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PubMed1996

Efficient induction of persistent and prenatal parvovirus infection in rats.

Parvoviruses are prevalent and disruptive infectious agents of laboratory rats. Risks to rat-based research from infection are increased by the persistence of virus in immune rats and by prenatal transmission of infection. The mechanisms leading to viral persistence and prenatal infection are poorly understood and have been difficult to study for lack of reliable and humane induction methods. We report here protocols for inducing persistent and prenatal infection without causing clinical disease using the UMass strain of rat virus (RV), a common rat parvovirus. Infant rats inoculated by the oronasal route at 6 days of age had greater than 90% prevalence of persistent infection. RV-UMass also induced intrauterine infection in pregnant rats inoculated by the oronasal route. Inoculation of dams at gestation day 9 frequently caused severe disease in the fetuses whereas inoculation at gestation day 12 caused primarily asymptomatic fetal infection that persisted post partum RV-UMass infection facilitates study of parvoviralhost interactions that are relevant to laboratory rats and which also may improve understanding of persistent and prenatal human parvovirus infection.

Animals↗

PubMed1995

Parvovirus B19 infection and hematopoiesis.

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 (TAC). 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.

Animals↗

PubMed1993

Antigenic and genomic variabilities among recently prevalent parvoviruses of canine and feline origin in Japan.

Canine parvovirus type 2 (CPV-2) and feline panleukopenia (FLP) virus (FPLV) are well known and ubiquitous diarrhea-causing pantropic viruses. A "new" antigenic variant of CPV-2 (designated as CPV-2a) has been also prevalent among dogs in Japan. In the present study, 24 canine and 8 feline isolates collected during 1987-1991 were compared with 17 CPV-2 or CPV-2a and 7 FPLV strains that had been characterized previously. Genomic properties were determined by the restriction cleavage patterns of amplified genes encoding the capsid proteins VP1 and VP2 by the polymerase chain reaction. Antigenic properties were determined by hemagglutination-inhibition assay with monoclonal antibodies against an FPLV strain. Growth characteristics in feline CRFK and canine MDCK cells were also examined. Genomic and antigenic properties of the canine isolates were relatively invariable with one exceptional isolate, C27, which was recovered from a typical clinical case of parvovirus infection but possessed properties similar to FPLV rather than CPV-2 and CPV-2a. All isolates from FPL cases possessed the same genomic and antigenic properties as those of reference FPLVs isolated in the 1970s, but three of five strains isolated from the feces of clinically healthy cats were likely to be of canine origin because they possessed very similar properties to CPV-2a. Although species-specificity of these novel isolates could not be determined definitely, the results indicate a possibility that transmission of parvovirus has occurred between these two animal species.

Animals↗

PubMed2005

Molecular and structural characterization of fluorescent human parvovirus B19 virus-like particles.

Although sharing a T=1 icosahedral symmetry with other members of the Parvoviridae family, it has been suggested that the fivefold channel of the human parvovirus B19 VP2 capsids is closed at its outside end. To investigate the possibility of placing a relatively large protein moiety at this site of B19, fluorescent virus-like particles (fVLPs) of B19 were developed. The enhanced green fluorescent protein (EGFP) was inserted at the N-terminus of the structural protein VP2 and assembly of fVLPs from this fusion protein was obtained. Electron microscopy revealed that these fluorescent protein complexes were very similar in size when compared to wild-type B19 virus. Further, fluorescence correlation spectroscopy showed that an average of nine EGFP domains were associated with these virus-like structures. Atomic force microscopy and immunoprecipitation studies showed that EGFP was displayed on the surface of these fVLPs. Confocal imaging indicated that these chimeric complexes were targeted to late endosomes when expressed in insect cells. The fVLPs were able to efficiently enter cancer cells and traffic to the nucleus via the microtubulus network. Finally, immunoglobulins present in human parvovirus B19 acute and past-immunity serum samples were able to detect antigenic epitopes present in these fVLPs. In summary, we have developed fluorescent virus-like nanoparticles displaying a large heterologous entity that should be of help to elucidate the mechanisms of infection and pathogenesis of human parvovirus B19. In addition, these B19 nanoparticles serve as a model in the development of targetable vehicles designed for delivery of biomolecules.

Animals↗

PubMed2004

Report of a collaborative study to calibrate the Second International Standard for parvovirus B19 antibody.

