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PubMed1995

Genetic diversity in the non-structural gene of parvovirus B19 detected by single-stranded conformational polymorphism assay (SSCP) and partial nucleotide sequencing.

A homologous region in the parvovirus B19 non-structural gene (B19 nt 1399-1682) was examined in 50 samples from patients with a wide variety of B19-related disease from various countries by PCR amplification, single-stranded conformational polymorphism (SSCP) assay and nucleotide sequencing. Five SSCP types were confirmed by nucleotide sequence analysis. Of a total of 6 mutations, all were silent. Types 3 and 4 accounted for 92% of strains. There was no correlation between genome type and either clinical illness or patient age. However, there was a correlation between SSCP type and country of origin. Type 3 strains predominated in Japan (18/26) and the UK (6/8), whereas type 4 predominated in the USA (9/12). Notably, type 3 strains also predominated among females (14/18), whereas there were approximately equal numbers of strain types 3 (7/17) and 4 (8/17) among males; an observation which remains unexplained. Within the Japanese group, although type 3 strains predominated overall, strains isolated from 1981 to 1987 consisted of types 1 (2/15), 2 (1/15), 3 (8/15), and 4 (4/15), whereas strains isolated from 1990 to 1994 consisted almost entirely of type 3 (10/11).

Adolescent↗
PubMed1995

Evaluation of 4 commercial test kits for parvovirus B19-specific IgM.

Four commercial test kits for parvovirus B19 IgM were evaluated by testing 491 sera assembled into 7 panels. The serum panels were designed to assess sensitivity and specificity of the commercial assays and to reflect the various clinical settings in which acute B19 infection forms part of the differential diagnosis. A mu-capture radioimmunoassay (MACRIA) was used as the reference test. With respect to MACRIA, the commercial B19 IgM assays showed an overall sensitivity of 70.1-84.1% and specificity of 92.2 to 97.4%. Assay performance varied in different clinical situations. In sera from adults with acute B19 arthropathy, all 4 assays were 100% sensitive, but in children with fifth disease, the sensitivity ranged from 44.1 to 88.6%. The sensitivity of all 4 assays was also low when testing samples collected more than 6 weeks after onset of symptoms and in women with B19-associated embryopathy. Specificity was greater than 97% in healthy blood donors, but varied from 70.9 to 83.3% in patients acutely infected with other viruses, including rubella. Although the IgM test kits here evaluated may be usefully introduced for B19 diagnosis in certain settings, knowledge of their limitations will be important when results have been interpreted.

Acute Disease↗
PubMed1995

A synthetic parvovirus B19 capsid protein can replace viral antigen in antibody-capture enzyme immunoassays.

To establish a renewable source of parvovirus B19 antigens for diagnostic tests, gene sequences for the viral capsid proteins, VP1 and VP2, were cloned into baculovirus expression vectors and the recombinant viruses used to infect Sf9 insect cells. Cell lysates examined by immunoblotting demonstrated reactive proteins corresponding to the expected sizes of native VP1 (83 kDa) and VP2 (58 kDa). The VP2 protein was produced efficiently in quantity and self-assembled into empty capsids as shown by density equilibration in a CsCl step gradient. The VP2 protein was purified and used as an antigen in antibody-capture enzyme immunoassays for the detection of B19 IgG and IgM antibodies. Compared to a standard antibody-capture EIA based on whole viral antigen, the VP2-EIA gave a sensitivity of 100% and specificity of 97% in detection of B19 IgM in 138 patients suspected of B19 infection. No IgM-positive specimens were missed. IgG detection yielded a sensitivity of 100% and specificity of 96% in the same population. Recombinant VP2 capsid proteins expressed in baculovirus-infected insect cells can substitute for serum-derived B19 virus in standard antibody-capture EIA for the detection of B19 IgG and IgM with comparable results.

Adult↗
PubMed1992

Petechial glove and sock syndrome caused by parvovirus B19.

The petechial glove and sock syndrome is a recently described febrile dermatosis characterized by acral pruritus, edema, pain, petechiae, and an enanthem of petechiae and erosions; these features suggest a viral origin. We report a typical case in a 36-year-old woman. IgM antibodies to human parvovirus B19 (PVB19) were present, and acute and convalescent IgG antibodies demonstrated seroconversion, which suggested recent infection with PVB19. Results of tests for other viral and bacterial agents were negative. These results strongly implicate PVB19 as an etiologic agent in the petechial glove and sock syndrome.

Acute Disease↗
PubMed1994

Evaluation of a novel diagnostic test for canine parvovirus.

