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PubMed1991

Detection of B19 parvovirus in human fetal tissues by electron microscopy.

We present the electron microscopy observations on samples from 38 pregnancies that were investigated for B19 parvovirus infection. Thirty-four had resulted in fetal loss thought to be due to a virus infection and 22 of the 38 were positive for B19 parvovirus in one or more of the tissues. Twenty-one placentas and 75 fetal tissue samples were examined. Fresh samples were investigated by immune electron microscopy while formalin-fixed tissues were examined as thin sections and by negative staining of tissue extracts with direct electron microscopy. Electron microscopy was more sensitive on fresh than on fixed samples. The ultrastructural observations on thin sections of fixed tissues yielded new information locating B19 parvovirus particles in both nucleus and cytoplasm of infected fetal cells. The diagnostic results of the range of electron microscopy assays were compared with those of two hybridization methods. The fresh samples yielded comparable results from electron microscopy and hybridisation assays but on formalin-fixed materials hybridisation was more sensitive.

Erythema Infectiosum↗

PubMed2006

Concurrent detection of human herpesvirus type 6 and measles-specific IgMs during acute exanthematic human parvovirus B19 infection.

A familial outbreak of human parvovirus B19 infection is described in which serological tests carried out routinely for determining the causal agent of febrile rashes of viral etiology failed to yield a definitive diagnosis. Concurrent detection of serum IgMs to parvovirus B19 and to heterologous viruses such as human herpesvirus type 6 (HHV-6) and measles virus complicated interpretation of the data. IgG avidity tests and investigation and testing for the presence of viral DNA in sera by PCR were required to confirm parvovirus B19. The study stresses the importance of avidity and PCR tests to obtain a firm diagnosis of febrile exanthematic viral diseases.

Antibodies, Viral↗

PubMed2005

Hydrops fetalis and neonatal death from human parvovirus B19: an unusual complication.

A case of prenatally diagnosed human parvovirus B19 (HPVB19) infection is reported. The neonate died after intrauterine therapy and premature delivery. The fetus was diagnosed with oedema, cardiomegaly, poor myocardial contractility and a pericardial effusion at 24/40 weeks' gestation. Ultrasound using colour flow Doppler showed a midcerebral artery peak systolic velocity (MCA PSV) raised at 45 cm/s, suggesting fetal anaemia. This was confirmed on fetal blood sampling, but recovery was suggested with a reticulocyte count of 16.8%. The fetal karyotype was normal, 46,XY. Fetal IgM was positive for Parvovirus. A week later, severe fetal anaemia was suspected and intrauterine transfusion carried out. Altogether three transfusions were given. At 31/40 weeks, the mother presented to her local hospital with suspected preterm labour, a caesarean section was carried out because of fetal compromise on cardiotocography. The baby was in poor condition at birth and resuscitation was stopped at 45 min of age. The post-mortem examination confirmed the hydrops and proved persistent Parvovirus infection, cardiac involvement and severe liver fibrosis.HPVB19 generally follows a benign course with intrauterine therapy; however, in this case, the fetus died despite successful transfusions. The reasons for this are discussed.

Adult↗

PubMed1995

Prevalence and clinical significance of anticardiolipin antibodies in pregnancies complicated by parvovirus B19 infection.

Anticardiolipin antibodies were measured in 60 pregnant women with acute parvovirus B19 infection. Test results for eight (13.3 per cent) women were positive for anticardiolipin antibody. Six of these eight women became negative later, yielding a prevalence of anticardiolipin antibodies of 3.3 per cent (2/60) 6 months after acute parvovirus B19 infection. Anticardiolipin antibody positivity was not associated with an increased risk of abortion, fetal death, or maternal complications. This study suggests that there is an elevated frequency of anticardiolipin antibodies in pregnant women with acute parvovirus B19, probably representing an epiphenomenon. However, this is not associated with an adverse maternal or perinatal outcome.

Antibodies, Anticardiolipin↗

PubMed1996

Parvoviruses and bone marrow failure.

Parvovirus B19, the only known human pathogenic parvovirus, is highly tropic to human bone marrow and replicates only in erythroid progenitor cells. The basis of this erythroid tropism is the tissue distribution of the B19 cellular receptor, globoside (blood group P antigen). In individuals with underlying hemolytic disorders, infection with parvovirus B19 is the primary cause of transient aplastic crisis. In immunocompromised patients, persistent B19 infection may develop that manifests as pure red cell aplasia and chronic anemia. B19 infection in utero can result in fetal death, hydrops fetalis or congenital anemia. Diagnosis is based on examination of the bone marrow and B19 virological studies. Treatment of persistent infection with immunoglobulin leads to a rapid, marked resolution of the anemia.

