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PubMed1991

Characterization of Sendai virus-induced human placental trophoblast interferons.

Human placental trophoblast cultures produce a mixture of interferons (IFNs) when challenged with Sendai virus. High-performance dye-ligand and immunoaffinity chromatography of a trophoblast IFN (tro-IFN) preparation enabled the isolation of three antigenically distinct IFNs, alpha I, alpha II 1 and beta, with Mrs of 16K, 22K and 24K respectively, by reducing and non-reducing SDS-PAGE. The major IFN, responsible for 75% of the total antiviral activity, was tro-IFN-beta, with the remaining activity being due to tro-IFN-alpha I and tro-IFN-alpha II 1, as determined by an antiviral neutralization test using specific anti-human IFN antibodies. The antiviral activities of the tro-IFNs were stable at pH 2.0 for 24 h and tro-IFN-alpha II 1 and -beta were shown to be glycoproteins. The three tro-IFNs showed different antiviral activities when assayed on human and bovine cell species; tro-IFN-alpha I and alpha II 1 protected both human and bovine (MDBK) cells from virus infection, whereas tro-IFN-beta showed a high degree of species specificity, protecting only the human cell types tested.

Animals↗
PubMed1983

Antigenic relationships among some bovine rotaviruses: serum neutralization and cross-protection in gnotobiotic calves.

A method was further developed to screen non-tissue-culture-adapted bovine rotaviruses for serotype, using a neutralization test with infectious fecal rotavirus. One of those rotaviruses (B223) which was not blocked by antiserum to the neonatal calf diarrhea virus (NCDV) serotype was then adapted to cell culture in the presence of the antiserum for two or more passages and hyperimmune antiserum to this isolate had a 60-fold-higher homologous neutralization titer than with the NCDV serotype rotavirus. Seventy-three isolates were serotyped and eight (11%) were not of the NCDV serotype (bovine rotavirus serotype I). Of these eight, five belonged to the new bovine rotavirus serotype II and three were not typed, indicating the existence of one or more further serotypes. Cross-protection studies in gnotobiotic calves showed that cross-protection only occurred between rotaviruses of the same serotype, and even a minor serotype difference was sufficient for the calves to show a lack of cross-protection. The serotypes (I and II and the three untyped isolates) also showed differences in the rate of migration in polyacrylamide gel electrophoresis of some of their RNA segments (no. 4, 6, 7, 8, 9, 10), indicating that they were of different electropherotypes.

Animals↗
PubMed1984

Characterization of dermonecrotic toxin produced by serotype D strains of Pasteurella multocida.

Dermonecrotic toxin (DNT) produced by serotype D strains of Pasteurella multocida, isolated from pigs, was characterized and compared with DNT produced by Bordetella bronchiseptica. The DNT prepared by sonication from P multocida or B bronchiseptica had dermonecrotic activity and lethal toxicity for guinea pigs and mice, and also induced marked atrophy of spleens in the mice. Toxicity of P multocida or B bronchiseptica DNT was completely inactivated by heating at 70 C for 30 minutes, and was reduced by treatment with trypsin, formalin, or glutaraldehyde, indicating that the DNT may be a protein. Although biologic and toxic properties of P multocida DNT were similar to those of B bronchiseptica DNT, cross-neutralization tests between P multocida and B bronchiseptica indicated that DNT from the 2 bacterial species were serologically distinct.

Animals↗
PubMedDate not supplied

Further studies on in vitro and in vivo assays of hemorrhagic enteritis virus (HEV).

Isolation of hemorrhagic enteritis virus (HEV) from spleens of infected turkeys in the MDTC-RP19 lymphoblastoid cell line was compared with detection of HEV antigen in the same spleens using the agar gel precipitation (AGP) test. A concordance of 80% was found between the two assays. Virus isolation had a sensitivity of 84% and a specificity of 88% compared with the AGP test. RP19 cells were also susceptible to infection with several other avian adenoviruses, but such infection was easily differentiated from that of HEV by a fluorescent-antibody (FA) test. Turkeys required 10(2) tissue-culture-infectious doses (TCID) to develop HE-specific lesions and 10(5) TCID to be killed. On the other hand, as little as 10 TCID of apathogenic HEV protected the poults against challenge with virulent HEV. The enzyme-linked immunosorbent assay (ELISA) for detection of HEV antibody was improved by using virus-infected RP19 cells as antigen. The ELISA appears to be more sensitive than the serum-neutralization test.

