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PubMed2002

Molecular characterization of three newly recognized rat parvoviruses.

Rodent parvoviruses have been documented to interfere with both in vivo and in vitro research. In this study, three rat parvoviruses distinct from previously characterized rodent parvoviruses were identified from naturally infected rats obtained from four discrete sources. These three newly recognized parvoviruses were designated rat minute virus (RMV)-1a, -1b and -1c. In this study, the genomic nucleotide sequence and the predicted amino acid sequences of proteins for each of the three RMV-1 variants and Kilham rat virus (KRV) were determined and compared with previously characterized rodent parvoviruses. The three RMV-1 variants were shown to be closely related to each other, to be distinct from but closely related to KRV and H-1 virus, and to be significantly different from the previously identified rat parvovirus isolate, RPV-1a.

Amino Acid Sequence↗

PubMed2006

Short regions of sequence identity between the genomes of human and rodent parvoviruses and their respective hosts occur within host genes for the cytoskeleton, cell adhesion and Wnt signalling.

Our understanding of the mechanism(s) of pathogenesis and persistence of vertebrate parvoviruses remains incomplete. With the recent availability of the complete genome sequences of human, rat and mouse, and the ability to search these sequences and to locate matches to exact genomic regions, further insight into the interaction of parvoviruses with their human and rodent hosts is possible. To determine the extent and nature of sequence identity between candidate parvoviruses and their respective hosts, blast searches of the genome sequences of adeno-associated virus, parvovirus B19, mouse parvovirus, the prototype strain and immunosuppressant variant of minute virus of mouse, Kilham rat virus and rat parvovirus were performed against the genome(s) of their respective hosts (human, rat and mouse) using the resources of the NCBI and the Celera Discovery System. Regions of identity and similarity were mapped to their precise location in their particular host genome. For each virus, between one and 12 identical regions were found. Each identical region was 17-26 nt and was generally found at multiple sites within the particular host genome. These identical regions were predominantly located in non-coding regions of particular host genes and in intergenic regions. The ontology of host genes in which identical regions were found for each of the nine virus-host interactions highlighted several pathways/processes, including the cytoskeleton, cell adhesion and Wnt signalling. Within each virus species, these homologous regions were highly conserved (100 % identity in 16 out of 23 alignments where more than one sequence was available). All of these aspects suggest a particular advantage to the viruses of the presence of these sequences.

Animals↗

PubMed1996

Fetal pathology in intrauterine death due to parvovirus B19 infection.

OBJECTIVES: To study the pathological features of fetuses dying because of parvovirus B19 infection, with particular reference to the presence of hydrops; to assess the usefulness of immunochemistry as a screening method for the detection of parvovirus infection at post-mortem examination. DESIGN: Review of clinical, sonographic, serological and pathological data; immunohistochemical staining of post-mortem tissue. SAMPLE: Cases of intrauterine fetal death occurring during the 18-month period January 1993 to June 1994 inclusive, referred for post-mortem examination to the Pathology Department, Royal Victoria Infirmary, Newcastle upon Tyne. RESULTS: Eleven cases of fetal death due to parvovirus infection were identified. Seven fetuses were less than 18-week size. Three fetuses showed conspicuous hydropic change. One of the 11 cases was detected for the first time by retrospective immunochemical screening. Of cases originating from the Newcastle district, parvovirus infection was responsible for about 10% of all non-malformed fetal deaths occurring between 10 and 24 weeks of gestation referred for pathological examination. CONCLUSIONS: During the period of study, parvovirus infection was a relatively common cause of mid-trimester fetal death. Many fetuses dying because of this infection are not noticeably hydropic, and the possibility of parvovirus infection should be considered in any case of intrauterine fetal death. Immunochemistry can be used to confirm the histopathological diagnosis, and may be of particular help where there is advanced autolysis; immunohistochemical screening may detect occasional cases not initially identified by examination of routinely stained tissue sections.

Autopsy↗

PubMed1996

Parvovirus B19 infection and Diamond-Blackfan anaemia.

It is the purpose of the study to report the frequency of parvovirus in children with a diagnosis of Diamond-Blackfan anaemia and to discuss the possible aetiological role of parvovirus in Diamond-Blackfan anaemia. We found parvovirus DNA in 3 of 11 bone marrow smears. Giant pronormoblasts showed low sensitivity (33%) and poor specificity (75%). The presence of giant pronormoblasts was associated with a very high myeloid:erythroid ratio, and may not be specific for parvovirus infection, but a feature of severely suppressed erythropoiesis. The three parvovirus-positive patients were the only children who experienced a remission, and who are free of medication. The seven surviving parvovirus-negative patients are all currently on steroid treatment.

