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PubMed2004

Evaluation of the Roche LightCycler parvovirus B19 quantification kit for the diagnosis of parvovirus B19 infections.

BACKGROUND: The rapid and quantitative detection of viral DNA is important in the diagnosis of parvovirus B19 infection in immunocompromised patients and in congenital infection. It is also valuable for monitoring progress following therapeutic interventions. OBJECTIVES: To evaluate the diagnostic sensitivity and specificity of the Roche LightCycler (LC) parvovirus B19 quantification kit in comparison with previously described nested PCR and dot blot hybridisation assays. STUDY DESIGN: Two hundred and twenty eight clinical samples and two standard B19 DNA sera were tested to assess the diagnositic performance of the Roche LC kit. RESULTS: Ten clinical samples (4.3%) gave invalid LC results, including three of five bone marrow samples but only two of 165 serum samples. In the remaining 218 samples, the LC assay detected B19 DNA in 97.5% (79/81) samples that were positive by the nested PCR. The two samples (from the same patient) that were LC negative were sequenced in a 511-nucleotide region of the NS gene and 42 nucleotide changes were found. The Roche LC assay detected B19 DNA in 9.5% (13/137) samples that were negative by nested PCR. Analysis of the available clinical and serological data associated with these samples suggested that the LC results in the majority of these cases were true positive. In patients with resolving persistent infection, the LC assay remained positive for longer than nested PCR. CONCLUSIONS: The Roche LC assay was more sensitive than the nested PCR used in this study. The additional sensitivity and the quantitative DNA measurements were valuable for monitoring patients with persistent B19 infection. Practical advantages of the LC assay include a short running time and the possibility to automate the assay. The LC assay provides a controlled and standardised method for quantitative detection of viral DNA for the diagnosis and monitoring of parvovirus B19 infections but failed to detect a variant strain.

Adult↗
PubMed1997

Epidemiological characterization of newly recognized rat parvovirus, "rat orphan parvovirus".

Newly recognized rat parvovirus (rat orphan parvovirus: ROPV) was examined for viral excretion and persistence in infected rats, and also for infectivity to mice and hamsters. The virus appeared to replicate mainly in lymphoid or hematopoietic tissues, and was detected in feces, urine and oropharynx of the infected rats at 1 to 4 weeks postinfection. The infective virus was also detected in peripheral leukocytes and various tissues at an acute phase of infection, and decreased in every tissue at 8 weeks postinfection. Viral DNA, however, was persistent in lymphoid tissues at least up to 24 weeks postinfection. When the virus was inoculated to mice and hamsters, no evidence of viral production and antibody response was demonstrated. ROPV is assumed to be a variant of the known rat parvovirus which resulted to alter cell tropism and persist in lymphoid or hematopoietic tissues, in order to escape from host immune system.

Animals↗
PubMed1984

[Parvovirus (serum-parvovirus-like-virus) and Aurillac antigen. A study of 18 cases].

Two cases of parvovirus viraemia were observed in 1972 by screening of blood donors for HBs Ag by counter-electrophoresis. This virus was named "Aurillac" from the town in which the first case originated. In London, a virus found in the same circumstances in 1975, was found identical to the Aurillac Ag. The authors named it "serum parvovirus-like virus" (SPLV) or parvovirus B 19. Since 1972, 16 additional cases have been observed in Paris. Among these 18 cases, 11 were blood donors, 4 were hospital staff and 3 were patients. Second serum samples were available for 13 of the 18 cases. Seroconversion has been detected in all except one, a patient with lymphosarcoma. Antibody could be observed within few days. Anti-Aurillac antibody was found by CEP in 25,3% of blood donors. Examination of antigen positive sera by immune electron microscopy using a late serum sample showed aggregates of spherical particles measuring 20 nm in diameter. The antigen banded in caesium chloride at a density of 1.38. Ten of the 15 healthy adults could be interviewed: 7 appeared symptomless, 1 had a fever up to 39 degrees C for one day, 2 had erythema (as well as 3 subjects who had been in contact with these 2 persons). These cases are discussed and compared to the British literature showing that this virus could provoke the fifth disease or erythema infectiosum and aplastic crisis in chronic hemolytic syndromes. This virus could contaminate clotting factors.