A collaborative study was undertaken to assess the suitability of a replacement for the First International Standard for parvovirus B19 IgG, human serum and to calibrate it in IU. The proposed standard, which is a pool of sera from 16 US blood donors, was assayed along with the First International Standard, a coded duplicate of the proposed standard and a plasma sample from a single blood donor. Nine laboratories from eight countries participated in the studies and five different assay kits were used. Two kits contained VP1+VP2, one kit contained VP1 only and two kits, one of which was used by five participants contained VP2 only. Differences in detection of the proposed standard and the individual plasma were observed with assay kits containing different antigens, VP1, VP2 or VP1+VP2. However, since VP1 is a minor capsid protein and on its own does not assemble into virus like particles and the dominant response in individuals appears to be against VP2, it was considered reasonable to utilize only the data from kits containing VP2 antigen for the calibration of the proposed standard. The results of this study demonstrated that the proposed standard coded 01/602 was suitable to serve as the replacement International Standard for parvovirus B19, serum IgG, and this preparation was established as the Second International Standard for parvovirus B19 antibody, plasma human, with an assigned unitage of 77 IU per ampoule by the Expert Committee on Biological Standardisation of the World Health Organisation in February 2003.

Calibration↗

PubMed2004

Replicative multivirus infection with cytomegalovirus, herpes simplex virus 1, and parvovirus B19, and latent Epstein-Barr virus infection in the synovial tissue of a psoriatic arthritis patient.

BACKGROUND: Psoriatic arthropathy occurs as complicating feature in about 5-7% of psoriasis patients. Infectious mechanisms including viral antigens have been suggested by serologic data as CD8 T cellular specifity towards viral epitopes. OBJECTIVE AND RESULTS: We here reported a case of a 32-year-old male psoriatic arthritis patient, where we could demonstrate simultaneous infection with cytomegalovirus (CMV), herpes simplex virus type I (HSV1) and parvovirus B19 (B19), as well as latent Epstein-Barr virus (EBV) infection within the synovial tissue by immunohistochemistry (CMV, parvovirus B19, HSV1, EBV-LMP) and DNA-in situ-hybridization (CMV). Serologic examination revealed positive EBV and parvovirus B19-IgG-antibodies, but no antibody response to HSV1 and CMV. CONCLUSION: This case is of special interest, since replicative viral infections have not yet been demonstrated localised in the psoriatic arthritis synovia. Thus, with particular regard to the limited information of the serologic data and the possible need of immuno suppressive therapy direct synovial testing for viral antigenes may be considered in psoriatic arthritis patients.

Adult↗

PubMed2004

Prevalence of human parvovirus B19 DNA in cardiac tissues of patients with congenital heart diseases indicated by nested PCR and in situ hybridization.

BACKGROUND: Infection with parvovirus B19 (B19) was reported to be correlated with myocarditis, cardiomyopathy, and kawasaki disease. But no information is available about the relationship between inutero B19 infection and congenital heart disease (CHD). OBJECTIVE: To explore whether there is relationship between B19 infection and CHD. STUDY DESIGN: Retrospective investigation of biopsy samples from CHD patients from January 1996 to December 1998. METHODS: Parvovirus B19 was detected in biopsy samples from 42 cases of CHD patients and 38 cases of biopsy or autopsy samples from patients with other diseases (controls) by nested polymerase chain reaction (PCR) and in situ hybridization (ISH) technique. HE staining was also performed to observe the morphology of these cardiac tissue samples. RESULTS: Nested PCR assay indicated that seven of 42 (16.7%) CHD cardiac tissue were B19 DNA positive, while all the 38 controls were B19 DNA negative, the difference is significant (P = 0.012). ISH assay indicated that five of 42 (11.7%) CHD cardiac tissues were positive for B19 DNA and none of the control cardiac tissue were positive, all B19 DNA positive signals were located in the nucleuses of cardiac cells. HE staining showed that there was no inflammatory change in B19 DNA positive cardiac tissue. CONCLUSIONS: Parvovirus B19 DNA was presented in part of CHD cardiac tissues and located in nucleus, which suggested that inutero B19 infection might be correlated with CHD.

Adolescent↗

PubMed2005

High frequency of parvovirus B19 DNA in bone marrow samples from rheumatic patients.