Bone Marrow↗

PubMed1996

Expression of recombinant parvovirus NS1 protein by a baculovirus and application to serologic testing of rodents.

A recombinant baculovirus containing the NS1 gene of minute virus of mice was constructed. Optimal expression of the recombinant NS1 protein (rNS1) was achieved by infecting Trichoplusa ni High Five cells at a multiplicity of 10 and incubating them for 72 h postinfection. An enzyme-linked immunosorbent assay (ELISA) with rNS1 as the antigen was evaluated for serologic testing of laboratory rodents. The rNS1 ELISA proved to be a more sensitive method for the detection of antibodies to recently recognized rodent parvovirus species (mouse orphan parvovirus and rat orphan parvovirus) and prototypic parvovirus species (minute virus of mice, Kilham's rat virus, and H-1) than were conventional parvovirus ELISAs that use whole parvovirus virions.

Amino Acid Sequence↗

PubMed1994

Molecular characterization of a newly recognized mouse parvovirus.

Mouse parvovirus (MPV), formerly known as orphan parvovirus, is a newly recognized rodent parvovirus distinct from both serotypes of minute virus of mice (MVM). Restriction analysis of the MPV genome indicated that many restriction sites in the capsid region were different from those of MVM, but most sites in the nonstructural (NS) region of the genome were conserved. MPV resembled MVM in genome size, replication intermediates, and NS proteins. Replication intermediates in infected cells were the same for MPV and MVM, including packaging of the 5-kb minus (V) strand. Furthermore, the MPV NS proteins were the same size as and present at the same ratio as the MVM(i) proteins in infected cells. Cloning and sequencing of the MPV genome revealed a genome organization closely resembling that of MVM, with conservation of open reading frames, promoter sequences, and splice sites. The left terminal hairpin was identical to that of MVM(i), but the right terminus was not conserved. Also, the MPV genome was unique in that it contained 1.8 copies of the terminal repeat sequence rather than the 1 or 2 copies found in other parvoviruses. The predicted amino acid sequence of the NS proteins of MPV and MVM(i) were nearly identical. In contrast, the predicted amino acid sequence of the capsid proteins of MPV was different from sequences of other parvoviruses. These results confirm that MPV is a distinct murine parvovirus and account for the antigenic differences between MPV and MVM.

Amino Acid Sequence↗

PubMed2001

Recombinant human parvovirus B19 vectors: erythrocyte P antigen is necessary but not sufficient for successful transduction of human hematopoietic cells.

The blood group P antigen, known to be abundantly expressed on erythroid cells, has been reported to be the cellular receptor for parvovirus B19. We have described the development of recombinant parvovirus B19 vectors with which high-efficiency, erythroid lineage-restricted transduction can be achieved (S. Ponnazhagan, K. A. Weigel, S. P. Raikwar, P. Mukherjee, M. C. Yoder, and A. Srivastava, J. Virol. 72:5224-5230, 1998). However, since a low-level transduction of nonerythroid cells could also be detected and since P antigen is expressed in nonerythroid cells, we reevaluated the role of P antigen in the viral binding and entry into cells. Cell surface expression analyses revealed that approximately 75% of primary human bone marrow mononuclear erythroid cells and approximately 31% of cells in the nonerythroid population were positive for P antigen. Two human erythroleukemia cell lines, HEL and K562, and a human promyelocytic leukemia cell line, HL-60, were also examined for P antigen expression and binding and entry of the vector. HEL and K562 cells showed intermediate levels, whereas HL-60 cells demonstrated high levels of expression of P antigen. However, the efficiency of vector binding to these cells did not correlate with P antigen expression. Moreover, despite P antigen positivity and efficient viral binding, HEL, K562, and HL-60 cells could not be transduced with the vector. Low levels of P antigen expression could also be detected in two primary cell types, human umbilical vein endothelial cells (HUVEC) and normal human lung fibroblasts (NHLF). In addition, vector binding occurred in both cell types and was inhibited by globoside, indicating the involvement of P antigen in virus binding to these cells. These primary cells could be efficiently transduced with the recombinant vector. These data suggest that (i) P antigen is expressed on a variety of cell types and is involved in binding of parvovirus B19 to human cells, (ii) the level of P antigen expression does not correlate with the efficiency of viral binding, (iii) P antigen is necessary but not sufficient for parvovirus B19 entry into cells, and (iv) parvovirus B19 vectors can be used to transduce HUVEC and NHLF. These studies further suggest the existence of a putative cellular coreceptor for efficient entry of parvovirus B19 into human cells.