Adolescent↗
PubMed1987

Detection of bovine parvovirus proteins homologous to the nonstructural NS-1 proteins of other autonomous parvoviruses.

Two nonstructural proteins of bovine parvovirus (BPV) with apparent molecular sizes of 75,000 and 83,000 daltons have been detected. The proteins were immunoprecipitated from lung cells infected with various isolates of BPV and from in vitro translations of infected cell mRNA. These proteins were expressed as nuclear phosphoproteins and were synthesized early in infection, before the peak of capsid protein synthesis. Early in infection, the 75-kilodalton-size species could be resolved into two bands of equal intensity, but later in infection, the lower-molecular-size form predominated. Antibodies directed against bacterial fusion proteins encoding amino acid sequences from a highly conserved region of the NS-1 polypeptides of two other parvoviruses, minute virus of mice and the human virus B19, gave specific nuclear fluorescence with BPV-infected cells, although the antibodies failed to immunoprecipitate any viral proteins. The noncapsid proteins appear to be homologous to the previously characterized NS-1 proteins of other autonomous parvoviruses.

Animals↗
PubMed1987

Comparisons of feline panleukopenia virus, canine parvovirus, raccoon parvovirus, and mink enteritis virus and their pathogenicity for mink and ferrets.

Parvoviruses from mink (mink enteritis virus [MEV]), cats (feline panleukopenia virus [FPV]), raccoons (raccoon parvovirus [RPV]), and dogs (canine parvovirus [CPV]) were compared. Restriction enzyme analysis of the viral replicative-form DNA revealed no consistent differences between FPV and RPV isolates, but CPV and MEV isolates could be distinguished readily from other virus types. Feline panleukopenia virus, RPV, and MEV, but not CPV, replicated to high titers in mink. However, on the first passage, disease and microscopic lesions were observed only in mink inoculated with MEV. Feline panleukopenia virus and RPV isolates replicated in ferrets, but disease or microscopic lesions were not observed. Feline panleukopenia virus and RPV isolates could be passaged repeatedly in mink and ferrets. Virulence of FPV and RPV isolates was low compared with that of MEV, and only a single mink inoculated with FPV or with RPV developed clinical disease on the sixth passage of virus.

Animals↗
PubMed2002

Parvovirus B19 and parvovirus V9 are not associated with Henoch-Schönlein purpura in children.

BACKGROUND: Based on single case reports, parvovirus B19 (B19) has repeatedly been proposed as an etiologic agent in patients with Henoch-Schönlein purpura (HSP), perhaps causing vasculitis by direct invasion of vascular endothelial cells because of the tissue distribution of the cellular B19 receptor. A cohort of children with HSP and other vasculitic diseases was investigated and compared with healthy control children to assess the role of B19 as well as parvovirus V9 (a putative emerging B19-like virus). PATIENTS AND METHODS: Serum samples from 36 children with HSP (n = 29) or other vasculitic diseases (n = 7) were examined, and 38 healthy bone marrow donors were used as controls. The presence of specific B19 and V9 IgM and IgG antibodies was determined with a recently developed enzyme-linked immunosorbent assay, and viral DNA was detected by a novel nested PCR. RESULTS: Specific IgM was not present in any of the patient or control serum samples. B19 DNA was detected in one patient, a previously healthy 8-year-old boy diagnosed with HSP, whereas none of the controls was B19-positive. V9 was not detected in any of the clinical or control samples. It seems likely that B19 infection might have triggered the development of HSP in the B19-positive patient, because B19 viremia is otherwise uncommon. CONCLUSIONS: Although causality is difficult to construe in single cases, the data indicate that B19 is not a common contributing factor in the pathogenesis of vasculitis and that this pathogen is only rarely associated temporally with HSP or vasculitic diseases in children.

Adolescent↗
PubMed2002

The canine minute virus (minute virus of canines) is a distinct parvovirus that is most similar to bovine parvovirus.

