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PubMed1991

[The application of immunofluorescence and neutralization McAb tests to typing diagnosis of 286 polio strains].

The serum neutralization test is a classical method used for typing poliovirus. Since 1987, some type-specific non-neutralization McAb's have been obtained in our laboratory, and were used in indirect immunofluorescence straining tests to detect serotypes I, II and III. A total of 286 poliovirus strains were typed by the indirect immunofluorescence test (IF) and neutralization test (NT). The results demonstrated that both tests were in agreement, suggesting that McAb IF could replace NT as an alternative typing diagnostic method: moreover, IF was easier to perform than NT. It is economical and rapid (the results can be read within 1-2 days as compared to 7 days for NT). The reagent is very stable and can be stored for two years at 4 degrees C or for two months at 37 degrees C.

Antibodies, Monoclonal↗
PubMed1971

Antigenic relationships of equine herpesvirus strains demonstrated by the plaque reduction and neutralization kinetics tests.

The antigenic relationships among 50 strains of equine herpesvirus (EHV) were studied by neutralization tests using antisera prepared in rabbits against four EHV reference strains: types 2 and 3, cytomegalo-like virus 82-A, and our leukocyte isolant H-40. No distinctive antigenic differences among reference strains were demonstrated in reciprocal neutralization tests but each antiserum neutralized its homologous virus more rapidly than any heterologous strain. Forty-six EHV strains isolated from peripheral blood leukocytes of apparently healthy horses were antigenically indistinguishable from each other and from the four reference strains. Their high degree of antigenic relatedness suggests that these viruses are isolants of a single, widely distributed serotype of which type 2 (LK) strain is a typical representative.

Animals↗
PubMed2001

[Epidemiological effects of live attenuated hepatitis A vaccine (H(2)-strain): results of A 10-year observation].

OBJECTIVE: To assess the long-term epidemiological effects after a mass vaccination program, using live attenuated hepatitis A vaccine (H(2)-strain). METHODS: Shengsi county and Jiaojiang city in Zhejiang province were chosen where high incidence rate of hepatitis A was observed. Mass vaccination program of H(2)-strain vaccine was carried out on 1 - 15 year olds. Cohort studies included two groups were carried out in Shengsi. Cases reported program on hepatitis A was under the national standard on clinical diagnosis and partly through laboratory monitor of anti-HAV IgM and IgG, ELISA and Abbott EIA. Population size was estimated by the yearly reports of census bureau of the county and the city. Serum tests including neutralization test, immune-recall responses, and anti-HAV IgG were performed 10 years after the initial immunization. RESULTS: The morbidity of hepatitis A was significantly different between vaccinated groups and non-vaccinated groups (P < 0.01) in the cohort study. The protective efficacy in the site was 100 percent. There was a significantly negative correlation of regression between vaccine coverage and hepatitis A morbidity. In both sites, hepatitis A incidence rate showed decrease of 94.5% and 90.3% in whole population, with 96.9% and 97.9% in 1 - 15 year olds respectively. Neutralization test showed that 63.8% of the serum negative to ELISA after vaccination could still neutralize certain HAV. Immune-recall response after the booster was seen in vaccine group with 80.2% positive rate of anti HAV-IgG, 10 years after immunization among vaccinees in both sites. Not a single hepatitis A case was identified. CONCLUSION: Live attenuated hepatitis A vaccine seemed to have obviously long-term epidemiological effects.

Adolescent↗
PubMedDate not supplied

Comparison of cytotoxicity of various surfactants tested on normal human fibroblast cultures using the neutral red test, MTT assay and LDH release.

We used the neutral red test, MTT assay and lactate dehydrogenase (LDH) release to compare the potential cytotoxicity of six surfactants belonging to different classes--three non-ionic surfactants (Triton x100, octylphenoxypolyethoxy alcohol, from Orion; Tween 60, polyoxyethylene (20) sorbitan monostearate, from ICI Speciality Chemicals; Tween 80, polyoxyethylene (20) sorbitan monolaurate, from Labosi), two anionic surfactants (Texapon K1298, sodium lauryl sulphate, from Henkel; Texapon N40, sodium laurylether sulphate, from Henkel) and one cationic surfactant (benzethonium chloride, from Siber Hegner)--on human fibroblast cultures. According to the LC50 (microg ml(-1)), the tested surfactants can be classified in the following order of increasing cytotoxicity: Tween 80 < Texapon N40 < Tween 60 < Texapon K1298 < Triton x100 < benzethonium chloride.

Benzethonium↗
PubMed2003

Antibodies to ruminant alpha-herpesviruses and pestiviruses in Norwegian cervids.