Animals↗
PubMedDate not supplied

Enzyme-linked immunosorbent assay for detection of antibody to avian encephalomyelitis virus in chickens.

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to avian encephalomyelitis virus (AEV) has been developed for determining whether existing AEV control programs adequately protect breeder hens. A partially purified AEV antigen was bound to microcuvettes for reaction with specific primary antibody. A second antibody, rabbit anti-chicken immunoglobulin G (IgG) conjugated with horseradish peroxidase, was employed to react with bound primary IgG. The relative amount of bound primary IgG was detected using ortho-phenylenediamine as a substrate for enzymatic production of a chromogen by horseradish peroxidase. Intensity of absorbance of the chromogen at 490 nm was related to the bound primary antibody by the titration method. Negative antisera were surveyed to establish an appropriate positive/negative cutoff level at twice the mean absorbance of negative sera at a 1:100 dilution. The test reagents for the ELISA were optimized by reagent titrations utilizing known positive and negative antisera for discrimination. The optimized ELISA had a coefficient of variation of from 1.2 to 3.3 for within-assay titer and of 2.4 for between-assay mean titer. Even though the ELISA detected only specific IgG, it was as accurate as the virus-neutralization test for evaluating the immune status of hens to AEV. Moreover, the ELISA was more economical in the use of reagents, time, and personnel and was free from dependence on susceptible embryos. Since ELISAs can be standardized and measured with manual or automated instruments, the derived ELISA can be easily and economically used to evaluate the immune status of breeder hens in commercial poultry operations.

Animals↗
PubMed1972

Rabies antibody determination by immunofluorescence in tissue culture.

A tissue culture (TC)-fluorescent antibody (FA) technique for the measurement of rabies-neutralizing antibody was found to be reliable and comparable to the standard mouse serum neutralization test. This test was performed with BHK-21 cells infected with the ERA vaccine virus strain on Lab-Tek TC chamber slides. A Flury high egg passage (HEP) rabies virus strain grown on continuous line of African green monkey (Vero) and on BHK-21 cells was investigated to determine its utilization in a TC-FA neutralization procedure. Although both the HEP and ERA viruses infected Vero and BHK-21 cells, the amount of fluorescent antigen observed was most consistent with ERA virus and BHK-21 cells.

Animals↗
PubMed1997

Puumala and Dobrava viruses cause hemorrhagic fever with renal syndrome in Bosnia-Herzegovina: evidence of highly cross-neutralizing antibody responses in early patient sera.

Hantavirus infection was diagnosed serologically by mu-capture IgM and IgG ELISAs in hemorrhagic fever with renal syndrome (HFRS) patients admitted to Tuzla Hospital, Bosnia-Herzegovina. The results indicated that more than one hantavirus caused the outbreak. To address the question of which hantavirus serotypes were involved, sequentially drawn sera were analyzed by focus reduction neutralization test (FRNT) for antibodies against Puumala, Hantaan, Dobrava, and Seoul hantaviruses. The data revealed that acute- or early convalescent-phase sera, even when drawn as late as 3 weeks after the onset of disease, could not be used for typing of the causative hantavirus; a significant number of these samples showed similar reactivity of neutralizing antibodies to several different hantavirus serotypes. Moreover, although several acute-phase sera showed the highest FRNT titer to Hantaan virus, convalescent sera from these patients in all cases showed high specificity for Puumala or Dobrava viruses. This phenomenon, interpreted as a cross-neutralizing primary antibody response, makes several earlier reports concerning causative agents of HFRS questionable. Serological examination of small rodents trapped in the endemic area identified Puumala- and Dobrava-like virus infections. RT-PCR and sequencing of rodent lung samples identified Dobrava virus in one yellow-necked field mouse (Apodemus flavicollis). Cross-FRNT data, using polyclonal rabbit antibodies, clearly confirmed Dobrava virus as a unique hantavirus serotype. In conclusion, the results revealed that both Puumala- and Dobrava-like viruses caused HFRS in Bosnia-Herzegovina, whereas no signs of Hantaan or Seoul virus involvement were found.