Antigens, Surface↗

PubMed1997

Detection of canine parvovirus in naturally infected dogs with enteritis and myocarditis by in situ hybridization.

An improved method for the diagnosis of canine parvovirus using in situ hybridization in standard formalin-fixed, paraffin-embedded tissue sections was developed. A digoxigenin-labeled probe complementary to DNA sequences that code for the entire sequence of the capsid protein VP-1 and the middle part of the sequence of the capsid protein VP-2 was designed. Specific histologic localization of canine parvovirus-infected cells was demonstrated in small intestine, tonsil, lymph node, thymus, spleen, heart, liver, and kidney from dogs diagnosed at necropsy with canine parvovirus infection. The in situ hybridization accurately pinpointed the specific sites of viral infection. The detection of canine parvovirus in liver, kidney, and heart tissues together in the same pups could represent an enhanced virulence of this strain of canine parvovirus and suggests a broadened tissue tropism not seen before in Korean strains of canine parvovirus.

Animals↗

PubMed2006

Discordant fetal infection for parvovirus B19 in a dichorionic twin pregnancy.

There are only a few reports of fetal parvovirus infection in the circumstance of a twin pregnancy. We report a case of differential fetal infection following maternal parvovirus B19 infection in a dichorionic twin pregnancy culminating in a dual live-birth. A 32-year-old woman was diagnosed with parvovirus infection following exposure to her infected child at 19 weeks' gestation in a dichorionic twin gestation. Maternal IgG seroconversion was documented and maternal blood parvovirus B19 DNA was detected by PCR testing. Fetal monitoring with serial ultrasound assessment was instituted. At 25 weeks' gestation Twin I (male) displayed minor ascites, small pericardial effusion, placentomegaly and oligohydramnios. Middle cerebral artery peak systolic velocity (MCA PSV) studies were abnormal. Twin II (female) was sonographically unremarkable. A single fetal intravascular transfusion for Twin I was performed (pretransfusion Hb 80 g/L). Twin I fetal blood parvovirus B19 DNA was detected by polymerase chain reaction (PCR) and serum demonstrated both B19 IgG and IgM. Following fetal transfusion the hydrops resolved, although the placentomegaly persisted. Two live infants were delivered at 37 weeks' gestation. Individual neonatal venepuncture detected B19 DNA by PCR in Twin I but not in Twin II. B19 IgM was present in Twin I but not Twin II. Both neonates had B19 IgG antibodies. Placental histopathology demonstrated mild edema for Twin I but no inclusion cells. The placenta of Twin II was unremarkable. This case demonstrates the ability for differential transplacental infection of this virus and the assistance of fetal MCA flow studies to guide therapy in a multiple pregnancy at risk of parvovirus infection.

Blood Flow Velocity↗

PubMed1996

A prospective study of antibodies against parvovirus B19 in pregnancy.

BACKGROUND: Parvovirus infection during pregnancy has been reported to be associated with spontaneous abortion and fetal loss. OBJECTIVE: To show the incidence of antibodies against Parvovirus B19 early in pregnancy and sero-reactivities during and after pregnancy. STUDY DESIGN: In a prospective study during a non-epidemic period, 457 women admitted to an antenatal care center were included. Serum samples were collected at 7-13, 21, and 33 weeks of gestation, and 7-9 weeks after delivery. METHOD: Samples were evaluated for parvovirus specific IgG and IgM against two different parvovirus antigens. RESULTS AND CONCLUSION: Parvovirus specific antibodies were present in 81% of the women in the first sample. Six women (six of 88 susceptible, 6.8%) seroconverted and 28 women (28/369, 7.6%) boosted their antibody response during or after pregnancy. All gave birth to healthy infants. One woman free of symptoms experienced an intrauterine fetal death at 37 weeks of gestation. She had no rise in B19 antibodies during pregnancy, but parvovirus DNA was found in maternal serum samples and in the placenta.

Abortion, Spontaneous↗

PubMed2000

Removal of parvovirus B19 from hemoglobin solution by nanofiltration.

