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PubMed1978

Serologic and cross-immunity studies with contagious ecthyma and goat pox virus isolates from the western United States.

Contagious ecthyma and goat pox viruses were isolated from goats affected with papulopustular epidermal lesions. Results of in vitro serum neutralization and in vivo cross-immunity studies indicate that these contagious ecthyma and goat pox isolates from the western United States are antigenically dissimilar and that exposure or vaccination with one isolate could not be used as a method of inducing immunity to the other. Exposure to each isolate conferred immunity to re-exposure with the same agent. The serum neutralization test indicates that the severity and persistance of lesions to goat pox infection influences the nature of the antibody response to an initial exposure.

Animals↗
PubMed1995

A monoclonal antibody against tetrodotoxin that reacts to the active group for the toxicity.

A monoclonal antibody against tetrodotoxin was produced. Tetrodotoxin coupled with keyhole limpet hemocyanin was used as an immunogen to BALB/c mice. These mice had no clinical signs for the toxicity of tetrodotoxin during the immunization. The reason may be that the guanidyl group of tetrodotoxin which is an important group for the toxicity was hidden by coupling with keyhole limpet hemocyanin. The monoclonal antibody was highly specific for tetrodotoxin and had no cross-reaction to tetrodotoxin derivatives, paralytic shellfish toxins, keyhole limpet hemocyanin and crude proteins from various organs of puffer fish. Also, tetrodotoxin was neutralized in vitro by this antibody. From the fact that the structural difference between tetrodotoxin and anhydro-tetrodotoxin is recognized by this antibody, it was suggested that this antibody reacted with the OH-groups on C-4 and/or C-9 of tetrodotoxin. In addition, the results from immunization and neutralization tests demonstrated that tetrodotoxin became non-toxic even when one of the active groups of tetrodotoxin was coupled by a molecule.

Animals↗
PubMed1977

Heterogeneity among strains of Mycoplasmatales virus-laidlawii 2.

Five isolates of Mycoplasmatales virus-laidlawii 2 (MV-L2) derived from bovine strains of Acholeplasma laidlawii were shown to differ in host range, plaque morphology and neutralization tests with MV-L2 antiserum. Cross-testing using virus resistant clones of A. laidlawii confirmed the heterogeneity of this group. Adaptation of viruses to sub-optimal host was demonstrated.

Acholeplasma laidlawii↗
PubMed1998

Use of monoclonal antibodies to identify serotypes of enterovirus isolates.

Nonpoliovirus enteroviruses cause a variety of diseases that are common in young children and adults. The "gold standard" for laboratory diagnosis of enteroviruses is cell culture isolation, followed by serotype identification by neutralization assay. These procedures are time-consuming and expensive. Rapid serotype identification of enteroviruses is important in differentiating nonpoliovirus enterovirus pathogens from vaccine strain polioviruses that can be shed for some time after vaccination. In the present investigation, we evaluated a rapid method for serotype identification of enteroviruses by indirect immunofluorescence assay (IFA) using commercially available monoclonal antibodies for polioviruses, coxsackieviruses type B, and six serotypes of commonly circulating echoviruses. Of 291 isolates of enteroviruses included in the study, 95 were polioviruses and 196 were nonpoliovirus enteroviruses. Two hundred thirty-four of these (38 polioviruses and 196 nonpoliovirus enteroviruses) were consecutively grown in the laboratory over a 5-year period. IFA identified the serotypes of 74% of the consecutive isolates and 71% of all enterovirus isolates by yielding a positive staining result. The levels of agreement in the identification of the enterovirus group between IFA and neutralization tests were 92% for consecutively grown isolates and 85% for all enterovirus isolates. The sensitivity of the IFA for the detection of viruses for which specific monoclonal antibodies were applied was 73% for polioviruses, 85% for coxsackieviruses type B, and 94% for echoviruses. Specificity was near 100% for polioviruses and coxsackieviruses type B and 94% for echoviruses. We conclude that IFA can be helpful as a preliminary test for serotype identification of enteroviruses. The results are most accurate when the test identifies the isolate as a poliovirus.

Antibodies, Monoclonal↗
PubMed1984

Clostridium botulinum subtype Ba.

Strain 657 has been described as a toxin variant of Clostridium botulinum type B. Neutralization tests performed with types A and B botulinal antitoxins of known potency and avidity at 20, 25, 50, 100, 200, 2,000 and 20,000 mouse LD50 levels of testing, have shown that 657 toxin is a mixture of B (approximately 95% of the complex) and A antigenic fractions. The possibility of a cross-contamination between A and B serotypes has been practically ruled out through the serologic screening of the toxins from 100 well isolated colonies taken from two colony variants of this strain. Strain 657 rabbit antitoxin possesses A and B neutralizing activities. Strain B 657 produces a hitherto undescribed complex toxin and it represents the prototype of a new C. botulinum serotype, subtype Ba.