The CPV ELISA detection kit (CSL Ltd., Melbourne Australia) was developed as a rapid field test for the detection of canine parvovirus (CPV) antigen in canine faecal samples. The kit utilises a novel concept in enzyme immunoassay (ELISA) where the conjugate is lyophilised along with the solid-phase capture antibody in the test well. The CPV ELISA was compared to the haemagglutination assay (HA) test using electron microscopy (EM) and/or virus isolation (VI) to confirm infection. The CPV ELISA had a sensitivity of 87% and a specificity of 100% compared to 87% and 63% respectively for the HA. The poor specificity of the HA results in a low positive predictive value of 51% compared to 100% for the CPV ELISA. The CPV ELISA requires no specialised equipment, is simple to perform and provides a visible result in less than 15 minutes.

Animals↗
PubMed1994

Sensitive detection of canine parvovirus DNA by the nested polymerase chain reaction.

A polymerase chain reaction (PCR) for the detection of canine parvovirus (CPV) was developed. To increase the sensitivity and specificity of the reaction, the nested PCR with a double-nested primer pair (inner primer pair) was designed. The sequences of the PCR primer pairs were selected from the conserved region in the CPV VP1/VP2 gene. The PCR with the outer or inner primer pair alone (single PCR) could detect 10 fg of viral replicative form (RF) DNA on agarose gel electrophoresis; whereas as little as 100 ag of the RF DNA was detected by the nested PCR, which was shown to be 100 times more sensitive than the single PCR. Samples prepared from feline panleukopenia virus and mink enteritis virus, both having a very close antigenic relationship to CPV, were also amplified by the nested PCR. The specificity of the reaction was confirmed by restriction enzyme analysis and Southern hybridization. Next, fecal samples were examined by the nested PCR. All 10 samples suspected of CPV infection were positive, and two restriction sites (HaeIII and HindIII sites) in the PCR product were conserved among them. On the other hand, specific amplification was not observed in the samples derived from normal dogs. The number of the genome copy in positive samples was estimated about 10(9)-10(11)/g by the single PCR and 10(11)-10(13)/g by the nested PCR. The assay can be completed in 1-1.5 days, and does not need radioisotopes. Thus, the nested PCR seems to be a sensitive, specific and practical method for the detection of CPV in fecal samples.

Animals↗
PubMed1995

Diagnosis of fetal infection with porcine parvovirus by in situ hybridization.

In situ hybridization (ISH) for the diagnosis of fetal infection with porcine parvovirus (PPV) was compared with immune electron microscopy (IEM) and serology by immunofluorescence (IF) for its sensitivity and its applicability in a routine diagnostic laboratory. The technique was applied to the examination of sections of formalin-fixed paraffin-embedded tissues from 68 fetuses. Fifty-three of these fetuses were diagnosed serologically since they had a crown rump length of more than 17 cm, i.e. they were mature enough to mount a humoral immune response; 38 were positive and 15 negative. Eleven out of 15 smaller fetuses examined for the presence of viral antigen by immune electron microscopy (IEM) were positive and 4 were negative. Heart and lung were found to be the most suitable organs for in situ hybridization. In situ hybridization yielded a positive result in 8 of the 11 IEM positive fetuses and in 33 of the 38 serologically positive fetuses. No signal was detected in any of the 4 IEM or the 13 serologically negative fetuses. Expenses for IEM were estimated to be 179% of the expenses for ISH. Expenses for serology by IF on the other hand were 67% of the expenses for ISH. From this it was concluded that the most efficient way to diagnose a fetal infection with PPV was serology by IF, if possible with samples from several fetuses and that the other techniques, IEM or ISH, ought to be reserved for those cases where no immunocompetent fetuses were available for diagnosis.

Animals↗
PubMed1996

Canine parvovirus binds to multiple cellular membrane proteins from both permissive and nonpermissive cell lines.

For identification of canine parvovirus (CPV) binding protein, the SDS-solubilized cell membrane fraction from a permissive cell line. CRPK, was subjected to the virus overlay protein blot assay (VOPBA). Competitive inhibition experiments showed the presence of multiple CPV-binding proteins with molecular masses of 36, 35, 33, 31, 29, 27, 25, and 23 kDa. CPV-binding proteins of same molecular masses were also detected in membrane fractions from nonpermissive, as well as other permissive, cell lines. We confirm that the mechanism of nonpermissiveness to CPV is not operative at the cellular attachment level.

Animals↗
PubMed2004

Intravenous immunoglobulins in autoimmune- or parvovirus B19-mediated pure red-cell aplasia.

