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At least 1,135 records · Page 63Linked to original sources
PubMed1981

Possible "immuno-protection" of the bovine parvovirus in the uterus: Preliminary communication.

A pregnant Holstein cow was experimentally inoculated with bovine parvovirus. Approximately five weeks after the last of three injections, the animal aborted a fetus which showed signs of mummification. Virus particles similar to bovine parvovirus were identified by electron microscopy from fetal lung tissue and intestinal contents. Five months after the abortion, the experimental animal died of a corynebacterial metritis and septicemia. Immunofluorescence tests were performed on representative tissues obtained at necropsy. Of all tissues examined only the uterus showed a positive response to the immunofluorescence assay, indicating that the viral antigen persisted in the uterine environment long after its elimination from the systemic circulation.

Journal Article↗
PubMed1985

An enzyme immunoassay for identification and quantification of infectious murine parvovirus in cultured cells.

An enzyme immunoassay was developed to identify and quantify infectious murine parvovirus. The assay was as sensitive as the fluorescent focus test for quantification of virus in clinical material and was less labor-intensive. Viral infectivity could be inactivated with formalin prior to performance of the enzyme immunoassay. Specificity of the reaction product was shown by neutralization of infectivity using a hyperimmune rabbit antiserum to murine parvovirus.

Animals↗
PubMed1988

A latex agglutination test for the detection of canine parvovirus and corresponding antibodies.

Latex particles coated with rat monoclonal antibodies directed against a canine parvovirus are agglutinated by the virus antigens. In this study, the reaction was quantitated by a technique which had been described as immunoassay by particles counting. The method is as sensitive as a radioimmunoassay with the same antibody, but has the advantages of simplicity and safety. It allows the detection of parvoviral antigens to 4 ng/ml. Incubation of these antigens with specific antibodies resulted in inhibition of the agglutination reaction. By this procedure the presence of antibodies against canine parvovirus can be detected to a concentration of 150 ng/ml. The system can be easily automated, thus increasing reproducibility. This latex agglutination technique can be performed as a slide test; and although this method is 4-5 times less sensitive, it could be useful in field studies, for the detection of the viral antigens and the corresponding antibodies.

Animals↗
PubMed1990

Detection of B19 parvovirus infections by a dot-blot hybridization assay using a digoxigenin-labelled probe.

A non-radioactive dot-blot hybridization assay for the detection of B19 parvovirus infections was developed using a digoxigenin-labelled probe both on nylon and nitrocellulose filters. A 700 bp BamHI HindIII fragment of B19 DNA was used to construct the probe. Probe labelling was carried out by incorporating deoxyuridine triphosphate labelled with digoxigenin. The dot-blot hybridization assay was visualized by an immunoenzymatic reaction using antidigoxigenin Fab fragments labelled with alkaline phosphatase. The specificity and sensitivity of digoxigenin-labelled B19 DNA probe was compared with the results obtained with 32P-labelled B19 DNA probe. Out of the 504 serum samples tested, 3 samples were positive in all the hybridization assays performed and 494 were negative, 7 serum samples gave a weak positive reaction when Dig-B19 probe was used on nitrocellulose filters. The 77 pharyngeal swabs tested were negative in all the hybridization assays performed. Our hybridization assay showed a high sensitivity and reproducibility and it appears to be a rapid, practical and reliable test for routine screening of B19 parvovirus DNA in large numbers of clinical specimens.

Adolescent↗
PubMed1991

Polymerase chain reaction (PCR) amplification for the detection of porcine parvovirus.

A polymerase chain reaction (PCR) amplification method was developed and evaluated to detect porcine parvovirus (PPV). A pair of 20-base primers and an oligonucleotide probe were derived from the DNA sequences common to two isolates of PPV, NADL-8 and NADL-2. The primers flanked 118-bp nucleotides within the region coding for the major structural protein VP2. After DNA amplification of PPV replicative form (RF), a 158-bp fragment was detected in agarose gels. This amplified fragment was shown to be specific for PPV DNA after Southern transfer and hybridization to a 20-base internal probe. The amplified fragment also contained a single EcoRI cleavage site. Various conditions, such as number of cycles and annealing temperature, were examined to optimize the conditions for detecting viral DNAs from infected cell cultures and swine fetal tissues. Four different isolates of PPV, NADL-8, NADL-2, KBSH and Kresse, and two other viruses, canine parvovirus (CPV) and pseudorabies virus (PRV), were included to determine specificity of amplification. Slot blot hybridization with a radiolabeled probe was used to evaluate the sensitivity of PCR amplification. The optimized protocol was specific for PPV detecting equally all four strains of PPV, but failing to amplify CPV or PRV sequences. The PCR method could detect at least 100 fg of viral replicative form (RF) DNA or the equivalent of 1 PFU of infectious virus. The applications of this method include routine detection of PPV in clinical samples and as a contaminant in mammalian cell lines.