Adult↗

PubMed1997

The removal of model viruses, poliovirus type 1 and canine parvovirus, during the purification of human albumin using ion-exchange chromatographic procedures.

The manufacturing process for albumin in Australia is based primarily on ion-exchange chromatography. The capacity of ion-exchange matrices to remove non-enveloped viruses (canine parvovirus and poliovirus type 1) was assessed using a scaled-down chromatographic process which was shown to yield product meeting purity criteria set for the manufacturing process. Poliovirus type 1 and canine parvovirus were added at one tenth the volume of desalted and delipidated Supernatant II + III produced by traditional Cohn Fractionation from human plasma before the material was applied to DEAE and CM ion-exchangers connected in series. Samples were taken at equilibration, wash, elution and regeneration steps and the log clearance and reduction of the viruses calculated. The mean clearance and reduction factors for viral load of poliovirus type 1 were 5.3 logs and 3.2 logs, respectively and 1.8 logs and 1.8 logs for canine parvovirus.

Animals↗

PubMed2000

Expression of the insect parvovirus GmDNV in vivo: the structural and nonstructural proteins are encoded by opposite DNA strands.

The nucleotide sequence of GmDNV, an insect parvovirus, reveals large open reading frames (ORFs) on both strands of the viral replicative form DNA. Previously, we identified two viral transcripts within the polyadenylated RNA fraction of infected host larvae (Gross et al., 1990, J. Invertebr. Pathol. 56, 175-180). In this work we used hybridization of single-stranded, unidirectional probes to RNA blots to show that the two transcripts, synthesized in vivo in GmDNV-infected Galleria mellonella larvae, are of antiparallel orientation. To determine their coding specificities, polyadenylated RNAs were isolated from hybrids with DNA from the left and right halves of the viral genome and translated in a rabbit reticulocyte system. The "right," 2.4-kb hybrid-selected RNA was shown to direct the synthesis of four polypeptides that comigrated with the four viral capsid proteins and were immunoprecipitated with anti-GmDNV serum. Translation of the "left," 1.8-kb RNA yielded three polypeptides, none of which was detected among the viral capsid proteins. This type of expression strategy is unique among vertebrate and most invertebrate parvoviruses, which use only one DNA strand to encode all their proteins. On the other hand, the basic organization of parvoviruses, in which the regulatory and structural proteins are encoded, respectively, by two clusters of ORFs located at the left and right halves of the genome, is conserved.

Animals↗

PubMed1996

Evolution of canine parvovirus involved loss and gain of feline host range.

Canine parvovirus (CPV) type-2 emerged as a new virus infecting dogs in 1978, and it was probably derived as a variant of feline panleukopenia virus or of a closely related virus infecting another carnivore. CPV type-2 was subsequently replaced in nature by antigenically variant viruses (CPV type-2a and CPV type-2b) which now coexist in dog populations worldwide. We show that CPV type-2 isolates did not replicate in cats, but that both CPV type-2a and CPV type-2b isolates replicated efficiently. About 10% of the viruses isolated from cats with natural parvovirus disease were antigenically indistinguishable from CPV type-2a or type-2b. The capsid protein gene sequence of a 1990 feline parvovirus isolate ("FPV-24") was essentially identical to the sequence of CPV type-2b viruses from dogs. The loss and reacquisition of the feline host range in CPV was most likely due in each case to small numbers of changes in a region of the virus capsid where three protein monomers interact.

Animals↗

PubMed1998

Nonstructural protein-2 and the replication of canine parvovirus.

The nonstructural protein-2 (NS2) of canine parvovirus (CPV) is produced from the left-hand open reading frame of the viral genome and contains 87 amino-terminal amino acids in common with nonstructural protein 1 (NS1) joined to 78 amino acids from an alternative open reading frame. In the minute virus of mice parvovirus NS2 plays a role in controlling capsid protein assembly and translation in a host-specific manner. The predicted NS2 of CPV is divergent from the proteins of the rodent parvoviruses, and the protein and its functions have not been described. We characterized the large and the small splices of CPV using reverse transcriptase-PCR, NS2 was identified using anti-peptide antibodies against the predicted C-terminal sequence and also by expressing the protein from a plasmid vector. The protein could be detected at low levels in the nucleus and the cytoplasm of a proportion of CPV-infected cells, as well as in cells transfected with the expression plasmid. Virus genomes were prepared with mutations in the splice donor or acceptor sites of the NS2-specific intron or with three different termination codons in the NS2-unique exon. Both splice donor and acceptor mutations resulted in the use of previously cryptic splice sites, and the virus containing the splice donor mutation replicated inefficiently. However, the other four mutant viruses were all viable and replicated efficiently in cat and dog cells, and two mutant viruses that were tested appeared to assemble their capsids in the same manner as did the wildtype. After inoculation of dogs an NS2 mutant virus with a termination codon in the NS2-unique exon replicated to titers similar to those seen for wildtype CPV in several tissues examined.