A serologic survey revealed that Norwegian populations of free-ranging reindeer (Rangifer tarandus tarandus), roe deer (Capreolus capreolus), red deer (Cervus elaphus), and moose (Alces alces) have been exposed to alpha-herpesviruses and pestiviruses. A total of 3,796 serum samples collected during the period 1993-2000 were tested in a neutralization test for antibodies against bovine herpesvirus 1 (BHV-1) or cervid herpesvirus 2 (CerHV-2), and 3,897 samples were tested by a neutralization test and/or enzyme-linked immunosorbent assay for antibodies against bovine viral diarrhea virus (BVDV). Antibodies against alpha-herpesvirus were found in 28.5% of reindeer, 3.0% of roe deer, and 0.5% of red deer, while all moose samples were negative. In reindeer, the prevalence of seropositive animals increased with age and was higher in males than females. Antibodies against BVDV were detected in 12.3% of roe deer, 4.2% of reindeer, 2.0% of moose and 1.1% of red deer. The results indicate that both alpha-herpesvirus and pestivirus are endemic in reindeer and pestivirus is endemic in roe deer in Norway. The viruses may be specific cervid strains. Seropositive red deer and moose may have become exposed as a result of contact with other ruminant species.

Age Factors↗
PubMed2006

Study of the serological response after vaccination against tick-borne encephalitis in Sweden.

The antibody response to vaccination against tick-borne encephalitis (TBE) with FSME-Immun Inject (Immuno AG/Baxter) was studied in 535 persons, mainly adults, attending a vaccination centre in Stockholm, Sweden. Emphasis was laid on long-term follow-up. Antibody activity was measured by three different serological test systems: a commercial ELISA kit, a hemagglutination inhibition (HI) test and a neutralization test (RFFIT). The neutralization test proved to be the most sensitive assay for the detection of the vaccine response, which was demonstrable in the majority of vaccinees (>90% after three and >98% after four and five vaccinations, respectively). ELISA and HI were less sensitive for antibody measurement during primary immunization. Neutralizing antibody activity persisted prior to the third dose in 77% of the vaccinees and prior to the fourth to sixth doses in 89-95% of the vaccinees. ELISA activity, but no neutralizing activity, was found in some individuals. Based on our data and previous experience of vaccine failures after two doses, a more condensed three-dose vaccination schedule may be advantageous and ought to be tested. The persistence of neutralizing antibodies justifies further studies of the antibody responses after the fourth dose for periods beyond the recommended 3-year booster intervals.

Adolescent↗
PubMed2004

Serologic evidence of West Nile virus and St. Louis encephalitis virus infections in white-tailed deer (Odocoileus virginianus) from New Jersey, 2001.

Serum samples from 689 hunter-killed white-tailed deer (Odocoileus virginianus) collected during the 2001 fall hunting season in New Jersey were tested for neutralizing antibodies to West Nile virus (WNV) and St. Louis encephalitis virus (SLEV) by plaque-reduction neutralization tests. WNV-neutralizing antibodies were detected in six (0.9%) of the samples, and SLEV-neutralizing antibodies were found in 11 (1.6%) of the samples. We provide the first report of WNV infection in white-tailed deer.

Animals↗
PubMed1996

Antibodies to arthropod-borne encephalitis viruses in small mammals from southern Florida.

From 1987 through 1991, blood samples were collected from 10 species of small mammals in Indian River Country, Florida (USA). Sera from 1,347 animals were analyzed for hemagglutination-inhibition (HI) antibody to St. Louis encephalitis (SLE) and eastern equine encephalitis (EEE) viruses. Of these, 75 (5.6%) were positive for HI antibody to SLE virus and 121 (9.0%) were positive for EEE antibody. Sera from five mammalian species were tested for neutralizing (NT) antibody to SLE, EEE, Highlands J (HJ a member of the western equine encephalitis virus complex), or Everglades (EVE, a member of the Venezuelan equine encephalitis complex) viruses. By serum neutralization tests, 26 (46%) of 57 had SLE antibodies, 14 (24%) of 58 had EEE antibodies, two (3.2%) of 63 had HJ antibodies, and 9 (14%) of 63 had EVE antibodies. One Sigmodon hispidus and one Peromyscus gossypinus had NT antibodies both to EEE and HJ viruses. Blood samples from 512 mammals were tested for virus. Isolations of one EVE virus and two unidentified arenaviruses were made from P. gossypinus and one EVE virus isolate was made from a S. hispidus.

Animals↗
PubMed2000

The modification of fluorescent antibody virus neutralization (FAVN) test for the detection of antibodies to rabies virus.

The fluorescent antibody virus neutralization test (FAVN) for the detection of antibodies against rabies virus was modified by using a monoclonal anti-rabies antibodies and peroxidase anti-mouse conjugate instead of a fluorescent anti-rabies conjugate. The results were read on an automatic multi-channel spectrophotometer. A total of 182 serum samples from dogs were tested by both the original and modified FAVN methods and the results were compared. Good correlation was found between the two tests. Practically, the modified FAVN test was quicker and could be used for a larger number of samples.