Parvovirus B19 (B19), which may contaminate red cell components for blood transfusion, is known to be resistant to several viral inactivation methods. To increase the safety of hemoglobin solutions as a source of red cell substitutes, we investigated the removal of parvovirus B19 from hemoglobin solution using nanofiltration. The hemoglobin solution spiked with parvovirus B19 was tangentially filtered using the BMM-35 filter (mean pore size of 35 nm) followed by BMM-15. The parvovirus B19 of 10(8.5) PT50 (median PCR titer)/10 microL was not changed after the BMM-35 filtration. However, the BMM-15 filtration decreased the parvovirus B19 from 10(8.3-8.7) PT50 to 10(1.3-2.2) PT50, indicating more than 6 log10 reduction. When the initial parvovirus B19 of 10(6.0) PT50 was subjected to the BMM-15 filtration, the residual virus was 10(-0.3-0.5) PT50 orundetected in some fraction of the filtrate. Hemoglobin recovery was 70.4 +/- 3.4%. The ratio of methemoglobin was not changed during the filtration. These findings indicate that the BMM-15 filtration is a promising approach to prepare a safer hemoglobin solution for red cell substitutes.

Blood Substitutes↗

PubMedDate not supplied

Diagnosis and incidence of fetal parvovirus infection in an autopsy series: I. Histology.

This study was undertaken to obtain additional information about the incidence and characteristics of fetal parvovirus B19 infection in an unselected autopsy series, and to assess the distribution and quantitation of inclusions in various organs. Autopsy records from 673 fetal and neonatal autopsies performed at Women and Infants' Hospital during 1985 through 1990 were reviewed. Thirty-two cases of hydrops fetalis were identified, and, of these, 5 had parvovirus infection. This gives an incidence of fetal parvovirus infection resulting in hydrops fetalis of 0.7% among all autopsies, and a 16% incidence among cases of hydrops. Thirty-five percent of the cases of hydrops had malformations; a muscular ventricular septal defect was noted in one of the 5 cases of parvovirus infection. All 5 parvovirus cases had characteristic erythroid nuclear inclusions, and these inclusions were resistant to tissue degenerative changes. The most reliable tissue for histologic diagnosis was the liver, followed by heart and lung. Only 2 of 5 placentas had diagnostic inclusions, making examination of the placenta alone insufficient for ruling out fetal parvovirus infection.

Autopsy↗

PubMedDate not supplied

Diagnosis and incidence of fetal parvovirus infection in an autopsy series: II. DNA amplification.

This study evaluates the practical utility of the polymerase chain reaction (PCR) as a diagnostic method for intrauterine fetal parvovirus infection in cases of hydrops fetalis. Paraffin-embedded, formalin-fixed fetal tissues from cases of hydrops fetalis were assessed for parvovirus B19 by histology and PCR in conjunction with 32P hybridization. Of 673 fetal and neonatal autopsies performed at Women and Infants' Hospital for the years 1985 through 1990, 32 cases were determined to have hydrops fetalis, of which five were positive for parvovirus infection by both histology and the PCR. PCR was not used in seven (22%) of the 32 hydrops cases because 1 microgram of DNA was not available for study. Histology was as sensitive as PCR in detecting parvovirus B19 in fetal autopsy tissues from cases of hydrops fetalis, and could be used reliably in each case to diagnose parvovirus infection. In our hands, histology is as sensitive as PCR and less labor-intensive. We would reserve PCR for cases without inclusions and with a strong suspicion of parvovirus infection, or for fluids in which histological analysis is not available.

Autopsy↗

PubMedDate not supplied

Hydrops fetalis secondary to parvovirus B19 infections.

BACKGROUND: Fetal infection by human parvovirus B19 is a common cause of fetal anemia, nonimmune hydrops fetalis, and spontaneous abortion and can result in fetal death. Recent improvements in diagnosing parvovirus infections and the availability of intrauterine transfusion have reduced the overall rate of fetal loss after maternal exposure. METHODS: We report two cases of maternal parvovirus infection with classic findings of hydrops fetalis and review various aspects of parvovirus infection with emphasis on the developing management options in pregnancy. RESULTS AND CONCLUSIONS: Different management led to different results. In the first case there was normal neonatal and infantile development, and in the second case, the fetus died. With accurate laboratory testing, obstetric sonography, and fetal transfusion, the fetal mortality from parvovirus infection has been reduced considerably, and most pregnancies complicated by maternal parvovirus infection result in healthy outcomes.

Adult↗

PubMed1991

[Blood picture findings in children with Parvovirus B19 infections (fifth disease/erythema infectiosum)].