Animals↗
PubMed1984

Poliomyelitis outbreak in South Africa, 1982. II. Laboratory and vaccine aspects.

Between May and September 1982 an extensive epidemic of poliomyelitis occurred in Gazankulu in the north-eastern region of South Africa, with a total of 260 paralysed cases and 42 deaths. A three-part study was instituted, the first of which established the aetiological agent of the epidemic to be poliovirus type 1. This was determined initially by serology and later by isolation of the virus, chiefly from stool specimens. The potency of the trivalent oral polio vaccine manufactured at the National Institute for Virology, was evaluated in the second study by in vitro potency testing of samples recalled from the epidemic areas and also in vivo by determining the serological response of seronegative children immunized with fresh vaccine. Nearly half of the vaccine samples withdrawn from the field had sub-optimal titres whereas fresh vaccine produced a very satisfactory serological response. In order to plan immunization rationally, the third study, namely the determination of the immune status of the various populations throughout the country was instituted. This involved both serology, using a neutralization test on randomly collected sera and also an analysis of patient history and health card documentation. With the exception of the urban black area (Soweto), which had high levels of immunity both serologically (75%) and on history, other areas examined had poorer levels of immunity of approximately 52% on serological testing and 57% on history. It is clear that the epidemic was multifactorial in origin, with fall-offs in the level of immunization and evidence of breaks in the cold chain being the major factors.

Antibodies, Viral↗
PubMed1990

Molecular mass, biochemical composition, and physicochemical behavior of the infectious form of the scrapie precursor protein monomer.

A highly purified fraction obtained from scrapie (263-K strain)-infected hamsters' brains by an alternative procedure without proteinase K treatment contained a protease-resistant form of the scrapie precursor protein (PrPSc) and infectivity of 9.9 +/- 0.7 log LD50/ml. Polyclonal antibodies produced against hamster scrapie amyloid protein (PrP27-30) and used in a neutralization test diminished infectivity of the PrPSc preparations by 1.6 log after intracerebral inoculation and by 1 log after intraperitoneal inoculation. PrPSc was subjected to size-exclusion HPLC; greater than or equal to 60% of the eluted infectious units were recovered from the peak with an apparent mass of 30.4 +/- 0.6 kDa. Characterization by UV absorption spectra, SDS/PAGE, immunoblots, N-terminal amino acid sequence, and neutral sugar and amino sugar analyses demonstrated homogeneity of the infectious units. The neutral sugar and amino sugar compositional analyses revealed high mannose, glucosamine, fucose, and sialic acid content. This demonstrated an extensive posttranslational modification by the complex type of N-linked glycosylation and glycane core of C-terminal glycolipid of PrPSc. The results correspond to the predicted size, composition, and sequence of PrPSc and indicate that this protein may be the only component of scrapie infectious unit or the infectious form of scrapie precursor.

Amino Acid Sequence↗
PubMed1976

Antigenic relationships of alphaviruses by a simple micro-culture cross-neutralization method.

A cross-neutralization study of 22 alphaviruses disclosed two major antigenic complexes and two viruses that were distinct from all the others. Cross-reactions were common but were restricted within the complexes. Sub-groupings within a complex were also shown, and some viruses proved to be indistinguishable by neutralization testing. The results generally paralleled previously reported data. Investigations were carried out, using BHK 2I cells in a micro tissue-culture system, on all available alphaviruses.

Antigens, Viral↗
PubMed2001

Genomic sequencing of deer tick virus and phylogeny of powassan-related viruses of North America.

Powassan (POW) virus is responsible for central nervous system infection in humans in North America and the eastern parts of Russia. Recently, a new flavivirus, deer tick (DT) virus, related to POW virus was isolated in the United States, but neither its pathogenic potential in human nor the taxonomic relationship with POW virus has been elucidated. In this study, we obtained the near-full-length genomic sequence of the DT virus and complete sequences of 3 genomic regions of 15 strains of POW-related virus strains. The phylogeny revealed 2 lineages, one of which had the prototype POW virus and the other DT virus. Both lineages can cause central nervous system infection in humans. By use of the combination of molecular definition of virus species within the genus Flavivirus and serological distinction in a 2-way cross-neutralization test, the lineage of DT virus is classified as a distinct genotype of POW virus.

3' Untranslated Regions↗
PubMed1978

Serodiagnosis of herpesvirus infection in primates.