BACKGROUND: Human parvovirus B19 (B19) polymerase chain reaction (PCR) is now a routine analysis and serves as a diagnostic marker as well as a complement or alternative to B19 serology. The clinical significance of a positive B19 DNA finding is however dependent on the type of tissue or body fluid analysed and of the immune status of the patient. OBJECTIVES: To analyse the clinical significance of B19 DNA positivity in bone marrow samples from rheumatic patients. STUDY DESIGN: Parvovirus B19 DNA was analysed in paired bone marrow and serum samples by nested PCR technique. Serum was also analysed for B19-specific IgG and IgM antibodies and the results were compared with clinical and epidemiological data. RESULTS AND CONCLUSIONS: B19 IgG was found in 41 of 50 patients (82%) whereas none was B19 IgM positive. The serologic evaluation showed that none of the patients had acute B19 infection. However, B19 DNA was detected by PCR in 13 of 50 (26%) bone marrow samples from these patients indicating a high frequency of persistent infection compared with previous reports of patient groups and healthy controls. In the study, 22 patients had rheumatoid arthritis (RA) and 7 of these RA patients were B19 DNA positive in bone marrow. Rheumatoid factor was positive in 4 of the 7 B19 DNA positive RA patients as compared with Rheumatoid factor positivity in all of the 15 B19 DNA negative RA patients. Erosive arthritis in X-ray was less common in the B19 DNA positive group than in the B19 DNA negative group. A high frequency of parvovirus B19 DNA was thus detected in bone marrow samples in rheumatic patients. The clinical data does not support a direct association between B19 PCR positivity and rheumatic disease manifestation. Therefore, the clinical significance of B19 DNA positivity in bone marrow samples from rheumatic patients must be interpreted with caution.

Adolescent↗

PubMed2005

Parvovirus B19 viral loads in relation to VP1 and VP2 antibody responses in diagnostic blood samples.

BACKGROUND: Human parvovirus B19 infection is characterised by high peak viral load levels followed by episodes of prolonged viremia. The risk of transmission of parvovirus B19 by blood or blood products has been increasingly recognised and parameters that can predict the risk of transmission are subject of interest. OBJECTIVES: This study aimed to study correlations between B19 viral DNA loads and antibody responses to the viral antigens VP1 and VP2 in clinical serum samples. STUDY DESIGN: A panel of 1610 serum samples from patients clinically suspected from acute B19 infection were analysed. Antibodies were measured by the parvovirus anti-VP1 immuno-fluorescence assay (IFA) and the anti-VP2 enzyme immunoassay (EIA) from Biotrin. B19 viral loads were measured by a real-time PCR using the external WHO standard for DNA quantification. RESULTS: Positive IgM responses were found in 154 (9.6%) of the 1610 sera tested. Based on the PCR results in a subset of 312 sera, the anti-VP2 EIA IgM showed a better combination of sensitivity/specificity (91%/94%) compared to the anti-VP1 IFA (66%/97%). B19 DNA levels in the sera strongly correlated with the levels of IgM antibodies, all sera with high viral loads (>10(6)IU/ml) having VP2 EIA IgM ratios above 3.0. CONCLUSIONS: The B19 VP2 IgM ELISA is superior to the B19 VP1 IgM IFA if verified by PCR. Anti-VP2 IgM antibodies in sera are indicative for the presence B19 DNA and can be used to predict high levels of B19 DNA in diagnostic sera.

Adolescent↗

PubMed2006

Genetic variants of parvovirus B19 identified in the United Kingdom: implications for diagnostic testing.

BACKGROUND: Discrepant results in diagnostic parvovirus B19 PCR assays have been observed with strains showing nucleotide sequence variation. OBJECTIVES AND STUDY DESIGN: To perform phylogenetic analysis on two parvovirus B19 strains that gave discrepant PCR results. RESULTS: One strain was found to be genotype 2; the second strain was genotype 3. CONCLUSIONS: Parvovirus B19 genotypes 2 and 3 strains were identified in diagnostic samples of UK origin following the investigation of discrepant PCR results. More structured investigations are needed to estimate the prevalence of these variants. In the meantime, diagnostic PCR results should be interpreted cautiously when they are at variance with serological testing. Manufacturers of PCR kits for the detection of B19 sequences will need to consider re-designing their primers.

