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PubMed1994

Measles virus-specific functional antibody responses and viremia during acute measles.

Antibody titers measured in functional and immunofluorescent assays were compared with proportions of peripheral blood mononuclear cells infected with measles virus in 8 adults with measles. In addition, a syncytium inhibition assay (SIA) for measuring neutralizing antibody using low-passage virus was compared with a standard plaque neutralization test (PNT). Antibody-dependent cellular cytotoxicity (ADCC) antibody rose later but attained higher titer than neutralizing, antibody-dependent complement-mediated lysis, IgM, or IgG antibodies. When titer changes between specimens from each patient obtained on different days were compared, only ADCC (r = .81, P = .026) and IgM (r = .81, P = .027) antibodies correlated with reductions in viremia. SIA and PNT correlated well (r = .93, P < .001). ADCC may be an important defense against measles. The delay in ADCC antibody relative to other antibodies is unique among viruses studied. The SIA is a useful alternative to the PNT for measuring measles neutralizing antibody.

Acute Disease↗
PubMed1991

Characterization of monoclonal antibodies to bovine viral diarrhoea virus: evidence of a neutralizing activity against gp48 in the presence of goat anti-mouse immunoglobulin serum.

Twenty-one monoclonal antibodies (MAbs) directed against the NY-1 and the Osloss-c strains of bovine viral diarrhoea virus were produced and characterized by indirect immunofluorescence assay, radioimmunoprecipitation and neutralization tests. Fourteen MAbs directed against the NY-1 strain recognized the gp48 and showed a weak neutralizing activity in the presence of goat anti-mouse immunoglobulin serum.

Animals↗
PubMed1982

Comparison of enzyme-linked immunosorbent assay and neutralization techniques for measurement of antibody to respiratory syncytial virus: implications for parenteral immunization with live virus vaccine.

The sensitivity of an enzyme-linked immunosorbent assay (ELISA) to detect low levels of antibody to respiratory syncytial (RS) virus was compared with a tube dilution neutralization test (NEUT) on sera obtained from children who received a parenteral live RS virus vaccine. Among the children who developed antibody in response to live RS virus vaccine. ELISA was as sensitive as NEUT at detecting antibody increases. Some children who did not have detectable prevaccine ELISA antibody possessed NEUT antibody; these children were generally less than 12 months old, suggesting that they had low levels of maternal antibody. Low levels of NEUT or ELISA antibody were associated with the absence of antibody increases after injection of live RS virus vaccine. The quantity of antibody stimulated by this live RS virus vaccine was small compared with that which was stimulated by naturally acquired RS virus infection. We concluded that ELISA is a satisfactory test for determining antibody to RS virus in vaccine field trials, given the understanding that low levels of preexisting antibody are not detected in some instances.

Antibodies, Viral↗
PubMed1987

Effect of monoclonal antibody to pertussis toxin on toxin activity.

Two distinct monoclonal antibodies, one to pertussis toxin subunit S2, called 9G8, and another to subunits S2 and S3, called 11E6, were generated from the hybridomas of myeloma SP2/0 and spleen cells of BALB/c mice immunized mainly with the subunit S234 complex. Binding ability of 9G8 and 11E6 to the subunits was confirmed by the enzyme-linked immunosorbent assay and immunoblotting analysis. Generation of 11E6 bound to both S2 and S3 might mean that there is common antigenicity between S2 and S3. Neutralizing activities of 9G8 and 11E6 on various biological activities of pertussis toxin, including ADP-ribosyltransferase and leukocytosis-promoting, islet-activating, permeability-increasing. Chinese hamster ovary (CHO) cell-clustering, and hemagglutinating activities, were compared with those of anti-S1 monoclonal antibodies 1B7 and 3F10, which were isolated and characterized in a previous study (H. Sato, A. Ito, J. Chiba, and Y. Sato, Infect. Immun. 46:422-428, 1984). 1B7 and 3F10 neutralized ADP-ribosyltransferase activity of pertussis toxin or S1, but 9G8 and 11E6 did not. 1B7 showed very potent neutralization against leukocytosis-promoting, islet-activating, permeability-increasing, and CHO cell-clustering activities of pertussis toxin, but 3F10 did not, although anti-ADP-ribosyltransferase activities of both antibodies were identical. 11E6 neutralized leukocytosis-promoting, islet-activating, CHO cell-clustering, and hemagglutinating activities but not permeability-increasing activity. 9G8 showed slight neutralization of leukocytosis-promoting and CHO cell-clustering activities. Specific activities of 1B7 and 11E6 in each neutralization test were higher than or almost comparable to those of polyclonal antibodies to pertussis toxin. The neutralizing mechanism of 1B7 and 11E6 in leukocytosis-promoting activity was compared. 11E6 seemed to interfere with the binding of pertussis toxin to receptors on mouse spleen cells.