Animals↗
PubMed1982

Antibody responses to Epstein-Barr virus-specific DNase in relation to the prognosis of juvenile patients with nasopharyngeal carcinoma.

We have examined serial sera from 17 juvenile patients with nasopharyngeal carcinoma (NPC) for their capacity to neutralize the activity of Epstein-Barr virus (EBV)-specific DNase. The results revealed that NPC patients who became long-term survivors (LTS) without evidence of the disease either never possessed significant levels of antibodies to the enzyme or showed a gradual decline in the number of EBV DNase units neutralized from an elevated level at diagnosis to an insignificant figure several years later. All the 10 LTS neutralized less than 4, and some neutralized less than 2 units of the enzyme 3 or more years after the initial diagnosis. In contrast, serial sera from juvenile patients who died of NPC neutralized over 10 and as many as 25 units of EBV DNase either persistently until death occurred or with transient declines during unmaintained remissions. Rises and declines in the neutralizing activity were, with few exceptions, accompanied by corresponding changes in the titers of IgA and IgG antibodies to EB viral capsid antigen and to the diffuse component of the early antigens. Although the number of juvenile NPC cases available for study was small, the observations suggest that the EBV DNase neutralization test may serve to provide information on the prognosis of the patients.

Adolescent↗
PubMed1980

Parvovirus infection in translocated raccoons.

Severe enteritis attributed to parvovirus infection was diagnosed in raccoons (Procyon lotor) purchased from a commercial animal dealer for interstate translocation. Intestinal lesions included edema, hyperemia, and focal mucosal necrosis. Microscopically, intestinal villi were reduced markedly, and there was necrosis of the glandular crypts. Intranuclear inclusions were seen in many crypt cells. A parvovirus that was infective for Crandell feline kidney cells and primary canine kidney cells was isolated from the spleen of a sick raccoon. The raccoon virus was distinguished from feline panleukopenia virus by the former's ability to hemagglutinate swine erythrocytes at a pH of 7.2. Serum neutralization testing of infected raccoons indicated that the raccoon isolate differed from feline panleukopenia virus and may have been canine parvovirus.

Animals↗
PubMed2007

Isolation and identification of adenovirus from conjunctival scrapings over a two-year period (between 2001 and 2003) in Yokohama, Japan.

Over a 2-year period between 2001 and 2003, a total of 115 conjunctival scrapings were collected from patients with keratoconjuctivitis from several hospitals in Yokohama, Japan. Out of 115, 94 (82.4%) cases of adenoviruses were detected by polymerase chain reaction (PCR); 60 (52.1%) by cell culture isolation; and 16 (14.0%) by enzyme-linked immunosorbent assay (ELISA). The serotypes were determined by PCR- restriction fragment length polymorphism analysis (PCR-RFLP) and by the neutralization test (NT). PCR-RFLP was performed using a combination of endonucleases such as HhaI, AluI, and HaeIII. Of the 94 PCR-positive samples, the serotypes of 91 (96.8%) were identified by PCR-RFLP analysis (adenovirus 3: 50%, 4: 11%, and 8: 32%). Out of the 115 samples, 60 samples were identified by the neutralization (adenovirus 3, 4, 7, and 8). When both PCR-RFLP and the neutralization techniques were used, 53.2%, 11.7%, 1.1%, and 34% of the samples were identified as adenovirus 3, 4, 7, and 8, respectively. In contrast to the results of a nationwide surveillance report, adenovirus 3 was found as a major cause of keratoconjunctivitis in the Yokohama area. The nationwide surveillance report did not reflect accurately the epidemiological situation in the local area. In order to obtain surveillance data that would be useful for the prevention of an adenovirus conjunctivitis epidemic, it seems that local epidemiology is more important than that nationwide surveillance.

