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PubMed1997

Characterization of a nuclear localization signal of canine parvovirus capsid proteins.

We investigated the abilities of synthetic peptides mimicking the potential nuclear localization signal of canine parvovirus (CPV) capsid proteins to translocate a carrier protein to the nucleus following microinjection into the cytoplasm of A72 cells. Possible nuclear localization sequences were chosen for synthesis from CPV capsid protein sequences (VP1, VP2) on the basis of the presence of clustered basic residues, which is a common theme in most of the previously identified targeting peptides. Nuclear targeting activity was found within the N-terminal residues 4-13 (PAKRARRGYK) of the VP1 capsid protein. While replacement of Arg10 with glycine did not affect the activity, replacement of Lys6, Arg7, or Arg9 with glycine abolished it. The targeting activity was found to residue in a cluster of basic residues, Lys5, Arg7, and Arg9. Nuclear import was saturated by excess of unlabelled peptide conjugates (showing that it was a receptor-mediated process). Transport into the nucleus was an energy-dependent and temperature-dependent process actively mediated by the nuclear pores and inhibited by wheat germ agglutinin.

Adenosine Triphosphate↗
PubMed2006

Parvovirus B19-induced anemia in renal transplantation: a role for rHuEPO in resistance to classical treatment.

Human parvovirus B19 (PVB 19) is responsible for pure red cell aplasia in immunocompromised patients, and particularly solid organ recipients. Intravenous immunoglobulins (IVIG) have been shown to be efficient to achieve the correction of anemia in association with the reduction of immunosuppression. We report a case of kidney transplant recipient with PVB 19-induced anemia that did not respond to recombinant human erythropoietin (rHuEPO) and to a first course of IVIG. After discontinuation of rHuEPO, a second course of IVIG was successful with the resolution of anemia. We discuss the role of rHuEPO that may facilitate PVB 19 replication in erythropoietin-sensitive human erythroid progenitor cells.

Drug Resistance↗
PubMed1994

Detection of canine parvovirus DNA in paraffin-embedded tissues by polymerase chain reaction.

Canine Parvovirus (CPV) is seemingly a 'new' virus which suddenly appeared during the mid-1970's in an epizootic of disease in dogs. The virus is very similar to the feline panleukopenia virus (FPV), and recent studies have underlined the possible emergence of CPV as a variant of a virus from some other carnivore--possibly from FPV (Parrish, 1990). Several conserved amino-acid changes between CPV and FPV isolates have been defined by cloning and sequencing the capsid-protein gene. An alternative to cloning and sequencing the entire capsid-protein gene would be to use PCR amplification of short regions of the gene containing the appropriate variable amino-acid codons. In addition, use of PCR would also facilitate the study of virus samples which cannot be recovered as infectious agents, e.g. after having undergone formalaldehyde fixation and paraffin-embedding procedures. This study reports on the amplification of CPV DNA from 15-year-old tissue sections which have been prepared by formaldehyde or paraformaldehyde-lysine-periodate-glutaraldehyde fixation, using PCR with various primer pairs within the capsid-protein gene of CPV.

Animals↗
PubMed1995

Differentiation of wild- and vaccine-type canine parvoviruses by PCR and restriction-enzyme analysis.

The polymerase-chain reaction (PCR) and restriction-fragment-length-polymorphism (RFLP) analysis were used to differentiate the wild- and vaccine-type of canine parvovirus (CPV) in Japan. The entire coding region of the CPV genome was enzymatically amplified, and the PCR products of three wild strains and four vaccine strains were analysed using RFLP assay. Then, two polymorphic regions in the VP1/VP2 gene were selected to generate strain-specific RFLP patterns. By using four restriction enzymes, wild and vaccine strains were clearly differentiated; only two vaccine strains, probably of the same origin, were indistinguishable from each other. The wild strains retained strain-specific RFLP patterns throughout in vitro passage, and there was no diversity of RFLP patterns among the different lots of vaccine strains. A total of 21 recent field samples were tested, showing RFLP patterns identical to those of a wild strain isolated in 1991. These results suggest that the PCR-RFLP analysis is a practical and reliable method of differentiating wild- and vaccine-type CPVs.