Animals↗
PubMed1970

Two African viruses serologically and morphologically related to rabies virus.

Lagos bat virus and an isolate from shrews (IbAn 27377), both from Nigeria, were found to be bullet-shaped and to mature intracytoplasmically in association with a distinct matrix. They were related to, but readily distinguishable from, rabies virus and each other by complement fixation and neutralization tests. The three viruses, including rabies, form a subgrouping within the rhabdoviruses.

Animals↗
PubMed1980

Antigenic relationship between H1 and H2 of influenza A virus.

Using cloned viruses of proven purity, and by the methods of hemagglutination inhibition, single radial hemolysis, strain-specific complement fixation and neutralization tests we have demonstrated the serological cross reaction between late H1N1 variant (Dutch/56) and H2N2 of influenza A virus with fowl and hamster antisera. Such a cross reaction is not detected with earlier H1N1 variants. Serological crossing covers variants of H2N2 virus isolated from 1957-1966 but in decreasing titers, and disappears with the last variant of H2N2 isolated late in 1967. Analysis with mono-specific antisera or antigens prepared with recombinants reveal that the hemagglutinins of late H1N1 and H2N2 are related, while their neuraminidases are distinct. We have discussed the bearing of such antigenic relationships to previous epidemiological observations on the partial protection of patients convalescent from late H1N1 disease against H2N2 and to the recombination theory for the origin of H2N2 virus.

Animals↗
PubMed1993

Mapping of functional epitopes of Japanese encephalitis virus using monoclonal antibodies.

Epitopes involved in the important functions, hemagglutination (HA) and neutralization (NT), were mapped on Japanese encephalitis (JE) virus proteins by using monoclonal antibodies (MAbs). Fourteen MAbs raised against Nakayama-Yoken strain of JE virus characterized by hemagglutination inhibition (HI) and plaque reduction neutralization test (PRNT) were used to map the epitopes on the JE proteins by Western blot analysis in which non-reducing conditions were used for sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). With these MAbs, at least 8 functional epitopes were demonstrated comprising (i) epitopes recognized by 5 MAbs which gave strong HI but weak NT activities and were mapped on the envelope (E) 53 kDa protein; (ii) epitopes recognized by 2 MAbs which showed weak HI but strong NT activities and were mapped also on the E protein; (iii) epitopes recognized by 2 MAbs which possessed weak HI but no NT activities and were mapped on the E protein; (iv) an epitope recognized by 1 MAb which gave weak NT and no HI activities and was mapped on the nonstructural protein 5 (NS5); (v) an epitope recognized by 1 MAb which showed activities similar to (i) but was mapped on both E and NS5; (vi) an epitope recognized by 1 MAb which had high activities to both HI and NT and was mapped on E and NS5; (vii and viii) epitopes recognized by 1 MAb which also gave low HI but high NT, and strong HI as well as strong NT activities respectively, but their location could not be demonstrated by SDS-PAGE under non-reducing condition.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Comparison of techniques for demonstrating antibodies to Rift Valley fever virus.

Nine serological techniques were compared by monitoring the response to infection with Rift Valley fever (RVF) virus in three sheep. Antibodies were monitored daily for the first 14 days after infection, then weekly and later fortnightly up to week 24. The earliest antibody response was detected in one sheep on day 3 by a plaque reduction neutralization test, and by day 6 antibodies were demonstrable in all three sheep by haemagglutination-inhibition, reversed passive haemagglutination-inhibition, immunodiffusion, indirect immunofluorescence (IF), enzyme-linked immunosorbent assay and neutralization of cytopathic effect in cell cultures. Antibodies were demonstrable by complement fixation on day 8 at the earliest. IF and the two neutralization techniques produced the highest titres, but all tests could be used satisfactorily for the serological diagnosis of RVF. Inactivated antigen could be used for all except the neutralization tests. A radioimmunoassay technique using 125I-labelled staphylococcal protein A detected antibodies on day 8 at the earliest and produced lower mean titres than some of the other techniques. This was probably because sheep immunoglobulins bind protein A poorly.

Animals↗
PubMed2003

Modelling the adsorption kinetics of erythromycin onto neutral and anionic resins.