We characterized the genome and proteins of the canine minute virus (the minute virus of canines (MVC)). The genome sequence showed MVC to be an autonomous parvovirus encoding a large nonstructural protein 1 gene, a smaller nonstructural protein, and overlapping VP1 and VP2 protein genes. The virus was most closely related to bovine parvovirus (BPV), with which it was 43% identical at the DNA sequence level, while the NS1 and VP1 proteins were 33.6 and 41.4% identical to those of BPV, respectively. Spliced messages of the NS1 gene transcripts were detected by RT-PCR. VP1 and VP2 proteins were detected in purified capsids, as were modified versions of each protein, and VP3 was also found in full capsids.

Amino Acid Sequence↗
PubMed2004

Effect of porcine parvovirus vaccination on the development of PMWS in segregated early weaned pigs coinfected with type 2 porcine circovirus and porcine parvovirus.

The objectives of this study were to determine if coinfection of segregated early weaned (SEW) pigs with porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV) induces an increase in the incidence of post-weaning multisystemic wasting syndrome (PMWS) compared to singular PCV2 infection, and to determine if vaccination against PPV protects pigs against PMWS associated with PCV2/PPV coinfection in SEW pigs. Seventy, 3-week-old, SEW pigs were randomly assigned to one of the five groups. Pigs in group 1 (n = 14) served as the negative controls, group 2 pigs (n = 14) were inoculated with PCV2, group 3 pigs (n = 12) were inoculated with PPV, groups 4 (n = 16) and 5 (n = 14) pigs were inoculated with both PCV2 and PPV. Pigs in groups 1-3 and 5 were vaccinated with two doses of a killed parvovirus-leptospira-erysipelothrix (PLE) vaccine prior to inoculation. The PCV2/PPV-coinfected pigs (groups 4 and 5) had significantly (P < 0.05) higher and more persistent fevers than the singular PCV2-infected pigs. One pig in each of the coinfected groups developed clinical disease (fever, respiratory disease, jaundice, weight loss) consistent with PMWS. Lymphoid depletion was significantly (P < 0.05) more severe in the dually-infected pigs at 42 days post-inoculation (DPI). Vaccinated, coinfected pigs (group 5) remained viremic significantly (P < 0.05) longer and had higher copy numbers of genomic PCV2 DNA in sera at 28, 35, and 42 DPI compared to the unvaccinated coinfected pigs (group 4). PPV-viremia was detected only in the unvaccinated group 4 pigs. PLE-vaccination prevented PPV-viremia but did not prevent clinical PMWS or reduce the severity of lymphoid depletion in PCV2/PPV-coinfected pigs. Evidence of increased incidence of clinical PMWS due to vaccination was not observed in this model.

Animals↗
PubMed1993

Electron microscopic identification of parvovirus virions in erythroid and granulocytic-line cells in a patient with human parvovirus B19 induced pancytopenia.

In this short report we describe a patient with human parvovirus B19 (HPV B19)-induced transient pancytopenia. Parvovirus virions were seen by electron microscopy in both erythroid and granulocytic precursors. Erythroid cells are not the only targets in these cases. We draw attention to this disorder so that physicians involved with hematological disorders and transplantation be more aware of this infection.

Adult↗
PubMed1992

Partial DNA cloning and sequencing of a canine parvovirus vaccine strain: application of nucleic acid hybridization to the diagnosis of canine parvovirus disease.

The cloning and sequencing of an Eco RI-PstI fragment derived from the replicative form of a canine parvovirus (CPV) vaccine strain are reported. The variability of the 5' end of NS 1 protein gene in the genome is confirmed by comparison with previously determined DNA sequences. A 15 nucleotide deletion was also observed in this vaccine strain. In order to improve CPV diagnosis, radioactively labelled RNA or DNA and biotin labelled DNA obtained by random priming of the recombinant plasmid were used as probes mainly on gut or stool samples from naturally infected dogs. Results of filter hybridization correlated well with histopathological diagnosis of parvovirus infection and with hemagglutination tests performed on dog faeces. We propose that nucleic acid hybridization may be an alternative diagnostic method to ascertain the presence of CPV, especially in frozen samples.

Amino Acid Sequence↗
PubMed1988

Canine host range and a specific epitope map along with variant sequences in the capsid protein gene of canine parvovirus and related feline, mink, and raccoon parvoviruses.