Animals↗
PubMed1996

Detection and genomic analysis of canine parvovirus by the polymerase chain reaction.

Prevalence of canine parvovirus type 2 (CPV-2) in Japanese dogs and genomic variations among the virus strains were examined. Two-step polymerase chain reaction with double-nested primer pairs designed in the NS and VP1/VP2 genes of CPV-2 was developed for the detection of the viral genome in faecal samples. A total of 74 samples obtained from diarrhoeal house dogs between 1993 and 1995 were tested by the PCR. The virus-positive rate was 54.1%, showing that CPV-2 is still involved in many cases of acute infectious diarrhoea in Japanese dogs. The VP1/VP2 gene of the positive samples was subjected to restriction fragment length polymorphism (RFLP) analysis and nucleotide sequencing. RFLP patterns of the samples were almost identical to those of one CPV-2 strain (TDKet-91-42) isolated in 1991, but different from those of the CPV-2 in the late 1970s and 1980s. The results suggest that a new genotype of CPV-2 appeared and spread among Japanese dogs in the early 1990s.

Animals↗
PubMedDate not supplied

New approaches for the molecular characterization of canine parvovirus type 2 strains.

Characterization of the canine parvovirus type 2 (CPV-2) is sometimes ambiguous, frequently requiring more than one technique for definitive prediction of the viral type. Taking into account the single-nucleotide polymorphisms encountered in the VP2-protein gene between types 2a and 2b and between type 2b and Glu-426 mutant (type 2c), two different minor groove binder (MGB) probe assays were developed for rapid identification of the CPV-2 variants. A total of 315 samples collected from dogs with diarrhoea were screened for CPV-2 by a real-time polymerase chain reaction (PCR) assay capable of detecting all CPV-2 types. In order to compare the type-specific assays with the traditional techniques [haemagglutination inhibition with monoclonal antibodies, PCR-restriction fragment-length polymorphism (RFLP), sequence analysis] for prediction of CPV-2 antigen specificity, the 203 samples tested CPV-2 positive were analysed using the different methods. The results showed a 100% concordance between the MGB probe assays and the combined conventional methods, with 116 samples characterized as type 2a, 32 as type 2b and 55 as type 2c. Therefore, the MGB probe assays represent a quick, reliable tool for prediction of CPV-2 antigen specificity, with regard to the more time-consuming assays currently used.

Animals↗
PubMedDate not supplied

Antibody levels and protection to canine parvovirus type 2.

The relationship between maternally derived antibody (MDA) levels and protection to canine parvovirus (CPV) infection in pups is reported. Twelve pups with a wide range of haemagglutination inhibiting (HI) titres of MDA to CPV were divided into four groups, with each group balanced for antibody titres. The dogs were inoculated with a field CPV-2b strain and clinical signs, virus shedding and antibody response were assessed. The CPV was not detected in the faeces of dogs with HI titres of 320 at any time. In dogs with HI titres up to 160, active CPV replication after challenge was demonstrated by real-time polymerase chain reaction. The successful infection of dogs with HI titres of 80 and 160 was confirmed by seroconversion, evaluated at day 14 post-infection. These findings demonstrated that CPV infection could also occur in the presence of MDA HI titres (> or =80) usually considered fully protective.

Animals↗
PubMed2006

Immune complex-based vaccine for pig protection against parvovirus.