Adenovirus Infections, Human↗
PubMed2000

Antibodies to enteric adenoviruses (Ad40 and Ad41) in sera from Iranian children.

BACKGROUND: There is strong epidemiologic and serologic evidence that infection with the enteric adenoviruses can result in severe gastroenteritis in children. OBJECTIVES: This study was performed to determine the prevalence of enteric adenovirus infection in Iran. STUDY DESIGN: One hundred and twenty-seven single sera from children up to 7 years of age, collected from healthy Iranian children in 1993-1994, were tested for antibodies to enteric adenoviruses by neutralization tests. RESULT AND CONCLUSION: Antibodies to enteric adenoviruses have been detected in about one-half of sera. It is concluded that infection by these viruses is common among children in Iran.

Adenovirus Infections, Human↗
PubMed2004

TBE booster immunization in adults--first experience with a new tick-borne encephalitis (TBE) vaccine, free of protein-derived stabilizer.

A total of 222 adult subjects, all of whom received primary immunization according to the rapid immunization schedule in a preceding clinical trial with either a new (i.e. polygeline free) or formerly licensed (i.e. polygeline containing) TBE vaccine were invited for extension studies. The subjects received the first booster immunization with the new TBE vaccine at 12 to 18 months after primary immunization. Subsequently, a total of 191 of the 222 subjects could be enrolled in a serological follow-up one year after the booster immunization. Neutralizing TBE antibody titers were determined prior to, 21 days after and approximately 12 months after booster immunization. Prior to first booster immunization, TBE antibodies (GMTs) had remained on a high level and were far above the detection limit of the neutralization test used. All subjects of the per protocol population who were primarily immunized with the new TBE vaccine formulation and all but one subject of the control group were still seropositive prior to the booster. All subjects showed a sharp increase of TBE antibodies following the booster immunization. Within the 12 months follow-up period, neutralizing TBE antibody titers remained on a high level. The booster vaccination was well tolerated by the subjects. Only very few febrile reactions (< 1%) none higher than 38.5 degrees C were reported. No serious or unexpected adverse events related to vaccination were reported. These successful results in terms of both immunogenicity and safety indicate that TBE vaccination with this new TBE vaccine can be used safely in adults. A long lasting immunity can be concluded from the strong immune response following the booster immunization.

Adult↗
PubMed1984

Population studies of diphtheria immunity using antitoxin radioimmunoassay.

A quantitative method for testing serum diphtheria antitoxin levels was set up using a diphtheria antitoxin radioimmunoassay (RIA). The results of this RIA correlated well with the Schick test in 554 subjects and with intradermal neutralization tests in guinea-pigs in a small group of subjects. The RIA was suitable for use on blood collected by fingerprick on to a disc of standard chromatography paper. These discs could be stored at room temperature for at least 1 month. If storage for more than 6 months was required -20 degrees C was found to be better. Experience with this RIA in a total of 2349 subjects indicated that it is more accurate, rapid and less costly than Schick testing. The RIA should prove to be the preferred method for testing diphtheria immunity in population surveys.

Adolescent↗
PubMed2001

Serological status for Chlamydophila psittaci, Newcastle disease virus, avian polyoma virus, and Pacheco disease virus in scarlet macaws (Ara macao) kept in captivity in Costa Rica.

From 1998 to 1999, a total of 128 blood samples were collected from scarlet macaws (Ara macao), kept in captivity in 11 different aviaries located in six provinces of Costa Rica. The sera were examined for antibodies directed against Chlamydophila psittaci, Newcastle disease virus (NDV), avian polyoma virus (APV), and Pacheco disease virus (PDV). Testing by enzyme-linked immunosorbent assay (ELISA), showed 16 (12.39%) of the samples (n = 129) exhibited antibodies directed against C. psittaci. Employing haemagglutination inhibition tests for NDV antibodies, all of the samples were found to be negative. The prevalence of antibodies specific for APV was tested with a blocking ELISA and serum neutralization tests (SNT) and 12 of 128 samples (9.37%) were found to be positive with both tests. In SNT, two out of 128 samples (1.56%) were positive for PDV. This is the first description of the serological status in scarlet macaws in captivity in Costa Rica. The study demonstrates the absence of NDV antibodies in the birds investigated on one hand, but also indicates a health hazard for numerous avian species due to the risk of infections with C. psittaci, APV or PDV.

