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Persistent parvovirus B19 infections in immunocompromised children.

Immunocompromised patients have been shown to suffer from prolonged viral infections often without detectable immune response. Here chronic infections with low virus levels can be frequently observed. In these patients viral DNA can be detected over long periods by polymerase chain reaction (PCR). In this study parvovirus B19 presence was assessed by PCR, immunoblot and enzyme-linked immunosorbent assay in sera from children with mainly oncological and hematological diseases. In 45% of sera B19 DNA was observed. Of the children 25% had IgG antibodies to viral protein 1 and 2 (VP1/2) and 15% to nonstructural protein 1 (NS1). In 6% of children IgM antibodies to VP1/2 were detected. These results indicate that the number of children with immune response to B19 proteins is distinctly lower than the number of children with B19 DNA. Transfusions of blood products might have been a possible route for B19 infection. Establishment and maintenance of a persistent parvovirus B19 infection with or without immune response are enhanced in the analyzed immunocompromised children in comparison with immunocompetent children. A persistence of B19 DNA was demonstrated up to 10 months in patients sera.

Adolescent↗

Analysis of VP2 gene sequences of canine parvovirus isolates in India.

The epidemiology of canine parvovirus (CPV) infections in dogs in India was examined using 27 isolates collected during a two-year period. The VP2 genes of 22 isolates were sequenced, and the deduced amino acid sequences were compared. The results indicated that the isolates belonged to CPV type 2a except four, which belonged to CPV type 2b. Comparison of the VP2 gene sequences revealed that the Indian isolates formed separate lineages distinct from the South East Asian isolates. The canine parvovirus isolates in India appear to evolve independently, and distinct geographical patterns of evolution could not be discerned in the isolates examined.

Animals↗

Multiple small pulmonary emboli associated with transient antiphospholipid syndrome in human Parvovirus B19 infection.

Antiphospholipid antibodies (aPL) have been reported to occur in several conditions other than antiphospholipid syndrome, including infections. We herein report the case of a 21-year-old Japanese woman with Parvovirus B19 infection, who developed multiple pulmonary emboli associated with aPL, a lupus anticoagulant and IgM anticardiolipin antibody. Eight weeks later, antiphospholipid antibodies spontaneously disappeared and normal pulmonary flow was observed. Considering the high prevalence of Parvovirus B19 infection, we should be aware of thrombosis associated with transient aPL antibodies in this infectious disease.

Adult↗

Occurrence and clinical role of active parvovirus B19 infection in transplant recipients.

To evaluate the occurrence and clinical role of active parvovirus B19 infection in solid organ and bone marrow transplant recipients, 256 serum samples from 212 transplant patients were investigated retrospectively by competitive polymerase chain reaction. Sera were drawn during the transplantation period and up to 6 months after transplantation during a nonepidemic 1-year period. Three patients were found positive for B19 DNA; only one liver transplant patient had a clinically overt B19 infection. Overall, the rate of active parvovirus B19 infection in transplant subjects was low (1.42%), probably due to the high number of actively or passively immunized subjects among transplant recipients; this may also account for the asymptomatic infections observed.

Adult↗

Genomic typing of canine parvovirus circulating in the State of Rio de Janeiro, Brazil from 1995 to 2001 using polymerase chain reaction assay.

In this study, the genomic types of canine parvovirus (CPV) circulating in the State of Rio de Janeiro, Brazil, from 1995 to 2001, were investigated using the polymerase chain reaction assay (PCR). A total of 78 faecal samples from gastroenteritic puppies, confirmed as positive for canine parvovirus by haemagglutination/haemagglutination inhibition tests or virus isolation in cell culture (MDCK), were examined. The viral DNA was extracted from faecal samples using a combination of phenol- chloroform and silica-guanidine thiocyanate methods. PCR was carried out with differential pairs of primers to distinguish the old (CPV-2) and new types of virus (CPv-2a or CPV-2b). Specific amplicons were observed for all samples using the primer pair P2ab, which detects CPV-2a and CPV-2b. Seventy-six from a total of 78 samples (97%) were considered as CPV-2b because of their reaction with the primer pair P2b. Thirty samples (30/78) were from previously vaccinated puppies and in 15 of them the enteritis symptoms began from 1 to 12 days after vaccination. PCR confirmed the infection by wild virus (CPV-2b) in 5 of these 15 puppies who had received old-type vaccines. Our results show that CPV-2b was the prevalent type circulating in the State of Rio de Janeiro from 1995 to 2001.

Animals↗

Sequence analysis of a canine parvovirus isolated from a red panda (Ailurus fulgens) in China.

Canine parvovirus (CPV) was first recognized in the late 1970 s in dogs and has mutated and spread throughout the world in canid and felid species since then. In this study, a novel CPV was isolated from the endangered red panda (Ailurus fulgens) in China. Nucleotide and phylogenetic analysis of the capsid protein VP2 gene classified the red panda parvovirus (RPPV) as a CPV-2a type. Substitution of Val for Gly at the conserved 300 residue in RPPV presents an unusual variation in the CPV-2a amino acid sequence and is further evidence for the continuing evolution of the virus. The 300 residue is important in distinguishing the antigenicity and host range of CPVs. The clinical significance and population impact of RPPV infection in captive red pandas in China is unknown and is an important topic for future research.