The insoluble immune complexes (ICs) were prepared under the conditions of double immunodiffusion in gel, using the suspension of the ultrasound treated PK-15 cell-line infected with porcine parvovirus (PPV) containing both viral particles and viral proteins, as well as pig or rabbit anti-PPV polyclonal immune sera. The immunodiffusion performed in an agarose gel allows only viral subunits with a molecular mass equal to or less than 1000 kDa, rather than the viral particles, to diffuse through the gel and reach the point where the immunoprecipitate is to be formed. The immunoprecipitation under the conditions of the diffusion ensures the optimal, i.e. equimolar ratio of both immunoprecipitating components, antibody/antigen in the IC. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the Western blot analyses showed the ICs were composed of two proteins, a protein in which molecular mass corresponded to the VP2 of the PPV and a protein with a molecular mass of the IgG. This suggests that the ICs are mainly composed of the VP2 antigen and IgG class antibodies. The potency of the IC-vaccines prepared in the form of a water-in-oil-in-water emulsion was compared with that of a commercially available, inactivated oil vaccine. The vaccination of gilts, 6 weeks before mating, with the IC containing allogeneic pig antibodies, resulted in the development of high and long-lasting anti-PPV antibody titres, similar to those generated by the licenced vaccine (P > 0.01). The content of the virus material administered by the IC was twice lower than that in the licenced vaccine. Neither systemic nor local reactions were observed in the gilts during the period of the trial with the IC vaccine. The number of viable piglets per litter varied between 9 and 12 and no signs of the PPV infection were detected. Rabbits were used as one of the alternative laboratory animal models accepted for the testing of the vaccine against the PPV. The rabbit humoral immune response generated by the IC containing the allogeneic antibodies were higher than that generated by the ICs containing the xenogeneic pig antibodies. It was similar to that generated by two-times higher content of the virus material administered by a commercially available vaccine. The IC-based vaccines belong to non-replicating, subunit vaccines, which are both ecologically convenient and the safest vaccines of all.

Animals↗
PubMed2006

First detection of canine parvovirus type 2c in pups with haemorrhagic enteritis in Spain.

Canine parvovirus type 2 (CPV-2), the aetiological agent of haemorrhagic enteritis in dogs, includes three antigenic variants, types 2a, 2b and 2c. CPV-2c has been detected initially in Italy and subsequently in Vietnam. We report the first identification of this novel antigenic variant in Spain, where it caused an outbreak of fatal enteritis in basset hound pups in association with canine coronavirus type I and type II. We suggest that this new antigenic variant of CPV-2 could spread throughout Europe and that there is a subsequent need to update current CPV vaccines.

Animals↗
PubMed2005

High porcine parvovirus antibodies in sow herds: prevalence and associated factors.

Porcine parvovirus (PPV) is widespread among swine. Our objective was to determine the prevalence of loosely housed sow herds in Finland with at least one animal with high (infection level) PPV antibodies and to gather basic knowledge about vaccination practices. In addition, selected factors associated with high antibody levels found in sows were examined. Altogether, 247 animals were sampled in 21 randomly chosen loosely housed sow herds. Samples were analysed with the haemagglutination inhibition (HI) test. PPV proved to be common; in 17 farms (81%) at least one animal had a high titre (>1 : 512), and 44% of all animals sampled had a high titre. The vaccination programmes had many shortcomings. In the generalised estimation equations (GEE) population-averaged model developed, the factors found to have a significant (p < or = 0.05) effect on HI titres were herd size, parity of two or greater and storage of the vaccine vial after use. Non-returning rate, re-breeding interval and litter size did not differ between herds with no high HI titres (n = 4) and those with at least one high HI titre (n = 17).

Animals↗
PubMed2006

Parvovirus antibodies in vaccinated gilts in field conditions--results with HI and ELISA tests.

This study was conducted to determine the antibody response for porcine parvovirus (PPV) of 39 gilts in field conditions after vaccination. Gilts from four herds endemically infected with PPV were injected twice with a commercial vaccine of inactivated PPV and Erysipelothrix rhusiopathiae. The PPV antibodies were analysed both with haemagglutination inhibition (HI) and enzyme-linked immunosorbent assay (ELISA) in order to study the agreement between these methods. The possible association between high-antibody titres and reproductive failure (repeat breeding, culling for infertility, < or = 6 piglets born alive) was also investigated. In these study herds, endemically infected by PPV, most gilts (84.6%) had not seroconverted by the age of 6 months. On-field vaccination resulted in a consistent increase of humoral immunity not exceeding the antibody level of 1 : 512 in the majority of gilts in all herds examined. The agreement between ELISA and HI tests was moderate (Spearman's rho = 0.87, kappa = 0.63). The seroconversion over the level >1:512 by mid-pregnancy was not associated with reproductive failure.

