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PubMed2003

Simultaneous detection and differentiation between porcine circovirus and porcine parvovirus in boar semen by multiplex seminested polymerase chain reaction.

A multiplex seminested polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among porcine circovirus 1 (PCV1), PCV2, and porcine parvovirus (PPV) from boar semen. Primers of PCV1, PCV2 and PPV were specific and did not react with other viruses respectively. Twenty (20.4%) and 42 (42.9%) out of 98 whole semen samples were found to be positive for PCV and PPV using multiplex conventional and seminested PCR, respectively. When the separated fractions of PCV or PPV-contaminated semen were analyzed using multiplex seminested PCR, PCV and PPV DNA were found to be present mainly in the seminal fluid and nonsperm cell fractions. This multiplex seminested PCR assay was sensitive, rapid and a good alternative method for the detection and differentiation of these viruses in boar semen.

Animals↗
PubMed2005

VP2 gene of a canine parvovirus isolate from stool of a puppy.

VP2 gene of a canine parvovirus (CPV) isolate from the feces of a puppy which was diagnosed to be CPV infection was analysed. The result indicated that this clinical isolate was phylogenetically close to the isolate of wild-type CPV (strain CPV-T37) prevailing in Taiwan rather than isolates from Japan.

Amino Acid Sequence↗
PubMed2000

Human parvovirus B19 in pregnancy.

Parvovirus B19 infection can result in an adverse outcome when acquired during pregnancy. However, in the majority of cases a successful outcome can be anticipated. Public awareness of this condition is essential and obstetricians should be familiar with the options available to them if they are presented with this clinical problem.

Anemia↗
PubMed1999

Subacute massive necrotizing myocarditis by canine parvovirus type 2 infection with diffuse leukoencephalomalacia in a puppy.

Severe necrotizing myocarditis associated with canine parvovirus type 2 (CPV-2) infection and diffuse leukoencephalomalacia was recognized in a 4-week-old mongrel puppy. The cardiac lesions were characterized by severe diffuse myocardial degeneration and necrosis with occasional massive mineralization and distinct intranuclear inclusion bodies. Immunohistochemical examination revealed the presence of CPV-2 antigens in both the cytoplasm and nuclei of the myocytes. In the white matter of the cerebrum, moderate to severe diffuse necrosis with diffuse infiltration of gitter cells and occasional diffuse mineral deposits were recognized symmetrically and bilaterally. In the meningocortical area of the cerebellum, there was mild focal hemorrhage and accumulation of hemosiderin-laden histiocytes. In addition to the absence of viral antigen (as judged by immunohistochemistry), neither viral inclusions nor other vascular lesions were identified in examined sections of brain. These findings suggest that the brain lesions were not induced by direct CPV-2 infection but were related to severe myocardial lesions producing prolonged hypoxia and/or ischemia.

Animals↗
PubMed2000

Viral wasting syndrome of swine: experimental reproduction of postweaning multisystemic wasting syndrome in gnotobiotic swine by coinfection with porcine circovirus 2 and porcine parvovirus.

One-day-old gnotobiotic piglets were inoculated intranasally with in vitro passaged porcine circovirus 1 (PCV-1), PCV-2, and porcine parvovirus (PPV) alone or in combination (PCV-1/PCV-2, PCV-1/PPV, and PCV-2/PPV). Piglets were evaluated for 1) the development of porcine postweaning multisystemic wasting syndrome (PMWS), 2) distribution of viral antigens by immunochemistry, and 3) viremia and the presence of viral DNA in nasal and ocular secretions and feces. All single agent-infected piglets and piglets infected with PCV-1/PCV-2 or PCV-1/PPV were clinically asymptomatic. They were transiently viremic and seroconverted to homologous virus(es). At termination of the study on postinfection day (PID) 35, microscopic lesions were restricted to focal inflammatory cell infiltrates in livers and myocardia. One piglet given PCV-1/PPV was PPV viremic for 2 weeks after infection and had lymphangiectasia of the spiral and descending colon associated with granulomatous inflammation. All four PCV-2/PPV-inoculated piglets developed PMWS, characterized by sudden onset of depression and anorexia, icterus, and submucosal edema. One piglet became moribund on PID 27, and the remaining three piglets were euthanatized between PID 27 and PID 30 because of severe disease. Lymph nodes were small and the livers were mottled. Disseminated angiocentric granulomatous inflammation was present in all tissues examined except the brain. Multiple lightly basophilic intracytoplasmic inclusion bodies were identified in macrophages and histiocytes. PCV-2 antigen was widely distributed within macrophages; PPV antigen was sparse. Hepatocellular necrosis and bile retention were prominent. PCV-2 DNA was identified in ocular, fecal, and nasal secretions. Terminal sera contained antibodies to PPV (4/4) and PCV-2 (3/ 4). Production of PMWS in gnotobiotic swine appears to require PCV-2 and additional infectious agents such as PPV for full disease expression in gnotobiotic piglets.