Animals↗
PubMed1989

Nucleotide sequence of capsid protein gene of porcine parvovirus.

Approximately 90% of the genome of porcine parvovirus was cloned into bacterial cells. The nucleotide sequence of the genome from 33 map units (MU) to 95 MU was determined and shown to include the entire gene encoding the capsid proteins. The predicted amino acid sequences of the capsid proteins showed extensive homology to those of other autonomous parvoviruses.

Amino Acid Sequence↗
PubMed1990

Persistence of parvovirus H-1 DNA in human B- and T-lymphoma cells.

Persisting DNA of parvovirus H-1 could be demonstrated in cells of two human lymphoma cell lines, the Burkitt lymphoma cell line BL2 and the T-cell leukemia cell line Jurkat which survived infection with parvovirus H-1. Persistence of H-1 DNA rendered the cells resistant to a second H-1 infection. This resistance to H-1 superinfection persisted even after loss of H-1 DNA occurring after approximately 150-200 cell generations. Resistance to H-1 superinfection was accompanied by reduced uptake of infectious particles and by a block of H-1 DNA replication. This suggests that persistent H-1 infection leads to modifications of cellular functions involved in the permissivity for H-1.

Burkitt Lymphoma↗
PubMed1982

Studies on the multiplication of a porcine parvovirus.

A porcine parvovirus has been characterized with regard to its replication in foetal porcine kidney cells and certain biophysical properties. Electron microscopy of infected cells at selected times postinfection revealed that porcine parvovirus replication took place within or near a series of granular intranuclear inclusions which may be contiguous with cellular heterochromatin. Developing virions were observed to aggregate into a nucleolar-like amorphous mass which gradually disrupted as cellular integrity was lost. Purified virions were found to have a buoyant density in CsCl of 1.38 g/ml, while 'empty' particles had a buoyant density of 1.29 g/ml. The particle diameter was calculated to be approximately 22 nm.

Animals↗
PubMed1985

Studies on the replication of a bovine parvovirus.

Optimal replication of a bovine parvovirus type 1 was found to occur when parasynchronous bovine embryonic lung cells were infected during the S phase of the cell cycle, just prior to maximum DNA synthesis. Viral antigen was first detected in the cytoplasm by immunofluorescence at 8 h post-infection, reaching a maximum at this location by 16 h and then disappearing. In the nucleus, antigen was first detected at 12 h, concurrent with early inclusion body formation and first detection of intracellular virus production. Intranuclear antigen then increased rapidly to a maximum at 20 h, as the inclusions progressively matured, large amounts of virus were produced within the cell, with some release to the environment. From 24 h, the nuclear inclusions became increasingly shrunken and basophilic as virus migrated to the cytoplasm and was progressively released to the exterior concurrent with cell degeneration and fragmentation. The majority of virus remained cell associated, even at 28 h post-inoculation. Two morphological types of early and late stage intranuclear inclusions were produced by the virus, these appearing to be a distinct feature of bovine strains. In other aspects, the replication of bovine parvovirus appeared similar to that of other members of the genus.

Animals↗
PubMed1988

Mapping of porcine parvovirus DNA and development of a diagnostic DNA probe.

Dimeric and monomeric replicative forms of DNA of porcine parvovirus (PPV) strain NADL-2 were isolated and examined by restriction enzyme analysis and reciprocal Southern blot hybridization during development of a DNA probe for PPV. Genomic single stranded PPV DNA was 5.0 kb long, and results substantiated the rolling-hairpin model of parvovirus DNA replication with the primer sequence located in the 3' terminal hairpin loop. An additional finding was the generation of a 4.7 kb species of viral DNA which was considered to be a 0.3 kb deletion variant of genomic PPV DNA. A 3.0 kb DNA fragment obtained by Pst I/Hind III digestion of monomer replicative form DNA was cloned into a plasmid vector, pUC 19. The cloned fragment, recovered from transformed Escherichia coli strain TB1 and labelled with [32P] dCTP, was evaluated by dot hybridization as a probe for PPV in infected cell cultures. The probe was specific for PPV infected cells, and was 100 times more sensitive than the standard hemagglutination test.