Alternative Splicing↗

PubMed2000

Rhesus and pig-tailed macaque parvoviruses: identification of two new members of the erythrovirus genus in monkeys.

We have previously reported the identification of a novel simian parvovirus in cynomolgus monkeys, which causes severe anemia in immunosuppressed cynomolgus monkeys and is currently being studied as an animal model for human B19 infection. We now report two similar outbreaks of anemia in rhesus and pig-tailed macaques associated with two distinct but similar simian parvoviruses (pig-tailed macaque and rhesus parvovirus). Both viruses have been cloned and over 5000 nucleotides sequenced from each virus. The viruses show marked similarities to other members of the Erythrovirus genus in the Parvoviridae family.

Amino Acid Sequence↗

PubMed2000

Predominance of canine parvovirus (CPV) in unvaccinated cat populations and emergence of new antigenic types of CPVs in cats.

Serological, sequence, and in vitro host range analyses of feline parvovirus (FPV) isolates in Vietnam and Taiwan revealed that more than 80% of the isolates were of the canine parvovirus (CPV) type, rather than feline panleukopenia virus (FPLV). Although parvovirus isolates from three Vietnamese leopard cats were genetically related to CPV type 2a or 2b, they had a natural mutation of VP2 residue 300 Gly to an Asp, resulting in remarkable changes in their antigenic properties. These results indicated the possibility that CPV-2a/2b-type viruses can spread in cats more efficiently than conventional FPLV under natural conditions and that CPV-2a/2b viruses are further evolving in cats.

Amino Acid Sequence↗

PubMed2006

Parvovirus B19: the causative agent of dilated cardiomyopathy or a harmless passenger of the human myocard?

Parvovirus B19 infections may cause a widespread benign and self-limiting disease in children and adults known as erythema infectiosum (fifth disease). Several further manifestations are associated with B19 infections, such as arthralgias, arthritis, leucopenia and thrombocytopenia, anaemia and vasculitis and spontaneous abortion and hydrops fetalis in pregnant women. Persistent infections with continuous virus production may occur in immunocompetent as well as in immunosuppressed individuals. Parvovirus B19 infections have been frequently implicated as a cause or trigger of various forms of autoimmune diseases affecting joints, connective tissue and large and small vessels. Autoimmune neutropenia, thrombocytopenia and haemolytic anaemia are known as sequelae of B19 infections. The molecular basis of the autoimmune phenomena is unclear. Many patients with these long-lasting symptoms are not capable of eliminating the virus or controlling its propagation. Furthermore, latent viral genomes have been detected in cells of various organs and tissues by PCR. At present, it is not clear if these cells produce viral proteins and/or infectious B19 particles, if the virus genome can be reactivated to productive replication and if the presence of viral DNA indicates a causative role of parvovirus B19 with distinct diseases.

Animals↗

PubMed2006

Oncolytic murine autonomous parvovirus, a candidate vector for glioma gene therapy, is innocuous to normal and immunocompetent mouse glial cells.

The sensitivity of brain tumour cells to wild-type or recombinant parvoviruses H1-PV and MVMp makes these agents promising candidates for gene therapy of astrocytoma. This application raises the question of whether parvoviruses exert deleterious or bystander effects on normal glial cells surrounding tumours. We addressed this question in the mouse model by using cell cultures derived from BALB/c, C57BL/6 and VM/Dk strains. Astrocytes and a large proportion of microglia cultures were competent for MVMp uptake. Infection was, however, abortive as replication-associated viral proteins synthesis took place in less than 10% of astrocytes and no progeny virions were produced. This restriction was even more pronounced for microglia in which no viral protein expression could be detected, save for a minute fraction of VM/Dk-derived cells. Infection with MVMp had no significant effect on glial cell survival and did not interfere with their immune potential. Indeed, neither the lipopolysaccharide (LPS)/interferon (IFN-gamma)-induced cytotoxicity of VM/Dk-derived microglia towards the mouse glioma (MT539MG) cell line, nor the glial cells capacity for tumour necrosis factor alpha production upon LPS stimulation or LPS/IFN-gamma stimulation were affected by infection with MVMp. Moreover, stimulation with LPS and/or IFN-gamma resulted in a decreased expression of the viral replicative and cytotoxic protein NS1. Together, our data indicate that, in the natural host, a majority of normal glial cells are not competent for MVMp replication and that the abortive infection taking place in a minor fraction of these cells fails to impede their survival and immunocompetence, giving credit to the consideration of autonomous parvoviruses for glioma therapy.

Animals↗

PubMed1997

Chronic anemia as a complication of parvovirus B19 infection in a pediatric kidney transplant patient.