Pure red-cell aplasia (PRCA) is defined as the absence of mature erythroid precursors in a bone marrow that otherwise exhibit normal cellularity. Acquired PRCA may occur in association with neoplasms (such as lymphoproliferative disorders), thymoma, autoimmune disorders, pregnancy, or as a consequence of chronic human parvovirus B19 (B19) infection in an immunologically incompetent host. PRCA may also develop after exposure to drugs (erythropoietin or tacrolimus). PRCA of autoimmune origin was first treated successfully with intravenous immunoglobulins (IVIg) more than 20 years ago. Since then, B19-associated PRCA in solid-organ transplant recipients and in human immunodeficiency virus (HIV)-infected patients has also been successfully treated with IVIg. Routine maintenance therapy is probably not indicated in HIV-infected patients with CD4+ counts above 300/microL, whereas repeated infusions might be necessary if CD4+ count is below 80.

Autoimmune Diseases↗
PubMed2006

Unscrambling the role of human parvovirus B19 signaling in systemic autoimmunity.

Despite enormous progress in understanding how the immune system works, the pathogenesis of autoimmune diseases still remains unclear. Growing evidence indicates that infectious agents can be potent initial triggers, subverting and exploiting host cell signaling pathways. This role is exemplified by the association of parvovirus B19 (B19) with human autoimmune disease. Infection with this common virus exhibits striking similarities with systemic autoimmune diseases, and can be associated with elevated serum autoantibody titers. The B19 virus produces proline-rich, 11-kDa proteins that have been implicated in modulation of host signaling cascades involved in virulence and pathogenesis. Additionally, B19 produces a non-structural protein (NS1) that functions as a transcription regulator by directly binding the p6 promoter and the Sp1/Sp3 transcription factors. The protein is also involved in DNA replication, cell cycle arrest and initiation of apoptotic damage, particularly in erythroid cells. When transfected to non-permissive cells, NS1 recruits the mitochondria cell death pathway. It is even more remarkable that NS1 functions as a trans-acting transcription activator for the IL6 promoter, up-regulating IL6 expression in host cells. Hence, B19 infection may play a pivotal role in triggering inflammatory disorders. By promoting apoptotic damage and trans-activating pro-inflammatory cytokine promoters, B19 may break the delicate balance between cell survival and apoptosis, and may contribute to immune deregulation. Understanding the mechanisms used by B19 to alter the cell signaling machinery may provide further insight into the mechanism by which autoimmune diseases develop.

Apoptosis↗
PubMed2005

Maternally-derived antibodies in pups and protection from canine parvovirus infection.

The interaction between maternally-derived antibodies (MDA) and canine parvovirus (CPV) infection was evaluated in five groups of pups with a wide range of haemagglutination inhibiting (HI) titres of MDA (from < 10 to 320). The pups were inoculated with a field CPV strain and monitored daily to evaluate their clinical condition and viral shedding in the faeces. Serum samples were collected weekly to evaluate antibody response. Clinical signs were observed in dogs with HI titres up to 80. Active CPV replication was demonstrated in dogs with HI titres up to 160, although slightly delayed, at lower titres and for a shorter period compared to seronegative dogs. The successful infection of dogs with HI titres of 80 and 160 was also confirmed by seroconversion, evaluated at day 14 postinfection. These findings are in contrast with the MDA titre (HI > or = 80) usually considered fully protective for CPV infection, and suggest the need for revision of current vaccination programmes for pups.

Animals↗
PubMed2006

The association of VP1 unique region protein in acute parvovirus B19 infection and anti-phospholipid antibody production.

BACKGROUND: Previous studies have postulated a connection between human parvovirus B19 (B19) infection and anti-phospholipid antibodies (aPL). Recently, the phospholipase domain of B19 has been linked to B19-VP1 unique region (VP1u). To elucidate the roles of VP1u in B19 infection and aPL production, the major reactivity of anti-B19-VP1u, anti-cardiolipin antibody (aCL), and anti-beta2-glycoprotein I (beta2GPI) antibody was evaluated. METHODS: Sera from 102 clinically suspected cases of B19 infection were analyzed by nested PCR and ELISA. Humoral responses of anti-B19-VP1u and anti-B19-VP1uD175A IgM/IgG antibodies, aCL and the anti-beta2GPI antibody were assessed by Western blot and ELISA. Absorption experiments were also performed to determine the binding specificity of immunoglobulins to B19-VP1u, CL and beta2GPI. RESULTS: Sera from patients with the diagnostic pattern DNA+/IgM+/IgG+ had a high frequency (57%) for recognition of CL and beta2GPI. Furthermore, adsorption experiments were performed by adding purified B19-VP1u, which partially suppressed the reactivity of anti-B19VP1u to CL and beta2GPI. CONCLUSIONS: Serum from patients with acute B19 infection has a high frequency in recognition of CL and beta2GPI, and the phospholipase domain observed in the B19-VP1u may have contributed to the production of aPL. These findings may provide a clue for understanding the roles of B19-VP1u in B19 infection and aPL production.