The presence of herpesviruses in various animal tissues used for vaccine production is of importance, inasmuch as these viruses may go unrecognized, residing in latent form. Of greatest concern is Herpesvirus simiae (B virus) present in the majority of macaca species, one of the more extensively used laboratory nonhuman primates. The close antigenic relationship of B virus to human herpes strains (H. hominis) and SA8 (a baboon herpesvirus) makes serologic differentiation extremely difficult, but necessary. Complicating the differential diagnosis is the frequent infection of simian species with human strains. The present study clearly shows that many animal sera negative to herpes simplex will be negative to B virus in the neutralization test. However, a sufficient number of sera will be negative to simplex, but positive to B virus, indicating that the assumed one-way cross is invalid. Also, if antibody to simplex is present, and this antibody is present in most adult macaques, then differentiation requires testing with B virus. At least 50% of these positive sera may appear to be negative for B virus if only tested with H. hominis. Approximately 25% of the remaining positive sera will have higher titers to B virus than to herpes simplex and 25% will have the same titers to both viruses. Further, complement is required by many of the test sera to detect the presence of antibodies to B virus.

Animals↗
PubMed1995

The glycoprotein of viral hemorrhagic septicemia virus (VHSV): antigenicity and role in virulence.

In order to study the antigenic structure of the G protein of VHSV, we produced several anti-G monoclonal antibodies (MAbs) and used 4 neutralizing MAbs (NMAbs) to select resistant (MAR) mutants. Each MAR mutant was confronted with the 4 NMAbs in a neutralization test, and also with our panel of MAbs in surface plasmon resonance (SPR) analysis to determine the extent of their relatedness. Determination of the sequence of the entire G gene of representative MAR mutants allowed us to map the mutations responsible for the resistant phenotypes. We identified several locations on the G protein sequence, which represent, most probably, critical positions within the binding sites of the neutralizing MAbs. In addition, the MAR mutants selected with a cross-reactive MAb exhibited a reduced pathogenicity for fish. This indicated that the regions bearing the point mutations selected with MAb C10 were probably involved in the determination of the virulent phenotype.

Amino Acid Sequence↗
PubMed1994

Serologic and virologic evidence of a Prospect Hill-like hantavirus in Wisconsin and Minnesota.

The purposes of this study were to determine if hantaviruses were present in the Great Lakes port areas of Wisconsin and Minnesota and if so, to identify which virus and which rodent host species were involved. Rodents were trapped in Duluth, Minnesota, Superior, Green Bay, and Milwaukee, Wisconsin, all ports of call for international maritime shipping. A total of 675 wild rodents were captured and tested, including 310 meadow voles (Microtus pennsylvanicus), 173 Norway rats (Rattus norvegicus), 179 Peromyscus spp., (including white footed mice [P. leucopus] and deer mice [P. maniculatus gracilis and P. maniculatus bairdii]), and 13 house mice (Mus musculus). Twenty percent of the rats, 17% of the meadow voles, 8% of the house mice, and 3% of the Peromyscus spp. had antibody to a hantavirus by immunofluorescent antibody assay (IFA). By the plaque-reduction neutralization test (PRNT), nine of 36 meadow voles, one of 4 P. leucopus, and one of 34 rats had hantavirus antibody, with the highest titers to Prospect Hill (PH) virus. All of the PRNT-seropositive individuals were from the twin cities of Superior and Duluth. Hantavirus antigen was detected in lung tissue by IFA in M. pennsylvanicus and Peromyscus spp., but not in rats. Two hantaviruses, designated SD-1 and SD-2, were isolated from M. pennsylvanicus captured in Duluth and found to be very similar to prototype PH virus by cross-IFA and cross-PRNT. Virus isolation attempts were unsuccessful from tissues of the Peromyscus spp. and the rats.

Animals↗
PubMed1987

Evaluation of an enzyme-linked immunosorbent assay for the detection of transmissible gastroenteritis virus antibodies.

An enzyme-linked immunosorbent assay (ELISA) using a detergent-solubilized antigen of purified virus was developed for detection of antibody against porcine transmissible gastroenteritis (TGE) virus in swine serum. The ELISA demonstrated antibody responses in pigs immunized intramuscularly with the attenuated TO-163 strain of TGE virus and in pigs orally infected with the virulent Shizuoka strain of the virus. The results of the ELISA were well correlated with those of the neutralization test. These results indicate the usefulness of the ELISA as a serological tool for TGE virus antibody.

Animals↗
PubMed1976

Comparative studies of some African arboviruses in cell culture and in mice.