Canine parvovirus (CPV) is a recently recognized pathogen of dogs that is similar to the long-recognized feline, mink, and raccoon parvoviruses. Relationships between the viruses determined from DNA sequences of the capsid protein genes of 10 virus isolates showed the CPV isolates to be closely related to the other viruses, although comprising a distinct group. No immediate ancestor of CPV was observed amongst the mink, cat, or raccoon viruses examined. Three different directly repeated sequences were present within the noncoding region downstream from the capsid protein genes. Analysis of recombinants between CPV and feline panleukopenia virus at restriction sites within the capsid protein genes mapped a CPV-specific neutralization epitope on the virus capsid, differences in the pH dependence of hemagglutination, and part of the determinant of canine host range between 59 and 64 genome map units (m.u.). Those differences were therefore the result of up to three nucleotide or predicted amino acid sequence differences in that region. A second region between 64 and 73 m.u., which may affect the viability of certain recombinant viruses, contained four nucleotide differences, one of which was a coding change.

Animals↗
PubMed1983

Distribution of restriction enzyme sites in the bovine parvovirus genome and comparison to other autonomous parvoviruses.

A physical map has been constructed for bovine parvovirus (BPV) replicative form (RF) DNA synthesized in vitro. Analysis of restriction enzyme digestion products for 12 enzymes by neutral gel electrophoresis allowed the mapping of 26 cleavage sites between the 3' and 5' termini of the in vitro RF DNA. Cleavage sites for these enzymes were not detected within the 3'-terminal hairpin. HindIII, SalI, SmaI, SstII and XorII did not cleave BPV DNA, while 12 other enzymes cleaved the genome at one to four sites. Eleven additional enzymes were shown to cleave the genome at 8 to 20 sites. Comparison of the restriction site locations for BPV with those of other autonomous parvoviruses suggests that there may be conservation within the genome of certain restriction sites.

Animals↗
PubMed1994

Production of mink enteritis parvovirus empty capsids by expression in a baculovirus vector system: a recombinant vaccine for mink enteritis parvovirus in mink.

The VP-2 gene of mink enteritis parvovirus (MEV) was amplified by the polymerase chain reaction using MEV DNA isolated from the faeces of a naturally infected mink. Subsequently the VP-2 gene was cloned into a baculovirus expression vector. Recombinant baculoviruses were isolated and the MEV VP-2 gene product was characterized after expression in Sf9 insect cells. The MEV VP-2 product had the same size as that reported for the wild-type MEV VP-2 protein and was recognized by convalescent sera from MEV-infected mink and a panel of monoclonal antibodies reactive to MEV. Furthermore, the VP-2 protein was able to form parvovirus-like particles, which had haemagglutinating properties comparable with the wild-type MEV. The cloned VP-2 gene was sequenced and only five nucleotide differences were found after alignment with the known sequences of the MEV type 1 and type 2 isolates. Surprisingly, the VP-2 gene encoded a valine and a tyrosine at amino acid positions 232 and 234, identical to the situation found in MEV type 1, but at position 300 there was a valine which is a determinant of MEV type 2. Immunization of mink with approximately 40,000 haemagglutinating units of recombinant MEV VP-2 induced a measurable antibody response as tested by haemagglutination inhibition. Furthermore, the immunized mink did not excrete virus and did not develop clinical disease upon challenge with a virulent isolate of MEV.

Animals↗
PubMed1986

Latex agglutination test for detecting feline panleukopenia virus, canine parvovirus, and parvoviruses of fur animals.

A latex agglutination (LA) test for the detection of parvoviruses of fur animals, cats, and dogs was developed, and its sensitivity and specificity were compared with those of hemagglutination (HA) and the enzyme-linked immunosorbent assay (ELISA). Tissue culture isolation was used to confirm the specificity results. Fecal samples from various sources were tested, including specimens from raccoon dogs and mink which were experimentally infected with parvoviruses by oral exposure. LA compared favorably with the other tests. The ELISA was the most sensitive. When it was considered as a reference test, the corresponding sensitivities for HA and LA were 96 and 91%, respectively. The specificities were 93% for the ELISA, 95% for the HA test, and 92% for the LA test. LA seems to be a suitable technique for screening animals in the field and in laboratories in which sophisticated techniques are not available.