Animals↗
PubMed1977

Cross-neutralization reactions of Escherichia coli and Vibrio cholerae enterotoxins as studied by rabbit skin inoculation test.

Neutralization of enterotoxins of Vibrio cholerae 569 B and Escherichia coli 10407 by antitoxins to V. cholerae 569 B, E. coli 334, 408-3 and 10407 was studies by intradermal inoculation test in the rabbit. Neutralization of V. cholerae enterotoxin by homologous as well as heterologous antisera of E. coli was observed, except that there was no neutralization of the enterotoxin by antiserum to E. coli 408-3 enterotoxin. Neutralization of E. coli enterotoxin to a varied extent by homologous as well as all heterologous antisera, including that of V. cholerae 569 B antitoxin, was also observed.

Animals↗
PubMed1969

Problems of influenza virus vaccine standardization.

The lack of reliable laboratory methods of determining the antigenicity of inactivated influenza virus vaccines prompted a reinvestigation of the reproducibility of the tests used for measuring the antigenic content of influenza vaccines, namely, the CCA and mouse potency tests.The data obtained in the second part of the mouse potency test, i.e., the neutralization test performed either in mice or in embryonated eggs, statistically demonstrated protective differences between 2 vaccines differing in antigenic mass by as little as 2-fold. However, the dependence upon a single egg or mouse neutralization test to provide the correct vaccine/reference ratio assumed more than "biological" variation would allow. Further, the test was long and tedious and it would be impracticable to perform the number of tests needed to obtain statistically significant results. Thus, the extreme variability observed between individual mouse potency tests and the impracticability of performing this test in statistically sufficient numbers precluded its use for measuring antigenic content of inactivated influenza vaccines.The simpler CCA test, on the other hand, did provide the reproducibility required for the correct determination of the vaccine/reference ratio once a stable CCA reference vaccine was prepared. This test was easily reproducible and results obtained were sufficient to allow a meaningful and reliable conclusion to be drawn with respect to vaccine potency.The problems of measuring the relative content of several components in multivalent vaccine preparations and of finding a test which positively correlates with vaccine potency in man, however, remain unsolved.

Agglutination Tests↗
PubMed1980

Rubella immunity testing by mixed hemadsorption.

A mixed hemadsorption (MH) test for naturally acquired rubella immunity and immune responses after rubella vaccination has been compared with the hemagglutination-inhibition (HI) test, the immunodiffusion (ID) test, the neutralization test (NT), the hemolysis-in-gel (HIG) test and the indirect immunofluorescence (IF) test. The MH test was approximately equivalent to the HI test and the NT as to sensitivity for the early response to rubella vaccination, and all three tests are thought to measure protective antibodies, i.e. antibodies supposedly directed against envelope antigens, to about the same extent. The MH test for rubella antibodies is especially suited for laboratories in which this type of test is routinely used for other purposes, e.g. rabies, respiratory syncytial virus, herpes simplex, varicella and certain auto-antibodies. Where the method is already standard it may be preferable to the NT which, although of similar clinical value requires tedious standardization. Both tests may be necessary in cases where the HI test is equivocal or hampered by non-removable on-specific inhibitors.

Antibody Formation↗
PubMed1991

Characterization of monoclonal antibodies against feline infectious peritonitis virus type II and antigenic relationship between feline, porcine, and canine coronaviruses.

Seven monoclonal antibodies (MAbs) with neutralizing activity against feline infectious peritonitis virus (FIPV) strain 79-1149 (type II) were prepared. When the polypeptide specificity recognized by these monoclonal antibodies (MAbs) was investigated by Western immunoblotting, all of the MAbs reacted with peplomer glycoprotein (S) of the virus. By competitive binding assay these MAbs were found to recognize at least 3 different epitopes. The reactivity of these MAbs with 6 viruses classified as FIPV type I (UCD-1, UCD-2, UCD-3, UCD-4, NW-1, and Black), feline enteric coronavirus (FECV) type II strain 79-1683, canine coronavirus (CCV) strain 1-71, and transmissible gastroenteritis virus (TGEV) strains TO-163 and SH was examined by neutralization tests. All MAbs neutralized FECV strain 79-1683, CCV strain 1-71, and TGEV strains TO-163 and SH, while they did not neutralize the 6 FIPV type I viruses. Moreover, the MAb against TGEV strain TO-163, which has strong neutralizing activity against 7 TGEV viruses, neutralized CCV strain 1-71, FECV strain 79-1683, and FIPv strain 79-1146, but did not neutralize the 6 FIPV type I viruses. These results demonstrated that there are at least 3 epitopes involved in the neutralization of FIPV type II strain 79-1146, and that these epitopes are not present in FIPV type I viruses but are present in FECV strain 79-1683 which does not induce feline infectious peritonitis, TGEV strains TO-163 and SH, and CCV strain 1-71. These results suggest the presence of 2 serotypes of FIPV which can be clearly distinguished by the neutralization test using MAbs.