The human parvovirus B19 provokes erythema infectiosum ("e.i."); moreover there is a wide range of diseases due to parvovirus B19 without exanthema/rash. The erythropoietic blast cells of the bone marrow seems to be the main target cells for this virus. Therefore in cases of prenatal infection the consequences are extremely similar to fetal erythroblastosis ("non-immunological" fetal hydrops). In postnatal life the parvovirus B19 infection causes hyporegenerative phases of the erythropoiesis with anaemia after 3-4 weeks. We studied the white blood cell count (WBC), erythrocytes and thrombocytes in children suffering from (serologically well documented) parvovirus B19 infection with exanthem/"e.i." (group 1; n = 23), without exanthem (group 2; n = 46) and with unknown febrile exanthematous rashes (group 3; n = 76). We did not find any characteristic data in the WBC for a diagnosis of parvovirus B19 infection. However we have for the first time documented a significant thrombocytopenia in "e.i." (group 1) not found in group 2. The thrombocytopenia appears earlier than the anaemia, because the lifespan of thrombocytes is considerably shorter than that of erythrocytes. These data suggest that parvovirus B19 attacks not only "erythropoietic" blast cells but also immature bone marrow cells, which are later responsible for the thrombocytopoiesis.

Adolescent↗

PubMed1994

Chronic parvovirus infection mimicking myelodysplastic syndrome in a child with subclinical immunodeficiency.

PURPOSE: We present a report of a child with subclinical immunodeficiency who became chronically infected with parvovirus resulting in pancytopenia and morphologic abnormalities in the bone marrow mimicking myelodysplastic syndrome (MDS). PATIENTS: An 8-year-old boy presented with severe anemia, moderate thrombocytopenia and granulocytopenia. The patient showed hyper-immunoglobulin M (IgM) immunodeficiency but no increased susceptibility to infections. The bone marrow was hypercellular with dysplastic granulocytopoiesis and erythroblastopenia. RESULTS: Treatment with cyclosporine and i.v. Ig resulted in temporary normalization of the hemoglobin level. For several years it was assumed that the patient had MDS. A diagnosis of parvovirus infection was initially rejected due to the lack of specific antibodies and the absence of giant pronormoblasts in the bone marrow. When the polymerase chain reaction technique became available, parvovirus DNA was detected from the entire disease course. CONCLUSIONS: This case report expands our conception of the clinical spectrum of parvovirus infection and emphasizes that parvovirus must be considered as a differential diagnosis in MDS. We recommend performing a parvovirus DNA test despite negative serologic findings in patients with MDS, especially when associated with immunologic abnormalities.

Child↗

PubMed1997

Evidence for persistence of human parvovirus B19 DNA in bone marrow.

A nested polymerase chain reaction assay (nPCR) was used to investigate the potential of human parvovirus B19 DNA to persist in blood or bone marrow samples obtained either from blood donors or cadaveric bone donors or from patients presenting with clinical signs of parvovirus B19 infection. The presence of parvovirus B19 specific antibody in blood was tested by enzyme immunoassay (EIA). B19 virus genome was not detected in any blood sample of 115 blood donors, of whom 92 (80%) had anti-B19 IgG antibody only as an indication of past infection. In contrast; B19 virus DNA was detected in the bone marrow of 4 out of 45 bone donors. Each one of the serum samples available for 3 of these 4 individuals contained anti-B19 IgG antibody. Among 84 patients with clinical manifestations of parvovirus B19 infection, 17 (20%) had B19 virus DNA in bone marrow. Eight of the latter patients had anti-B19 IgG antibody in their blood but neither anti-B19 IgG nor B19 virus DNA. These data document the ability of parvovirus B19 DNA to persist in the bone marrow of asymptomatic individuals and patients with parvovirus B19 infection suspected on clinical grounds.

Adult↗

PubMed1997

[Human parvovirus B19 infection in perinatal transmission and abnormal fetuses].

OBJECTIVE: To investigate the perinatal infection of human parvovirus B19 in pregnant women and their fetuses. METHODS: 350 sera of normal pregnant women and 9 sera of abnormal fetuses were investigated by polymerase chain reaction (PCR) to check the parvovirus B19 DNA. RESULTS: The prevalence of parvovirus B19 DNA was 1.14% and 0.28% in the sera of normal pregnant women and umbilical cords, respectively. Parvovirus B19 DNA was detected in 6 sera of pregnant women and umbilical cords from 9 abnormal fetuses parvovirus DNA could be detected by in situ hybridization in the nuclei of immature hematopoetic cells within fetal brain or spleen whose PCR tests were positive. CONCLUSION: The results suggest that parvovirus B19 infection do exist in pregnant women and their neonates as well as fetuses in our country. PCR and in situ hybridization could provide more sensitive, more accurate and specific methods for clinical pathologic diagnosis and epidemic investigation.

Adult↗