Animals↗
PubMed2006

Potentiation of porcine circovirus 2-induced postweaning multisystemic wasting syndrome by porcine parvovirus is associated with excessive production of tumor necrosis factor-alpha.

This study investigated the potentiation of porcine circovirus 2 (PCV2)-induced postweaning multisystemic wasting syndrome by porcine parvovirus (PPV) and found it was associated with excessive production of tumor necrosis factor-alpha (TNF-alpha). Colostrum-deprived conventional pigs were inoculated intranasally with PCV2 or PPV alone or in combination (PCV2 and PPV). In vitro assay of TNF-alpha, obtained from alveolar macrophages coinfected with PCV2 and PPV, showed a significant increase in TNF-alpha compared to single infection of macrophages with either PCV2 or PPV alone (P < 0.05). All pigs inoculated with PCV2 and PPV developed severe postweaning wasting syndrome, whereas clinical signs (e.g., weight loss) were present but perhaps less severe in either PCV2- or PPV-inoculated pigs. Compared to the pigs inoculated with PCV2 or PPV alone, pigs inoculated dually with PCV2 and PPV showed significantly (P < 0.05) increased levels of TNF-alpha. Levels of TNF-alpha in the sera were reversely correlated with the body weight in pigs experimentally infected with dual inoculation of PCV2 and PPV (r(s) = -0.92, P < 0.001). These data suggest that a potentiation of PPV in PCV2-induced PMWS is associated with the excessive production of TNF-alpha.

Animals↗
PubMed2006

Peptide nucleic acid-based in situ hybridization assay for detection of parvovirus B19 nucleic acids.

BACKGROUND: Peptide nucleic acid (PNA) molecules are known to bind complementary nucleic acid sequences with a much stronger affinity and with more stable binding than DNA or RNA molecules. We chose parvovirus B19, which is diagnosed by detection of nucleic acids by in situ hybridization assay (ISH) and/or PCR, as an experimental model to develop an ISH assay that uses biotinylated PNA probes to detect viral genome in clinical specimens. METHODS: We first optimized the PNA-ISH assay on B19-infected and mock-infected UT-7/EpoS1 cells and then tested the assay on archival B19 specimens and on consecutive specimens. All data were compared with data obtained with a standardized DNA-based ISH assay and confirmed by a PCR-ELISA. RESULTS: PNA-ISH detected B19 genome in a higher number of B19-infected UT-7/EpoS1 cells and with a more defined localization of viral nucleic acids than the standardized DNA-ISH assay. Moreover, PNA-ISH was able to detect B19 genome in all positive archival samples, whereas DNA-ISH failed in 5 samples. PNA-ISH detected more positive samples than DNA-ISH when consecutive specimens were analyzed, and a close agreement was found with PCR-ELISA results. CONCLUSIONS: The PNA-ISH assay had sensitivity and specificity comparable to a PCR assay and was more practical and quicker to perform than standard hybridization assays. The assay may be a suitable diagnostic test for the detection of viral nucleic acids in clinical specimens.

Cell Line↗
PubMed2004

Monitoring human parvovirus B19 virus-like particles and antibody complexes in solution by fluorescence correlation spectroscopy.

Fluorescence correlation spectroscopy (FCS) was used in monitoring human parvovirus B19 virus-like particle (VLP) antibody complexes from acute phase and past-immunity serum samples. The Oregon Green 488-labeled VLPs gave an average diffusion coefficient of 1.7 x 10(-7) cm2 s(-1) with an apparent hydrodynamic radius of 14 nm. After incubation of the fluorescent VLPs with an acute phase serum sample, the mobility information obtained from the fluorescence intensity fluctuation by autocorrelation analysis showed an average diffusion coefficient of 1.5 x 10(-8) cm2 s(-1), corresponding to an average radius of 157 nm. In contrast, incubation of the fluorescent VLPs with a past-immunity serum sample gave an average diffusion coefficient of 3.5 x 10(-8) cm2 s(-1) and a radius of 69 nm. A control serum devoid of B19 antibodies caused a change in the diffusion coefficient from 1.7 x 10(-7) to 1.6 x 10(-7) cm2 s(-1), which is much smaller than that observed with acute phase or past-immunity sera. Thus, VLP-antibody complexes with different diffusion coefficients could be identified for the acute phase and past-immunity sera. FCS measurement of VLP-immune complexes could be useful in distinguishing between antibodies present in acute phase or past-immunity sera as well as in titration of the VLPs.

Animals↗
PubMed2005

Intrauterine therapy for parvovirus B19 infected symptomatic fetus using B19 IgG-rich high titer gammaglobulin.