Ailuridae↗

Cytokines in parvovirus B19 infection as an aid to understanding chronic fatigue syndrome.

Human parvovirus B19 infection has been associated with various clinical manifestations of a rheumatic nature such as arthritis, fatigue, and chronic fatigue syndrome (CFS), which can persist for years after the acute phase. The authors have demonstrated recently that acute B19 infection is accompanied by raised circulating levels of IL-1b, IL-6, TNF-a, and IFN-g and that raised circulating levels of TNF-a and IFN-g persist and are accompanied by MCP-1 in those patients who develop CFS. A resolution of clinical symptoms and cytokine dysregulation after intravenous immunoglobulin (IVIG) therapy, which is the only specific treatment for parvovirus B19 infection, also has been reported. Although CFS may be caused by various microbial and other triggers, that triggered by B19 virus is clinically indistinguishable from idiopathic CFS and exhibits similar cytokine abnormalities and may represent an accessible model for the study of CFS.

Chemokine CCL2↗

Structure of the 3' hairpin termini of four rodent parvovirus genomes: nucleotide sequence homology at origins of DNA replication.

The nucleotide sequences of the 3' termini of the DNA from four autonomous rodent parvoviruses have been determined. The terminus of each genome exists as a Y-shaped hairpin structure involving 115 or 116 nucleotides. The sequence of this region of DNA is highly conserved and shows no evidence of internal sequence heterogeneity, a characteristic which is observed in the terminal nucleotide sequence of the helper-dependent, adeno-associated viruses (Berns et al., 1978a). The implications of these results with respect to the models of parvovirus DNA replication are discussed.

Base Sequence↗

Expression of the autonomous parvovirus H1 genome: evidence for a single transcriptional unit and multiple spliced polyadenylated transcripts.

We identified viral transcripts in parvovirus H1-infected rodent cells using the S1 nuclease mapping technique of Berk and Sharp (1977, 1978). The most abundant viral transcript, present in both nucleus and cytoplasm, is approximately 2.8 kb long and represents about 56% of the viral genome. Less abundant viral transcripts of 3.0, 1.45 and 1.30 kb, and possibly other minor viral transcripts, are also detected in nuclear and cytoplasmic fractions. In contrast, a prominent 4.7 kb viral transcript which corresponds to 95% of the viral DNA is found only in the nucleus; this finding suggests that the parvovirus genome may function as a single transcription unit. Virus-infected cells pretreated with cycloheximide accumulate all these viral transcripts. Analyses of RNA-DNA hybrids (isolated from neutral agarose gels) by electrophoresis on alkaline agarose gels indicate that the 4.7, 3.0 and 2.8 kb viral transcripts are "spliced" RNAs. The nuclear-specific 4.7 kb transcript appears to be encoded by two noncontiguous DNA segments of 2.2 and 2.6 kb. The 3.0 and 2.8 kb transcripts are apparently encoded by.a 2.6 kb segment of DNA and one or more much smaller noncontiguous DNA segments, one of which is approximately 170 nucleotides long.

Animals↗

Parvovirus B19 outbreak in a children's ward.

Parvovirus B19 infection can cause severe complications in pregnant women, individuals with haemolytic anaemia, and those who are immunocompromised. In a hospital outbreak of this infection, a balance should be struck between protection of these individuals and the maintenance of medical services. The index case of an outbreak of parvovirus B19 infection among staff and patients of a paediatric ward was not identified. 58 members of staff were screened for B19 markers and 4 of the 6 susceptible men and 6 of the 24 susceptible women became infected (p = 0.05) as defined by serum IgM and viraemia. 1 of the 11 adults (10 members of staff and 1 parent) infected remained symptom-free. 12 immunocompromised patients were also assessed, and symptom-free infection developed in 2 of these. During the outbreak staff with symptoms were put on sick leave, immunocompromised patients (there were none with haemolytic anaemia) were given normal human immunoglobulin and nursed in single rooms by B19 IgG-positive, IgM-negative staff, and the ward was closed to B19 IgG-negative pregnant women. However, the limitation of spread of infection cannot be attributed with certainty to the measures taken.

Adult↗

IgG and IgM antibodies to human parvovirus B19 in the serum of patients with a clinical diagnosis of infection with the virus and in the general population of Saudi Arabia.

A total of 56 samples of serum from 32 patients with a clinical diagnosis of human parvovirus B19 infection were tested for specific immunoglobulin G (IgG) and M (IgM) antibodies by means of the recently available indirect enzyme-linked immunosorbent assay (ELISA) (Parvoscan-B19, Ferring Diagnostica, Sweden). The assay was also used in order to determine the age-specific prevalence of antibodies to the virus in the general population of Saudi Arabia. Specific IgM antibodies were detected in 94% specimens collected 1 week after the onset of illness and could be detected for up to 2 months. On the other hand, specific IgG antibodies were detected in 85% patients from whom acute- and convalescent-phase serum samples were collected. Saudis begin to be exposed to human parvovirus B19 early in life and prevalence of exposure increases with age in both sexes (overall prevalence 19.0%). The availability of a commercial ELISA makes it possible to diagnose infection with the virus routinely and will help in establishing the extent of exposure to it in various communities.