In this study the selective adsorption method was chosen to enable the recovery of erythromycin. The following sorbents were tested: neutral resins (XAD-4, XAD-7 and XAD-16) and an anionic resin (IRA-410). A mathematical kinetic model for the adsorption of erythromycin against time, on XAD-4, XAD-7 and XAD-16 resins, is proposed. Both Freundlich and Langmuir models showed a good fit for the sorbents XAD-7 and IRA-410 resins. The highest adsorption efficiency was observed when synthetic neutral resin, XAD-7 and XAD-16, were used. The estimated affinity and concentration factors show that the neutral resins tested are adequate for the selective adsorption of erythromycin. The estimated values of enthalpy and free energy of adsorption, lower than 12 kJ mol(-1) and -2 kJ mol(-1), respectively, indicate that a physiosorption process occurred.

Acrylic Resins↗
PubMed1995

Antibodies to hemorrhagic fever viruses in domestic livestock in Niger: Rift Valley fever and Crimean-Congo hemorrhagic fever.

A repository of domestic animal sera collected in Niger between 1984 and 1988 was assayed for antibody against two zoonotic hemorrhagic fever viruses known to be present in the West African Sahel. A total of 2,540 serum samples from 2,324 cattle, sheep, goats, and camels were tested by an IgG-specific enzyme-linked immunosorbent assay (ELISA) and the 80% plaque reduction neutralization test (PRNT80) for Rift Valley fever (RVF) virus antibody. Of the 2,540 sera tested for RVF-specific IgG antibody, 1,676 sera from cattle, sheep, and goats were examined for RVF-specific IgM antibody by ELISA. A subset of 2,263 sera were examined for evidence of Crimean-Congo hemorrhagic fever (CCHF) virus antibody by an IgG-specific ELISA. Antibody against CCHF virus was found to be most prevalent in adult cattle (422 of 732 or 57.7% positive) sampled at nine locations in the Niamey area. The highest prevalence for RVF neutralizing antibodies was found in camels from the Agadez Department with 67 (47.5%) of 141 positive. The results indicate that both CCHF and RVF viruses are circulating in Niger and are potential zoonotic health risks.

Animals↗
PubMed2002

Antigenic relationships between six genotypes of the small hydrophobic protein gene of mumps virus.

Six different genotypes of mumps virus, A, C, D, G, H and I, genotyped on the basis of the small hydrophobic protein gene sequence, were subjected to antigenic comparison. Monoclonal antibodies directed against the haemagglutinin-neuraminidase protein of the SBL-1 strain of genotype A were used in immunofluorescence tests with different mumps virus strains. In addition, the six virus genotypes were compared by cross-neutralization tests with human post-vaccination sera after vaccination with the Jeryl Lynn (JL) strain of mumps virus and with rabbit hyperimmune sera directed against the A or D genotypes of mumps virus. Genotypes C, D, G, H and I could not be antigenically separated. In contrast, three different virus strains of genotype A, SBL-1, JL and Kilham, were distinct and were found to represent three different serotypes within the A genotype of mumps virus. Vaccination of Swedish children with the JL strain of mumps virus resulted in clearly lower neutralization titres against the SBL-1 strain, which is endemic in Sweden, compared to the homologous vaccine titres.

Amino Acid Sequence↗
PubMed1999

A simple, rapid and reliable enzyme-linked immunosorbent assay for the detection of bovine virus diarrhoea virus (BVDV) specific antibodies in cattle serum, plasma and bulk milk.

To detect Bovine Virus Diarrhoea Virus (BVDV)-specific antibodies in cattle serum, plasma and bulk milk, a simple, reliable and rapid blocking ELISA ("Ceditest") has been developed using two monoclonal antibodies ("WB112" and "WB103") directed to different highly conserved epitopes on the non-structural peptide NS3 of pestiviruses. The test can be performed at high reproducibility using undiluted samples. In testing 1000 field serum samples, the ELISA showed a specificity and a sensitivity relative to the virus neutralization test of 99% and 98%, respectively. The blocking ELISA is able to detect specific antibodies in serum obtained 12 days after an acute infection and in serum of vaccinated and challenged animals. A frequency distribution diagram, obtained by testing almost 1800 random Dutch field serum samples, showed a clear separation between a negative population (maximum frequency of the % inhibition at -5%) and a positive population (maximum frequency of the % inhibition at 95%). Based on these data, the prevalence of seropositive animals was 65% (95% confidence interval: 63%-67%). By exchanging plasma- and bulk milk samples between two laboratories (The Netherlands and Denmark), a good overall agreement was found between results obtained with the Ceditest and those obtained with the Danish blocking ELISA as used in the Danish BVDV eradication programme. The results indicate that BVDV infections can reliably be diagnosed by the Ceditest ELISA and that the test is suitable for use in large scale screening and eradication programmes.

Animals↗
PubMed1989

Characterization of monoclonal and polyclonal antibodies to bovine enteric coronavirus: establishment of an efficient ELISA for antigen detection in feces.