Animals↗
PubMed1995

Development of an IgM capture assay for the diagnosis of B19 parvovirus infection using recombinant baculoviruses expressing VP1 or VP2 antigens.

BACKGROUND: The clinical manifestations of human parvovirus B19 infection are often similar to those induced as the result of infection by other infectious agents such as rubella and some bacteria. Although diagnosis of B19 infection is feasible by detection of specific antibodies, the tests require viraemic serum as a source of antigen. This inevitably leads to problems of reproducibility and dependence upon appropriate high quality clinical material. OBJECTIVES: To develop a monoclonal antibody capture ELISA (MACEIA) for detecting anti-B19 IgM antibody in human sera, using recombinant baculoviruses expressing the B19 parvovirus VP1 and VP2 proteins and to compare this with MACEIA using a plasma derived B19 antigen. STUDY DESIGN: Sera from 85 patients with proven B19 infection and the paired convalescent sera from 26 anti-B19 IgM-positive acute samples were examined for B19-specific IgM antibody by a monoclonal antibody capture assay that utilised recombinant baculoviruses expressing B19 proteins in lieu of a plasma-derived B19 antigen. Control samples consisted of 24 anti-rubella IgM, 24 anti-EBV IgM and 102 negative sera from uninfected individuals. RESULTS: Eighty-four of the 85 sera were anti-B19 IgM positive by MACEIA using recombinant baculovirus derived B19 antigen and by indirect immunofluorescence tests, whereas 79 were positive by MACEIA using plasma-derived antigen. Of the 26 convalescent samples which were positive as acute sera, 4 had become negative by 8 weeks post-infection. The expressed recombinant baculovirus antigens had identical molecular weights to the VP1 (84 kDa) and VP2 (58 kDa) proteins of virus purified from human plasma. Recombinant baculovirus-derived VP1 antigen was as effective as VP2 particles at detecting antibodies. CONCLUSIONS: Recombinant proteins VP1 and VP2, obtained from recombinant baculovirus-infected cell lysate, showed equal specificity to and higher sensitivity than, B19 virus purified from human plasma when used in MACEIA to detect B19-IgM antibody.

Journal Article↗
PubMed1996

IgG subclass response to human parvovirus B19 infection.

BACKGROUND: IgG antibodies are essential to immunity against human parvovirus B19 and can neutralize infection both in bone marrow cell cultures infected in vitro and in chronically infected immunosuppressed individuals. OBJECTIVES: To assess the levels and response kinetics of IgG subclasses towards individual structural proteins of human parvovirus B19. STUDY DESIGN: Subclasses of IgG for capsid proteins VP1 or VP2 were quantified by EIA using monoclonal antibodies in 30 acutely infected and 30 convalescent patients, as well as in 32 remotely infected and 20 non-infected controls. RESULTS: In all groups of seropositive individuals the predominant subclass for either structural protein was IgG1. Subclass IgG3 was associated with acute infection. By contrast, IgG4 appeared months after infection, and occurred specifically towards VP1. The ratio of VP1-specific subclasses IgG3 and IgG4 provided a diagnostic test for recent infection with a specificity of 98% and a sensitivity of 97%. CONCLUSIONS: Comparative measurement of VP1-specific IgG3 and IgG4 is useful in diagnosis. The IgG4 results point to long-term expression of immunologically active VP1 and to T-cell help of T(h)2 type for B-cells recognizing VP1.

Journal Article↗
PubMed2006

Parvovirus B19 reactivation presenting as neutropenia after rituximab treatment.