Adult↗

PubMed2005

Parvovirus B19 infection in an isolated pancreas transplant recipient.

We report an isolated pancreas transplant recipient on immunosuppressive therapy with parvovirus B19 infection. He presented with a chronic transfusion-dependent anemia, unresponsive to erythropoietin therapy. Bone marrow cytomorphology was highly suggestive of parvovirus pure red cell aplasia, which was confirmed with polymerase chain reaction positive for parvovirus B19 DNA from peripheral blood. The anemia responded briskly and reticulocyte counts improved from 0.0% to 17.0% within 1 week after the administration of intravenous immunoglobulin. We discuss the difficulty of serological diagnosis in such cases, the importance of using techniques that directly identify the virus, and taking measures that may prevent recurrence.

Adult↗

PubMed2006

Occurrence of severe gastroenteritis in pups after canine parvovirus vaccine administration: a clinical and laboratory diagnostic dilemma.

A total of 29 faecal samples collected from dogs with diarrhoea following canine parvovirus (CPV) vaccination were tested by minor groove binder (MGB) probe assays for discrimination between CPV vaccine and field strains and by diagnostic tests for detection of other canine pathogens. Fifteen samples tested positive only for CPV field strains; however, both vaccine and field strains were detected in three samples. Eleven samples were found to contain only the vaccine strain, although eight of them tested positive for other pathogens of dogs. Only three samples were found to contain the vaccine strain without evidence of canine pathogens. The present study confirms that most cases of parvovirus-like disease occurring shortly after vaccination are related to infection with field strains of canine parvovirus type 2 (CPV-2) rather than to reversion to virulence of the modified live virus contained in the vaccine.

Animals↗

PubMed2005

Parvovirus B19 does not bind to membrane-associated globoside in vitro.

The glycosphingolipid globoside (globotetraosylceramide, Gb4Cer) has been proposed to be the cellular receptor of human parvovirus B19. Quantitative measurements of the binding of parvovirus B19 to Gb4Cer were performed to explore the molecular basis of the virus tropism. Solid-phase assays with fluorescence-labeled liposomes or 125iodine-labeled empty capsids were used to characterize the specificity of binding. In addition, surface plasmon resonance on lipid layers, as well as isothermal titration microcalorimetry, was utilized for real-time analysis of the virus-receptor interaction. These studies did not confirm binding of Gb4Cer to recombinant B19 VP2 capsids, suggesting that Gb4Cer does not function on its own as the cellular receptor of human parvovirus B19, but might be involved in a more complex recognition event. The biochemical results were further confirmed by cryo-electron microscopy image reconstructions at 10 A resolution, in which the structures of empty capsids were compared with empty capsids incubated with Gb4Cer.

Animals↗

PubMed2005

Adeno-associated virus type 2 enhances goose parvovirus replication in embryonated goose eggs.

The autonomous goose parvovirus (GPV) and the human helper-dependent adeno-associated virus type 2 (AAV2) share a high degree of homology. To determine if this evolutionary relationship has a biological impact, we studied viral replication in human 293 cells and in embryonated goose eggs coinfected with both viruses. Similar experiments were performed with the minute virus of mice (MVM), an autonomous murine parvovirus with less homology to AAV2. In human 293 cells, both GPV and MVM augmented AAV2 replication. In contrast, AAV2 markedly enhanced GPV replication in embryonated goose eggs under conditions where a similar effect was not observed with MVM. AAV2 did not replicate in embryonated goose eggs and AAV2 inactivated by UV-irradiation also enhanced GPV replication. To our knowledge, this is the first report that a human helper-dependent member of the Parvoviridae can provide helper activity for an autonomous parvovirus in a natural host.

Animals↗

PubMed1998

Long-term outcome of children following maternal human parvovirus B19 infection.