Monoclonal antibodies to bovine enteric coronavirus (BEC) were produced. Additionally, polyclonal antibodies were made in rabbits and guinea pigs and extracted from the yolk of immunized hens. The antibodies were characterized by neutralization test, hemagglutination inhibition test, enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Neutralizing antibody titers of polyclonal antisera ranged from 1:1280 to 1:40,000. Only one out of 908 hybridoma colonies tested secreted antibodies with neutralizing activity. By ELISA, polyclonal sera exhibited high background reactions that could be significantly reduced by treatment with kaolin in the case of rabbit sera. Attempts to establish an ELISA for BEC antigen detection based on polyclonal sera failed due to low sensitivity and specificity. Optimal results were achieved when a mixture of two monoclonal antibodies was coated onto microplates for antigen capture, while rabbit hyperimmune serum served as detecting antibodies in an indirect assay. The combination of the two monoclonal antibodies did not increase sensitivity synergistically, but in a compensatory fashion, probably because of epitope differences between BEC field strains.

Animals↗
PubMed1976

[Use of indirect complement fixation test for studying foot and mouth disease virus].

The indirect complement-fixation test was used in experiments for the subtype differentiation, and with the agar gel diffusion and the serum neutralization test was studied the immunity in cattle that had already recovered from foot-and-mouth disease or were immunized against the disease. It was found that the indirect complement-fixation test is instrumental in the demonstration of the antigenic differences between the foot-and-mouth viruses. Comparative experiments have shown that the indirect test in this case is more expedient and of higher economic value than the direct CF reactions after Traub and Möhlmann and after Marucci. The assessment of the results of comparative investigations of sera carried out by the indirect complement-fixation test, the agar gel diffusion test, and the serum neutralization reaction revealed that the indirect reaction is a convenient, quick, and prompt method of work. Discussed is the sensitivity of the reaction and its advantages and its advantages in studies on a large scale on the immune status of animals immunized against F.M.D. or animals that have recovered from the disease.

Animals↗
PubMedDate not supplied

Prevalence of antibodies to infectious bursal disease virus serotypes I and II in 75 Ohio chicken flocks.

Serum samples from 75 Ohio layer flocks were tested for antibodies to infectious bursal disease virus (IBDV) serotypes I and II using a virus-neutralization test. Infectious bursal disease had been diagnosed in three of the 75 flocks. Antibody titers to serotype I were detected in 77% of the flocks, and antibody titers to serotype II were detected in 47% of the flocks. Of the eight flocks known to have been vaccinated using a serotype I IBDV vaccine, all had antibody titers to serotype I, and four had antibody titers to serotype II. Eleven of 17 known unvaccinated flocks had antibodies to serotype I, and seven had antibodies to serotype II.

Animals↗
PubMed1976

Evaluation of the hemolysis-in-gel test for the screening of rubella immunity and the demonstration of recent infection.

The hemolysis-in-gel method for detection of antibodies to rubella virus gave results which correlated well with results of hemagglutination inhibition and neutralization tests. With a diffusion time of 24 or 48 h, a linear correlation was obtained between the logarithm of antibody concentration and the diameter of the hemolytic zone. Fourfold, and even twofold, differences in serum antibody concentrations were shown to give statistically significant differences in hemolytic zone diameters. It could therefore be concluded that the hemolysis-in-gel test is well suited for the serological diagnosis of primary rubella infection, as well as of reinfection. The sensitivity of the hemolysis-in-gel test was comparable to that of the hemagglutination inhibition test. Pigeon erythrocytes were superior to sheep erythrocytes for use in the test. Studies of the antibody response after natural rubella infection or vaccination showed that the appearance and persistence of antibodies measured by hemolysis in gel is similar to that of hemagglutination inhibition antibodies.

Animals↗
PubMed1971

A micro-passive hemagglutination test for the rapid detection of antibodies to infectious bovine rhinotracheitis virus.

Serum antibodies to infectious bovine rhinotracheitis virus are most commonly detected using the serum-virus-neutralization test, a time-consuming and expensive procedure. A passive hemagglutination test was developed for the rapid detection of antibodies using the Microtiter System. Two cloned strains of the virus were used in the investigation, namely the BF strain and the Los Angeles strain. Erythrocytes from several species were used. The best results were obtained using the Los Angeles strain of virus adsorbed to sheep cells treated with formalin and tannic acid. Sera from field cases of the disease as well as hyperimmune bovine and rabbit sera were tested using both the micro-passive hemagglutination test and the standard plaque-neutralization procedures. Comparable titers were obtained in the two tests. The micro-passive hemagglutination test is rapid, accurate and economical.

Animals↗