Animals↗
PubMed1984

Identification of multiple forms of the noncapsid parvovirus protein NCVP1 in H-1 parvovirus-infected cells.

Analysis of extracts of H-1 parvovirus-infected cells with virus-specific antiserum led to the identification of two forms of the noncapsid virus protein NCVP1. These two proteins had apparent molecular weights of 84,000 (NCVP1) and 92,000 (NCVP1') and were structurally related, based on their immunological reactivity and on peptide map analysis. Both of these proteins appeared early in the virus infection, about the same time that capsid proteins appeared. NCVP1' was a highly phosphorylated protein which was apparently derived from NCVP1 via a post-translational event. Phosphoserine was the predominant phosphorylated amino acid in NCVP1' and appeared to be localized in one site or a few sites on the protein. The possible involvement of these noncapsid proteins in parvovirus DNA replication is discussed.

Amino Acid Sequence↗
PubMed1985

KBSH parvovirus: comparison with porcine parvovirus.

We compared the molecular, antigenic, and pathogenic properties of KBSH parvovirus to those of porcine parvovirus (PPV) isolate NADL-8. KBSH, propagated in swine testes cells in culture, possessed two major capsid polypeptides of 83 and 64 kilodaltons that were similar in size to those of PPV. KBSH-infected cells also contained an 86-kilodalton nonstructural polypeptide that was identical in size to the PPV nonstructural polypeptide (NS-1). The KBSH polypeptides were structurally similar but not identical to the corresponding PPV polypeptides, as revealed by partial proteolysis mapping. Viral replicative-form DNA from KBSH-infected cells was similar in size to PPV replicative-form DNA and exhibited similar but not identical restriction endonuclease cleavage patterns to that of PPV replicative-form DNA. Antigenically, the two viruses were also very closely related. By using heterologous and homologous antisera, the two viruses were indistinguishable in hemagglutination inhibition and immunoprecipitation assays. However, pathogenically these viruses were dramatically different. NADL-8 caused fetal death when injected into swine fetuses in utero and viremia and high persisting antibody titers when administered orally to weaning-age swine. KBSH-inoculated fetuses were normal in appearance, and pigs orally exposed to KBSH failed to establish viremia and demonstrated only transient antibody titers. Thus, KBSH appears to be a PPV that is very closely related to a highly pathogenic PPV isolate, yet is itself nonpathogenic in swine. This reduced pathogenic potential of KBSH may be attributable to its poor ability to replicate in swine.

Animals↗
PubMed1997

Two parvoviruses that cause different diseases in mink have different transcription patterns: transcription analysis of mink enteritis virus and Aleutian mink disease parvovirus in the same cell line.

The two parvoviruses of mink cause very different diseases. Mink enteritis virus (MEV) is associated with rapid, high-level viral replication and acute disease. In contrast, infection with Aleutian mink disease parvovirus (ADV) is associated with persistent, low-level viral replication and chronic severe immune dysregulation. In the present report, we have compared viral transcription in synchronized CRFK cells infected with either MEV or ADV using a nonradioactive RNase protection assay. The overall level of viral transcription was 20-fold higher in MEV- than in ADV-infected cells. Furthermore, MEV mRNA encoding structural proteins (MEV mRNA R3) was dominant throughout the infectious cycle, comprising approximately 80% of the total viral transcription products. In marked contrast, in ADV-infected cells, transcripts encoding nonstructural proteins (ADV mRNA R1 and R2) comprised more than 84% of the total transcripts at all times after infection, whereas ADV mRNA R3 comprised less than 16%. Thus, the ADV mRNA coding for structural proteins (ADV mRNA R3) was present at a level at least 100-fold lower than the corresponding MEV mRNA R3. These findings paralleled previous biochemical studies analyzing in vitro activities of the ADV and MEV promoters (J. Christensen, T. Storgaard, B. Viuff, B. Aasted, and S. Alexandersen, J. Virol. 67:1877-1886, 1993). The overall low levels of ADV mRNA and the paucity of the mRNA coding for ADV structural proteins may reflect an adaptation of the virus for low-level restricted infection.

Aleutian Mink Disease Virus↗