Parvovirus B19-infected hydrops fetalis was treated using gammaglobulin injection into the peritoneal cavity (GIFPeC) with B19-IgG-rich immunoglobulin. Fetal anemia and hydrops resolved, and B19-DNA in fetal ascites decreased despite no change in maternal B19-IgG or B19-DNA. Gammaglobulin injection into the peritoneal cavity is thus useful for treating hydrops fetalis while avoiding intrauterine blood-transfusion risks.

Adult↗
PubMed1994

The frequency of human parvovirus B19 infection in nonimmune hydrops fetalis.

Intrauterine infection of human parvovirus B19 has recently been identified as an etiology for nonimmune hydrops fetalis (NIHF) and fetal death. To examine the frequency of B19 infection in cases of NIHF, forty two cases of NIHF of unknown cause were tested for the presence of B19 antibodies by enzyme-linked immunosorbent assay (ELISA) and B19 DNA by polymerase chain reaction (PCR). Three maternal sera were positive for B19 IgM antibody and one fetal serum was positive for B19 DNA. Overall, four (10%) of the 42 cases were positive for B19 infection. All of the four cases positive for B19 infection were found during or just before an outbreak of erythema infectiosum and the incidence was calculated at 36% if only cases found during the outbreak were collected. In all four cases of B19 infection, NIHF was diagnosed at 20 to 23 weeks of gestation, which suggested that B19 infection might be a particular threat to the fetus during this stage of gestation. In conclusion, B19 infection may contribute to 10% of cause unknown NIHF and the incidence may be higher if cases during outbreaks are exclusively collected.

Antibodies, Viral↗
PubMed1994

Fetoneonatal hydrops from human parvovirus B19. Case report.

Authors report the case of a newborn who died just a few hours after the birth as a result of intrauterine Parvovirus B19 infection. Diagnosis of fetal hydrops was made by ultrasound examination at the 25th week of pregnancy. Etiology was established on the basis of specific antibody findings in the serum of the mother, the fetus (by cordocentesis), and the neonate; B19 virus was then observed in the fetus and the neonate tissues after death using the dot-blot hybridization assay and the polymerase chain reaction technique for viral DNA. The severe fetal anemia was treated with intrauterine transfusions, but achieved poor results. The pathogenesis of fetal hydrops and advisability of intrauterine treatment in such cases are discussed.

Adult↗
PubMed2004

Successful bone marrow transplantation for severe aplastic anemia in a patient with persistent human parvovirus B19 infection.

Persistent infection with human parvovirus B19 (B19) is primarily associated with chronic bone marrow failure in immunocompromised patients, but occasionally this organism may also affect immunocompetent hosts. B19 is also suggested as a causative agent of organ failure during bone marrow transplantation (BMT). We herein report the case of a 9-year-old girl with no previous history of immunodeficiency who developed severe aplastic anemia concurrent with B19 persistent infection. Both immunoglobulin (Ig)M antibody to B19 and B19 DNA identified by real-time polymerase chain reaction were found in the patient's serum at time of diagnosis of aplastic anemia. No giant proerythroblasts were found in her bone marrow at diagnosis. Although intravenous administration of Ig (IVIg) reduced serum B19 DNA, the aplastic status of her bone marrow did not improve. Both aplastic anemia and persistent B19 viremia were successfully treated by BMT from an HLA-identical sibling donor. Serum B19 DNA increased temporarily after BMT; however, neither organ nor marrow failure was observed. B19 DNA disappeared from the serum 2 months after BMT, suggesting that a normal immune response was restored by BMT and terminated the B19 viremia. During BMT, use of high-titer IVIg for B19 might prevent B19-associated organ failure.

Anemia, Aplastic↗
PubMed2004

Successful treatment with cyclosporine and high-dose gamma immunoglobulin for persistent parvovirus B19 infection in a patient with refractory autoimmune hemolytic anemia.

We describe a patient with persistent pure red cell aplasia due to human parvovirus B19 (HPVB19) infection during immunosuppressive therapy for refractory autoimmune hemolytic anemia (AIHA). The patient had been given corticosteroid (CS) and/or azathioprine for AIHA. During the course of treatment, reticulocyte count and hemoglobin levels decreased suddenly. Bone marrow aspirate showed erythroid lineage-specific aplasia with a few giant proerythroblasts, suggesting the presence of HPVB19 infection. The diagnosis of aplastic crisis due to HPVB19 infection was based on positive test results by polymerase chain reaction for HPVB19 immunoglobulin M (IgM) antibody and B19 DNA. Although splenectomy followed by administration of high-dose gamma globulin (HDIG) and plasma exchange were performed, the crisis and hemolysis recurred. Aplastic crises occurred several times when the B19 IgG result became negative and the CD4+ lymphocyte count was less than 300/microL. The patient showed complete recovery from anemia after CS was switched to cyclosporin A (CsA) and intermittent administration of HDIG. The result for B19 IgG antibody was continuously positive, and the DNA result became negative after these treatments. The results in this case indicated that concomitant administration of CsA and intermittent administration of HDIG can lead to cure of chronic anemia due to HPVB19 infection in patients with refractory AIHA.