This is a report of unexplained anemia that persisted for 4 months in an adolescent renal transplant patient receiving immunosuppression that included prednisone, tacrolimus, and mycophenolate mofetil. This patient required monthly blood transfusions for fatigue, palpitations, and hematocrit levels between 15% and 17%. In addition, his posttransplant course was notable for the development of insulin-dependent diabetes mellitus. While receiving low-dose prednisone, he was switched from tacrolimus to cyclosporin and tapered off insulin injections over the next 2 months. At 4.5 months post-transplantation, further diagnostic evaluation was suggestive of parvovirus B19 infection as the cause for our patient's chronic anemia. After testing negative for serum-specific parvovirus B19 IgM and IgG antibodies, parvovirus B19 infection was detected in blood by the polymerase chain reaction. Treatment with intravenous immunoglobulin (1 g/kg per day x 2 days) resulted in normalization of both his reticulocyte count and hematocrit within 6 weeks. At 4 months after receiving the immunoglobulin infusion, he has maintained a normal hematocrit level and stable renal function without requiring further blood transfusions.

Adolescent↗

PubMed1999

Parvovirus glomerulonephritis in a patient with sickle cell disease.

A child with sickle cell disease developed glomerulonephritis 10 days following an aplastic crisis induced by human parvovirus B 19 infection. An initial kidney biopsy showed focal proliferative glomerulonephritis, and 1 year later was compatible with focal and segmental glomerulosclerosis. Renal tissue, tested by polymerase chain reaction, was positive for parvovirus, while the patient's blood was negative. For the first time a direct relationship has been demonstrated between parvovirus infection and glomerulonephritis.

Adolescent↗

PubMed2000

High incidence of parvovirus B19 DNA in synovial tissue of patients with undifferentiated mono- and oligoarthritis.

A common problem in rheumatological practice is inflammatory joint disease that cannot be classified. The prognosis of such undifferentiated arthritides is uncertain. The synovial tissue of 41 consecutive patients with various forms of arthritis was tested for the presence of viral DNA in a diagnosis-unaware fashion, using the polymerase chain reaction (PCR). Of all tested viruses, cytomegalovirus and parvovirus B19 were positive (each in 10 patients, two double-positives), whereas herpes simplex virus was positive in two patients. Rubella virus RNA was detected in three specimens. When the positivity for viral material was analysed in terms of distribution among the various diagnostic groups, it became evident that five out of 10 parvovirus B 19-positive patients belonged to the undifferentiated arthritis group, whereas cytomegalovirus-positive patients were spread among all diagnostic groups. This indicates the possibility of a new diagnostic category of undifferentiated mono- and oligoarthritis, which can be identified by the presence of parvovirus B19 DNA in synovial tissue.

Adolescent↗

PubMed1995

Detection of human parvovirus B19 DNA by nested polymerase chain reaction.

We describe the development of a nested polymerase chain reaction (PCR) technique used to detect human parvovirus B19 DNA. It was performed in a two-step reaction, first with a pair of outer primers, then with a pair of inner (ie, nested) primers. Primer sequences were selected in the VP1 gene, corresponding to the capsid protein, of human parvovirus B19. To establish the nested PCR assay, the plasmid pGEM-1 containing almost the entire coding sequence of a human parvovirus B19 isolate was used. The nested PCR could detect up to 1.8 x 10(-3) ag of B19 DNA, equivalent to 10(-4) genomes, by electrophoresis. No amplification product was detected by gel electrophoresis when the reaction mixture contained human placental DNA, cytomegalovirus DNA, and sterile distilled water as templates. We used this assay to evaluate four cases of maternal B19 infection that were diagnosed by determination of the presence of anti-B19 immunoglobulin-M in maternal serum. The advantages of our nested PCR for detecting B19 DNA are plating simplicity, safety, sensitivity, and specificity. Our results suggest that this method may have general applicability in the evaluation of nonimmune hydrops fetalis and in the documentation of the natural history of fetal infection with B19 when applied to specimens of amniotic fluid or fetal blood.

Adult↗

PubMed1995

A simple touch-down polymerase chain reaction for the detection of canine parvovirus and feline panleukopenia virus in feces.

A polymerase chain reaction (PCR) assay is described for the detection of parvovirus in feces of dogs and cats. A touch-down protocol was used which enabled the specific amplification of virion DNA from feces after a fast and simple boiling pretreatment. The sensitivity of PCR was as high as ten infectious particles per reaction which corresponds to a titer of about 10(3) infectious particles per gram of unprocessed feces. This renders the PCR about 10- to 100-fold more sensitive than electron microscopy, the standard method for parvovirus diagnosis. The very rapid and simple sample preparation recommends this PCR assay as an alternative technique for routine parvovirus diagnosis.

Animals↗