Adolescent↗

Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19.

A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.

Base Sequence↗

In situ hybridisation and in situ polymerase chain reaction detection of parvovirus B19 DNA within cells.

Modification of an in situ polymerase chain reaction (ISPCR) technique is described for the detection of B19 parvovirus infection. Specific amplification of B19 DNA inside fixed cells was followed by hybridisation with a digoxigenin-labelled probe and then visualised by immunochemical reaction. The assay had higher sensitivity compared to direct in situ hybridisation and still allowed cellular localisation and characterisation of infected cells. This assay can be used as a confirmatory method for PCR in tissues and will allow further identification of tissues permissive for B19 parvovirus infection.

Base Sequence↗

Chemiluminescent microwell hybridization assay for direct detection of human parvovirus B19 DNA.

A method for the direct detection of human parvovirus DNA in serum samples that uses a digoxigenin-labeled RNA probe to hybridize with target B19 DNA, followed by capture of the hybrid onto a microtiter plate wells previously coated with a second oligonucleotide probe was developed. The captured hybrid is then detected with anti-digoxigenin-alkaline phosphatase conjugate and chemiluminescent substrate and the reaction read on a scintillation counter. The relative sensitivities of the microwell and standard dot blot hybridization assays were compared. The chemiluminescent microwell hybridization assay was more sensitive than dot-blot hybridization and could be performed in a few hours. This format, therefore, permits rapid and sensitive detection of parvovirus DNA suitable for the clinical setting.

Base Sequence↗

Parvovirus B19 outbreak in a developmental center.

Of 49 employees serving developmentally disabled persons, 13 (27%) were found to have parvovirus B19 infection on the basis of serum positivity for parvovirus B19 IgM by enzyme-linked immunosorbent assay. Two employees had severe peripheral symmetric polyarthropathy lasting longer than 6 months. Infection control measures are described.

Adult↗

Effective induction of neutralizing antibodies with the amino terminus of VP2 of canine parvovirus as a synthetic peptide.

Fourteen synthetic peptides corresponding to previously mapped antigenic sites in VP2 of canine parvovirus (CPV) were used for immunization of rabbits to identify antiviral properties favourable for inclusion into a vaccine. Most antipeptide antisera obtained were reactive with viral protein, and with one of them it was possible to locate the hypothetical amino terminus of VP3 within positions 15-31 of VP2. Virus-neutralizing antibodies were only obtained with two overlapping 15-mer peptides corresponding in sequence to the amino terminus of VP2 (MSDGAVQPDGGQPAVRNERAT). Antibodies in the neutralizing sera bound most strongly to amino acids of the sequence DGGQPAV within the N-terminus of VP2, indicating that efforts to develop a synthetic vaccine against CVP should be focused on this stretch of amino acids. The two peptides induced long-lasting immunity (at least 8 months) using either Freund's adjuvant or aluminium hydroxide plus Quil A. Thus, this approach delineated the exact peptide sequence useful for vaccines applied to the amino-terminal region of VP2. These findings in experimental animals form a solid basis for exploration of a synthetic peptide vaccine against parvovirus infection in dogs, minks or cats.

Adjuvants, Immunologic↗

Full protection in mink against mink enteritis virus with new generation canine parvovirus vaccines based on synthetic peptide or recombinant protein.

Two recently developed vaccine--one based on synthetic peptide and one based on recombinant capsid protein--fully protected dogs against heavy experimental canine parvovirus (CPV) infection. The high sequence homology ( > 98%) and antigenic similarity between CPV and mink enteritis virus (MEV), feline panleukopenia virus, and raccoon parvovirus, suggest that both vaccines could protect mink, cats and raccoons against these respective host range variants. This was tested in mink and turned out to be the case. The two vaccines were fully protective and as effective as a conventional commercial vaccine based on inactivated virus. Surprisingly, this protection was obtained after only a single injection. Furthermore, the vaccinal dose of 150 micrograms of conjugated peptide or 3 micrograms of recombinant VP2 particles per animal, are sufficiently low to be cost-effective and applicable on a large scale.

Amino Acid Sequence↗

Human parvovirus B-19: not just a pediatric problem.

Parvoviruses have long been associated with disabling and even fatal illnesses in animals. The discovery of the human parvovirus B-19 in 1975 (1) and subsequent studies of its effects in humans identified this virus as the causative agent of erythema infectiosum ("fifth disease") in children. (2). Erythema infectiosum (EI) is a common, self-limited infectious disorder in children, easily recognized by the classic "slapped cheek" facial erythema and fine reticular rash. Only in the 1980s have further investigations linked HPV B-19 infection with more significant clinical syndromes, among which is an adult polyarthropathy. This presentation in adults is more common than is currently understood and is easily confused with other symmetric polyarthropathies. Recognition and conservative treatment of this disorder are important for the emergency physician, to whom these patients may present.

Adult↗