Adult↗
PubMedDate not supplied

Papular-purpuric "gloves and socks" syndrome due to parvovirus B19: report of a case with unusual features.

We present a case of papular-purpuric "gloves and socks" syndrome (PPGSS) in an adult male with acute parvovirus B19 infection. The patient displayed the classical features of fever, oral lesions, and purpura on hands and feet, but the purpuric lesions on the feet evolved to superficial skin necrosis, a feature not previously described in this syndrome. We believe this is the first reported case of PPGSS occurring in Brazil.

Acrodermatitis↗
PubMedDate not supplied

Human parvovirus B19 infections among exanthematic diseases notified as measles.

A total of 1397 sera collected from 1095 cases of exanthematic disease notified as measles in ES and RJ states during July 1992 to December 1994 were investigated. These sera were first tested for measles and rubella specific IgM. When they proved negative, they were tested for B19 specific IgM by an enzyme immunoassay. B19 infection was confirmed in 27 (2.5%) of these cases. Sera from 194 negative cases for measles and rubella IgM received from other Brazilian states were also investigated and B19 infection was confirmed for 11 of them. Sera from these 38 IgM positive cases for B19, were tested for anti-B19 IgG by an enzyme immunoassay and for B19 DNA by dot blot hybridization. Anti-B19 IgG antibodies were detected in most of the acute sera. B19 DNA was detected in the acute serum of one patient that had been splenectomized before. As the exanthem caused by human parvovirus infection may be clinically diagnosed as rubella, it could be important to diagnose B19 infection in Brazil since it is becoming prevalent as the cause of rash in countries where rubella is controlled by vaccination.

Antibodies, Viral↗
PubMedDate not supplied

Human parvovirus B19 infection in HIV-positive patients.

Parvovirus B19 infects predominantly erythroid cells, leading to transient inhibition of erythropoiesis. Immunocompromised patients may be unable to produce neutralizing antibodies and may develop severe chronic anemia. Epidemiological studies done on Niterói population showed that B19 infection occurs periodically in late spring and summer. We report a study from 55 HIV infected patients attending an infectious diseases outpatient clinic in this city during a 5-month period in which B19 circulation was well documented. All patients were under anti-retroviral therapy. No anti-B19 IgM was found, but a high prevalence of IgG anti-B19 (91%) was observed. In six patients, B19 DNA was found by dot-blot hybridization techniques, but this was not confirmed by PCR. None of these 6 patients manifested anemia and only one had CD4 cell count below 200 x 10(7)/L. We conclude that persistent infection causing anemia is an infrequent finding in our HIV positive patients under drug therapy.

Adult↗
PubMedDate not supplied

Human parvovirus B19 infection and hydrops fetalis in Rio de Janeiro, Brazil.

Formalin-fixed paraffin embedded lung and liver tissue from 23 cases of non immune hydrops fetalis and five control cases, in which hydrops were due to syphilis (3) and genetic causes (2), were examined for the presence of human parvovirus B19 by DNA hybridisation. Using in situ hybridisation with a biotynilated probe one positive case was detected. Using 32P-labelled probes in a dot blot assay format, five further positives were obtained. These were all confirmed as positive by a nested polymerase chain reaction assay. Electron microscopy revealed virus in all these five positive cases. The six B19 DNA positive cases of hydrops fetalis were from 1974, 1980, 1982, 1987 and 1988, four of which occurred during the second half of the year, confirming the seasonality of the disease.

Brazil↗
PubMed2001

Anti-VP1 and anti-VP2 antibodies detected by immunofluorescence assays in patients with acute human parvovirus B19 infection.

Acute human parvovirus B19 infection is followed by an antibody response to the structural proteins of the viral capsid (VP1 and VP2). We used 80 sera collected from 58 erythema infectiosum and 6 transient aplastic crisis patients to test IgM and IgG antibodies against these two proteins in an immunofluorescence assay (IFA) using Sf9 cells infected with recombinant baculovirus expressing either VP1 or VP2 antigen. Although less sensitive than IgM capture enzyme immunoassay using native antigen (MACEIA), we could detect anti-VP1 or anti-VP2 IgM antibodies by IFA in 49 patients with acute infection (76.6%). Detection of IgG anti-VP1 and anti-VP2 by IFA, however, was as sensitive as IgG detection by indirect enzyme immunoassay. By applying IgG avidity IFA to sera of the 15 IgM IFA negative patients we were able to confirm acute infection in further 12 cases by IFA. Overall, acute infection was confirmed by IFA in 61 (95.3%) of the 64 patients.

Adolescent↗

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