A patient with primary biliary cirrhosis and associated refractory immune thrombocytopenic purpura was treated with 4 weekly courses of rituximab, a monoclonal antibody targeting B-cell surface antigen CD20. Her thrombocyte count and even cholestatic liver function tests improved. However, 17 weeks after rituximab treatment, she developed severe neutropenia (absolute neutrophil count 0.23x10(3)/mul) and recurrent thrombocytopenia with abnormal bone marrow of all three lineages. Although delayed-onset neutropenia has been reported after rituximab, reactivated viral infections have also been encountered. Parvovirus B19 was suspected and confirmed as the cause of neutropenia in our patient. The patient was supported by GCSF treatment and recovered uneventfully after several weeks. Neutropenia after rituximab can also be the predominant manifestation of reactivated parvovirus B19 infection and have a favorable prognosis.

Journal Article↗
PubMed1995

Transcriptional inhibition of the parvovirus minute virus of mice by constitutive expression of an antisense RNA targeted against the NS-1 transactivator protein.

We have assessed a genetic resistance approach based on antisense RNA to interfere with the prototype Minute Virus of Mice (MVMp), an autonomous parvovirus. MVMp is a cytolytic virus when infecting the permissive A9 mouse fibroblast cell line, and its gene expression is largely regulated at the level of transcription initiation by the nonstructural transactivator NS-1 protein, a multifunctional polypeptide also involved in viral DNA replication and cytotoxicity. An NS-1 specific antisense RNA constitutively expressed in transfected A9 clones increased several fold the proliferative viability of the cells upon high multiplicity virus infection, and cultures infected at low multiplicity reached confluence overcoming virus progression. All clones shared a common phenotype of resistance characterized by a lowered synthesis of viral DNA replicative intermediates and genomic forms, a significant reduction in the accumulation of the three viral messengers in the cytoplasmic and nuclear compartments, and a specific inhibition in viral protein synthesis. These results indicate that the constitutive antisense RNA mediates an overall repression of viral macromolecular synthesis by preventing the onset of NS-1 functions. Therefore, cytocidal parvoviruses may be hampered by engineered antisense RNA targeted against early regulators of virus growth.

Animals↗
PubMed1988

Fetal infection with human parvovirus B19.

The case of premature infant with nonimmune hydrops secondary to human parvovirus B19 infection is presented. The infection was not suspected clinically. The fetal autopsy showed intranuclear inclusions in erythroid cells that stained positively with a DNA hybridization probe for B19 parvovirus.

Edema↗
PubMed1981

Outbreak of aplastic crises in sickle cell anaemia associated with parvovirus-like agent.

Since 1952, 112 children with sickle cell anaemia (SCA) in Jamaica have had an aplastic crisis. Outbreaks occurred in 1956, 1960, 1065-67, 1971-73, and 1979-80. Most cases occurred in children under 10 years of age, and an aplastic crisis in a patient over the age of 15 years is rare. There were 38 cases in 1979-80 and stored serum specimens from 28 of these were available for virus studies. Evidence for infection with a parvovirus-like agent was found in 24 of these 28 cases. Viral antigen was detected in 2 patients, both of whom demonstrated seroconversion. Seroconversion during 1980 was detected in a further 7, increasing amounts of antibody during the convalescent period were found in 5, antibody was found in 2 of 4 patients from whom only an acute phase specimen was available and the remaining 10 were antibody positive in the only convalescent phase sample available for testing. Antibody was found in 4 of 94 controls with the SS genotype (in retrospect 2 of these may have had an aplastic crisis) and in 17% of 48 controls with a normal haemoglobin (AA) genotype. The results accord with the possibility that the parvovirus-like agent is the principal cause of aplastic crisis in SCA.

Adolescent↗
PubMed1983

Aplastic crisis due to parvovirus infection in pyruvate kinase deficiency.

A thirteen-year-old boy with congenital haemolytic anaemia due to pyruvate kinase (PK) deficiency had an aplastic crisis. A serum parvovirus-like virus (SPLV) was demonstrated in the blood by electron microscopy and, subsequently, IgM and IgG antibodies to the prototype SPLV B19 were detected. In an attempt to define the level of erythropoiesis that is involved in parvovirus-induced bone marrow suppression, the levels of circulating early erythroid progenitors (burst forming units erythroid, BFU-E) were monitored during the crisis and recovery period. The virus-containing plasma inhibited the formation of BFU-Es from non-immune subjects and this effect was neutralised by convalescent serum. Colony forming units granulocyte-macrophage (CFU-GM) were also inhibited but this was probably non-specific since neutralisation did not reverse the effect. These experiments, together with the clinical data, suggest a selective effect of SPLV at the stage of erythroid progenitors.

Adolescent↗