-121. These results obtained from these studies justified proofing the capability of the EAV cDNA sequences of the viral genes including ORFs 5 and 7 in the autologous animal system horse.

Animals↗