Twenty African arboviruses, five alphaviruses, nine flaviviruses, three Bunyamwera Group viruses, two Bwamba Group viruses and one ungrouped virus were titrated in parallel in 11 cell systems in suckling mice and adult mice. The relative sensitivities of the in vitro and in vivo systems have been compared. The highest infectivities were obtained in suckling mice. Vero and LLC-MK2 cells produced plaques with the greatest number of viruses and Semliki Forest virus grew most readily. Ntaya virus and Dengue 1 virus were difficult to culture in vitro and Zika virus yielded better in cell culture than in adult or suckling mice. In vitro and in vivo neutralization tests were made on human sera in groups of 50. Each group of sera was tested against one of five viruses, representative of three of the arbovirus groups titrated. Good agreement was obtained between the two test systems with West Nile, O'nyong-nyong and Wesselsbron viruses but there were significant differences in results obtained with Germiston and Pongola viruses.

Africa↗
PubMed1984

Radial immunodiffusion enzyme assay for detection of antibodies to pseudorabies virus in swine serum.

A radial immunodiffusion enzyme assay for the detection and quantitation of antibodies to pseudorabies virus in swine sera was developed and the methods were standardized. The assay combined the principle of radial immunodiffusion with enzyme-linked immunosorbent assay. Quantitation of pseudorabies virus antibody titers was determined by measuring the diameter of a colored circular zone after overnight incubation of antibody with antigen. The specificity and sensitivity of the radial immunodiffusion enzyme assay were compared with that of the standard virus-neutralization test, and the results were determined to be correlated highly (r = 0.694, P less than 0.0001). The assay also appeared to be highly reproducible and simple to perform.

Animals↗
PubMed1984

Seroepidemiological survey on dengue and Japanese encephalitis virus infections in Asian monkeys.

To investigate the ecology of dengue and Japanese encephalitis (JE) viruses in the forest in Asia, a seroepidemiological survey was carried out on 358 Southeast Asian cynomolgus (Macaca iris), 33 Indian bonnet (Macaca radiata) and 37 Japanese (Macaca fuscata) monkey sera by a plaque reduction neutralization test. The results indicated that Southeast Asian monkeys were naturally infected with these viruses but the frequency of antibody to them varied considerably according to the geographical origin of the monkeys. The frequency of antibody to one or more types of dengue virus were 87.2, 49.5, 34.3, 34.2 and 14.9% in Malaysian, Vietnamese, Cambodian, Indonesian and Filipino cynomolgus monkey sera, respectively. None of the Indian bonnet monkey sera neutralized type 1 dengue virus which was the only virus type examined with this monkey species. Monkey sera collected in Japan where dengue virus infection had not been known since 1944 did not significantly neutralize dengue viruses. JE virus antibody was detected at 29.7, 9.0, 8.6, 2.7, 1.4 and 0% in Japanese, Cambodian, Vietnamese, Indonesian, Filipino and Malaysian monkey sera respectively.

Animals↗
PubMed1991

Enzyme-linked immunosorbent assay for the detection of canine coronavirus and its antibody in dogs.

Two methods of enzyme-linked immunosorbent assay (ELISA) were developed for the diagnosis of canine coronavirus (CCV) infection in dogs. One ELISA, in which CCV-infected CRFK cell lysate is used as antigen, is for the detection and titration of antibody against CCV, and the other ELISA uses the double antibody sandwich method for the detection of CCV antigen. The first ELISA procedure demonstrated antibody responses in dogs inoculated with CCV, as did the virus neutralization test; the second ELISA detected specific CCV antigen in feces and organ homogenates of inoculated dogs.

Animals↗
PubMed1952

Studies on the propagation in vitro of poliomyelitis viruses. I. Viral multiplications in tissue cultures employing monkey and human testicular cells.

Poliomyelitis virus was propagated in vitro successfully in extraneural tissues. Suspended tissue fragment cultures and combined plasma clot-suspended tissue fragment cultures of monkey or human testicular tissues were employed. Five strains representative of poliomyelitis virus were maintained for from 36 to 263 days in the suspended tissue fragment type of culture. The dilution factors calculated by tissue replacements for the eight serial passages ranged from 10(7.8) to 10(44.5) and when assessed by fluid replacements, from 10(15) to 10(95.3). The LD(50) for each strain of Type 2 virus was determined for selected transfers. The identify of each strain of virus was established by neutralization tests and histopathological findings in monkeys dead from the injection of tissue culture virus. Control experiments and other tests made known that propagation of poliomyelitis virus did not occur in the absence of viable testicular cells and that an extraneous virus was not inadvertently acquired during the course of these studies.

Animals↗