Animals↗
PubMed1993

Physician gender bias in clinical decisionmaking: screening for cancer in primary care.

There has been increasing interest in gender disparities in clinical decisionmaking. Few studies have examined this issue in nationally representative samples or focussed on primary care. In addition, few of the studies have examined the role of physician gender. The 1987 National Medical Expenditure Survey was used to examine the relationship between physician gender and screening deficiency in women for three gender-sensitive tests (breast examinations, Papanicolaou tests, and mammograms) and one gender-neutral test (blood pressure checks). Women reporting a female physician as their usual provider compared with those reporting a male physician were less likely to be deficient for Papanicolaou tests and mammograms. There was a smaller, but nonsignificant similar trend for breast examinations. No gender bias was evident for blood pressure checks. These results persisted after multivariate adjustment for patient age, race, education, income, insurance status, subjective health status, other health behaviors, and attitude toward health care and health insurance. The results confirm the existence of physician gender bias in clinical decision making and represent one area for quality improvement.

Adult↗
PubMed1981

Orbiviruses and bunyaviruses from a seabird colony in Scotland.

Viruses isolated from ticks (Ixodes uriae) and a kittiwake (Rissa tridactyla) from a seabird colony at St. Abb's Head, Scotland, were shown by complement fixation tests (CFT) to be antigenically related to the Uukuniemi and Kemerovo serogroups. Electron microscopic examination of cell cultures infected with the Kemerovo group viruses revealed particles characteristic of orbiviruses, 72 +/- 3 nm in diam., with an inner core 37 +/- 3 nm in diam., in association with intracytoplasmic, densely staining granular areas, and with fibrillar and tubular structures. Cell cultures infected with the Uukuniemi group viruses revealed characteristic bunyavirus particles, 94 +/- 7 nm in diam., with a closely adherent envelope. Both orbi- and bunyaviruses were isolated from two tick pools and the kittiwake. A third tick pool contained an orbivirus which cross-reacted with the other isolates in CFT and fluorescent antibody tests, but was distinguished from them by neutralization tests.

Animals↗
PubMed2006

Development of serological assays for Thottapalayam virus, an insectivore-borne Hantavirus.

Thottapalayam virus (TPMV), a member of the genus Hantavirus in the family Bunyaviridae, was isolated from an insectivore, Suncus murinus (musk shrew), captured in southern India in 1964. While the isolation of TPMV predates the discovery of the prototype Hantaan virus, little is known about its genetics and biology. To date, preliminary evidence suggests that TPMV differs significantly, both antigenically and genetically, from all known rodent-borne hantaviruses. However, since detailed epizootiological studies have not been conducted, it is unclear if TPMV is naturally harbored by an insectivore host or if TPMV represents a "spillover" from its natural rodent reservoir host. Moreover, to what extent TPMV causes infection and/or disease in humans is not known. To address these issues, we first studied the antigenic profile of TPMV using monoclonal antibodies against Hantaan and Seoul viruses and polyclonal immune sera against Puumala virus and TPMV. Armed with this newfound information, we developed an enzyme-linked immunosorbent assay system for the diagnosis of TPMV infections in shrews and humans, using a recombinant TPMV N antigen manipulated to have an E5/G6 epitope to be captured by monoclonal antibody clone E5/G6. Using this assay, we found anti-TPMV antibodies in sera from a patient with high fever of unknown etiology in Thailand and from two shrews captured in Indonesia. Seropositivity was verified by the indirect immunofluorescence antibody test, Western blotting analysis, and focus reduction neutralization test. Collectively, our data indicate that TPMV is harbored by Suncus murinus as its host in nature and is capable of infecting humans.

Animals↗