OBJECTIVE: To determine the long-term outcomes of children exposed in utero to maternal parvovirus B19 infection. METHODS: All pregnant women with serologic evidence of recent parvovirus B19 infection and a comparison group with serologic evidence of past infection from January 1988 to December 1994 were sent questionnaires or contacted by phone about the health and development of their children. Information requested included: pregnancy complications, date of delivery, birth weight, sex, birth defects, need for special care, significant health problems, and developmental delays. All women had serology done at either the Centers for Disease Control and Prevention or the virology laboratory of the Connecticut Department of Health. The data were analyzed using descriptive statistics, chi2 analysis with Fisher exact test, or Student t test in appropriate cases. P < .05 was considered significant. RESULTS: Outcome information was obtained from 113 of 117 immunoglobulin-M positive women. The 113 respondents had 103 term singletons, two sets of twins (of which one neonate died of complications of prematurity), one hydropic stillborn, four spontaneous abortions, and one ectopic pregnancy. The mean gestational age at time of exposure was 15.6 weeks. The median age of the liveborn infants in study and comparison groups was 4 years. Eight of the 108 (7.3%) surviving children, one set of twins (exposed at 27 weeks), and six singletons (exposed at 7, 8, 9, 20, 27, and 35 weeks) had developmental delays in speech, language, information processing, and attention. Outcomes were obtained for 99 of 110 patients with past infection; they had 83 liveborn singletons, five sets of twins, two stillborns, and five spontaneous abortions. Seven of the 93 (7.5%) children had developmental delays, similar to the study group. Post-hoc power analysis revealed that 712 infected patients would be needed to find a twofold difference in the risk of abnormal neurologic development; our study had 30% power to find such a difference. CONCLUSION: There is no apparent increase in the frequency of developmental delays in children with exposure in utero to parvovirus, but larger studies are needed.

Adult↗

PubMed2001

Haematological response to parvovirus B19 infection in homozygous sickle-cell disease.

Infection with human parvovirus B19 is known to cause aplastic crises in patients with homozygous sickle-cell disease. We studied the haematological consequences of parvovirus B19 infection in 280 such patients who had been followed up from birth in Jamaica. Evidence of seroconversion was routinely sought with a baculovirus-based, enzyme immunoassay in serum samples taken during aplastic crises and in all stored annual serum samples. 70% of patients had seroconverted by age 20 years; of 177 infections, haematological change was typical of aplastic crises in 118 (67%), minor in 16 (9%), and not discernible in 43 (24%). This assay increased the detection of unsuspected seroconversion-an observation important in planning a strategy for parvovirus B19 immunisation.

Adolescent↗

PubMed1994

Chronic parvovirus B19 infection and systemic necrotising vasculitis: opportunistic infection or aetiological agent?

We describe three patients who had infection with human parvovirus B19 in association with new-onset systemic necrotising vasculitis syndromes, two with features of polyarteritis nodosa and one with features of Wegener's granulomatosis. Chronic B19 infection, lasting 5 months to more than 3 years, was shown by enzyme immunoassay for IgG and IgM antibodies to B19 and polymerase chain reaction for B19 DNA in serum and tissue samples. The patients had atypical serological responses to the B19 infection, although none had a recognisable immunodeficiency disorder. Treatment with corticosteroids and cyclophosphamide did not control vasculitis. Intravenous immunoglobulin (IVIG) therapy led to rapid improvement of the systemic vascultis manifestations, clearing of the chronic parvovirus infection, and long-term remission. These observations suggest an aetiological relation between parvovirus B19 infection and systemic necrotising vasculitis in these patients and indicate a potentially curative role for IVIG in such disorders.

Adolescent↗

PubMed1997

Persistence of parvovirus B19 DNA in synovial membranes of young patients with and without chronic arthropathy.

BACKGROUND: Human parvovirus B19 replicates in erythroid precursors of the bone marrow, and several diseases have been attributed to this virus including some cases of juvenile chronic arthropathy. METHODS: Tissue samples from children with juvenile arthritis and from healthy young adults with recent joint trauma were examined for B19 DNA by PCR. We also studied the timing of the parvovirus infection serologically. FINDINGS: All samples of synovial fluid, bone marrow, and blood were negative for B19 DNA. Eight (28%) of the 29 children with chronic arthritis had B19 DNA in synovial tissues. However, an even higher proportion of the non-arthropathy controls were positive for B19 DNA in synovial membranes (13 [48%] of 27). All the individuals with B19 DNA in synovial membrane had serum IgG antibodies to B19. INTERPRETATION: Genomic B19 DNA can persist in the synovial membranes not only in patients with chronic arthropathy but also in healthy immunocompetent individuals. The diagnostic criteria for parvovirus arthropathy must be reevaluated.

Adolescent↗