Adolescent↗
PubMed2006

Sequence analysis of VP2 gene of canine parvovirus isolated from domestic dogs in Japan in 1999 and 2000.

Seven strains of canine parvovirus (CPV) were isolated from affected dogs in Japan between 1999 and 2000, and their VP2 genes were genetically analyzed. Comparison of the predicted amino acid sequences of VP2 suggested that three field isolates corresponded to CPV type 2a, while the other four to CPV type 2b. The phylogenetic tree constructed from the VP2 genes showed that the newly isolated strains are classified into the cluster consisting of recent Japanese and Taiwanese field isolates, which are distinct from Vietnamese isolates, United States Isolates, or classical CPV type 2. These results suggest that the CPV transmission occurred between Japan and Taiwan in 1990s, and the offspring are still circulating in both countries.

Amino Acid Sequence↗
PubMed2005

Diagnosis and management of human parvovirus B19 infection.

Parvovirus B19 is a common infection in children occurring in winter and spring months that can affect pregnant women and their unborn fetuses. Adverse fetal sequelae may include red blood cell aplasia due to bone marrow suppression, anemia, myocarditis, nonimmune hydrops, and fetal death. Fortunately, several noninvasive testing schemes are available to assess the fetus, and invasive testing is reserved for fetuses with marked anemia.

Antibodies, Viral↗
PubMed2005

Expression and subcellular targeting of canine parvovirus capsid proteins in baculovirus-transduced NLFK cells.

A mammalian baculovirus delivery system was developed to study targeting in Norden Laboratories feline kidney (NLFK) cells of the capsid proteins of canine parvovirus (CPV), VP1 and VP2, or corresponding counterparts fused to EGFP. VP1 and VP2, when expressed alone, both had equal nuclear and cytoplasmic distribution. However, assembled form of VP2 had a predominantly cytoplasmic localization. When VP1 and VP2 were simultaneously present in cells, their nuclear localization increased. Thus, confocal immunofluorescence analysis of cells transduced with the different baculovirus constructs or combinations thereof in the absence or presence of infecting CPV revealed that the VP1 protein is a prerequisite for efficient targeting of VP2 to the nucleus. The baculovirus vectors were functional and the genes of interest efficiently introduced to this CPV susceptible mammalian cell line. Thus, we show evidence that the system could be utilized to study targeting of the CPV capsid proteins.

Animals↗
PubMed2004

Sudden cardiac death in a 5-year-old girl associated with parvovirus B19 infection.

We report on a 5-year-old girl who suddenly collapsed and died while dancing at a family party. Histological examination of the heart including the cardiac conduction system revealed lymphocytic infiltrations of the sinu-atrial node and perivascular infiltration in the atrio-ventricular region. Additionally, foci of mononuclear infiltrates were observed in the myocardium. Consequently, myocarditis was diagnosed as cause of death. The child also had lymphocytic conjunctivis, parotitis and tracheitis. Evaluation of infections by means of nested polymerase chain reaction revealed parvovirus B19 DNA (PVB19) in tissue samples of the trachea.

Child, Preschool↗
PubMed2006

Tako-Tsubo cardiomyopathy (apical ballooning) with parvovirus B19 genome in endomyocardial biopsy.

The phenomenon of transient apical ballooning is a rare underlying cause of severe left ventricular dysfunction and has been described as Tako-Tsubo-like cardiomyopathy. Acute myocarditis has been reported to masquerade as acute myocardial infarction or vice versa and is considered as differential diagnosis in this phenomenon. We present here a case of an adult female who was admitted to our cardiology department with chest pain, electrocardiographic and echocardiographic features, suggestive of an acute anterior myocardial infarction, preceded by physical and emotional stress. Coronary angiography demonstrated coronary arteries without relevant atherosclerotic lesions; left ventriculography showed a severe anteroapical dysfunction. Right ventricular endomyocardial biopsy showed normal cardiac tissue, but was positive for Parvovirus B19. Follow-up (2 months later) showed complete regression of regional wall motion abnormalities in transthoracic echocardiography.

Aged↗
PubMed2006

Presence of perineuritis in a case of papular purpuric gloves and socks syndrome associated with mononeuritis multiplex attributable to B19 parvovirus.

A previously healthy 16-year-old boy developed papular purpuric gloves and socks syndrome concomitant with mononeuritis multiplex attributable to acute infection with parvovirus B19. A cutaneous biopsy showed lymphocytic lichenoid vasculitis and perineuritis. We discuss the relevance of these pathologic findings, and we suggest a correlation with the patient's neurologic condition. This case illustrates the difficulties in differentiating dysesthesia accompanying cutaneous manifestations of exanthem from neurologic disease on the basis of the clinical appearance alone.

Adolescent↗