Animals↗
PubMed2004

A test of neutrality and constant population size based on the mismatch distribution.

Several factors including demographic changes, selection, and recombination are known to affect the distribution of the number of pairwise differences between DNA sequences. The effects of each of these forces have previously been used to estimate population parameter values using various assumptions about other factors. In this article, we use the predictions of the mismatch distribution under a standard neutral equilibrium model to design a coalescent simulation-based test and detect any deviation from this equilibrium. When reliable independent estimates are available for the intragenic recombination rate, this test can be used as a neutrality test or a population expansion test in actual studies, under reasonable assumptions.

Animals↗
PubMed1981

Seroepidemiologic studies of acute hemorrhagic conjunctivitis virus (enterovirus type 70) in West Africa. III. Studies with animal sera from Ghana and Senegal.

One hundred and thirty-nine bovine sera collected in Senegal in 1968 and 1969, before the human pandemic of acute hemorrhagic conjunctivitis (AHC), and 145 sera collected in 1977, seven years after the introduction of AHC, were tested for virus neutralizing (VN) titers against enterovirus 70 (EV70) in neutralization tests. Positive rates of VN titers (1:16) were fairly constant (about 40%) in 1968 and 1969 but the proportion of positives from the 1977 collection was significantly higher (61%). The proportion of Ghanaian bovine sera positive in 1977 (39%) was comparable with those collected earlier in Senegal but the proportion was lower in sera of calves (7%). Similar studies were performed on sheep sera collected in 1966, 1967, 1968, and 1969 in Senegal. The proportions positive and the geometric mean titers (GMTs) peaked in alternate years: the proportions were 43% and 54% in 1966 and 1968 but dropped to 12% and 5% in 1967 and 1969, respectively. The prevalence rate for Ghanaian sheep sera in 1977 was comparable to the earlier figures in Senegal. The prevalence of VN (83%) and the GMT (1:27.28) in swine sera in Ghana in 1977 were much higher than those seen in chickens and dogs. However, 87 sera of wild monkeys caught in Senegal after 1970 were negative in neutralization tests. The VN substance detected in these domestic animals was found to be 2-mercaptoethanol sensitive; hence it is considered to belong to IgM. The results seem to favor the hypothesis that enterovirus type 70 (EV70) evolved from an animal enterovirus which shares a common antigen with EV70.

Animals↗
PubMed1974

Antibody to viruses affecting cattle in commercial tissue culture grade fetal calf serum.

Commercial fetal calf serum (FCS) for tissue culture use was tested for neutralizing activity against several viruses which affect cattle. Certain lots of FCS contained no neutralizing activity, whereas other lots contained neutralizing activity to several viruses. It was concluded that the neutralizing activity found in certain lots of sera was due to specific antibody and that its presence could be most easily explained by the contamination of the FCS with serum from postcolostral bovine serum. A nonantibody inhibitor to vesicular stomatitis virus was also found at low levels in most lots of serum. Because those sera which had antibody had antibody to several viruses, it was suggested that the use of the micro-serum neutralization test with a few bovine viruses which are widespread in the bovine population should be satisfactory to detect FCS which was contaminated with postcolostral bovine serum.

Adenoviridae↗
PubMed1986

Purification and partial characterization of a Vibrio hollisae hemolysin that relates to the thermostable direct hemolysin of Vibrio parahaemolyticus.

Hemolysin produced by Vibrio hollisae (Vh-rTDH), which is related to the thermostable direct hemolysin (Vp-TDH) of Vibrio parahaemolyticus, was studied. Vh-rTDH was purified by successive column chromatographies on diethylaminoethyl-cellulose and an immunoaffinity column coupled with anti Vp-TDH immunoglobulin. The purified toxin was homogeneous, as demonstrated by conventional and sodium dodecyl sulfate--polyacrylamide gel electrophoresis (PAGE). The molecular weight of Vh-rTDH was slightly smaller than that of Vp-TDH, as determined by sodium dodecyl sulfate--slab gel electrophoresis. Conventional PAGE also showed a difference between Vh-rTDH and Vp-TDH. Vp-TDH and Vh-rTDH showed different lytic activities on erythrocytes from various animals, in particular chicken, sheep, and calf. The hemolytic activity of Vh-rTDH was heat labile when heated at 70 degrees C for 10 min, unlike Vp-TDH. Immunological cross-reactivity between Vh-rTDH and Vp-TDH was demonstrated by both the Ouchterlony test and neutralization test.

Animals↗