Agglutination Tests↗
PubMed1998

Parvovirus B19-associated transient pure red cell aplasia with lymphadenopathy: a case report.

There have been few reports on lymph node swelling in human parvovirus (HPV) B19 infection. A report of a 42-year-old female, who developed HPV B19-associated transient red cell aplasia with lymphadenopathy, is presented. The lymph node swelling began with the appearance of atypical lymphocytes in the peripheral blood and it disappeared as the patient recovered from the aplasia. Microscopically, the patient's bone marrow showed characteristic giant proerythroblasts with no maturation of the erythroid series. An excised inguinal lymph node showed florid, reactive follicular hyperplasia with paracortex expansion, and neutrophil infiltration and hemophagocytosis in the medullary sinus. These findings were compatible with the histology of a viral infection. A polymerase chain reaction study revealed HPV B19 in her serum and lymph node, but an immunohistochemical study failed to demonstrate HPV B19 capsid antigen in the lymph node or bone marrow. Although the present case suggests that reactive lymphadenopathy is associated with HPV B19 infection, the mechanism of the lymph node swelling still remains to be elucidated.

Adult↗
PubMed1994

Transient erythroblastopenia due to human parvovirus B19 infection: a case report of a boy suffering from purpura.

Transient erythroblastopenia of childhood (TEC) was observed in a boy infected by human parvovirus B19 (B19) before the manifestation of the typical clinical picture of erythema infectiosum (EI). He did not have any apparent symptoms of anemia, and spontaneous resolution of erythroblastopenia was observed before the manifestation of the symptoms of EI. The difference between primary B19 infection and classical TEC is discussed.

Child↗
PubMed1994

Human parvovirus B19-induced aplastic crisis in iron deficiency anemia.

Human parvovirus B19 (HPVB19) infects and replicates in erythroid progenitor cells. Its specific cytotoxic effect on these cells results in aplastic crises in patients with congenital hemolytic anemias. Aplastic crisis due to HPVB19 infection in a healthy girl revealed occult iron deficiency anemia. The condition is characterized by a high serum iron level in the aplastic phase and rapid recovery after administration of iron. Temporary HPVB19-induced red blood cell aplasia could occur in patients with other anemias, particularly those with non-inherited form of hemolysis.

Adolescent↗
PubMed1996

Leukophagocytosis in human parvovirus B19-induced transient bicytopenia in a healthy child.

A previously health 11 year old boy had transient leukopenia and thrombocytopenia associated with human parvovirus B19 (B19) infection. This case suggested that low-grade hemophagocytosis can be involved in developing B19-related cytopenia even in an otherwise healthy child. Bone marrow aspiration at an early stage could reveal the underlying mechanism of B19-related cytopenia.

Biopsy, Needle↗
PubMed1996

A case of prolonged human parvovirus B19 DNA-emia associated with polyclonal B cell activation.

The current paper reports an 8 year old girl with arthralgia and polyclonal B cell activation induced by human parvovirus B19 infection (HPV B19). The infection was diagnosed by the presence of the virus genome in sera. The patient presented with transient arthritis in the wrist, ankle joint and neck and elevation of immunoglobulin IgM antibodies to HPV B19 and rubella, antibodies to Mycoplasma and antistreptolysin O but without the typical clinical features of erythema infectiosum. The polyclonal B cell activation was paralleled by the presence of the virus genome of HPV B19 in sera. In some children with arthralgia, it is important to examine the genomes of viruses that may cause arthritis as well as the antibody titers to the viruses.

Arthralgia↗
PubMed1998

Human parvovirus B19-associated thrombocytopenic purpura.

We report two patients who presented with thrombocytopenic purpura (TP) associated with primary human parvovirus B19 (B19) infection. One patient also had transient liver dysfunction. In both cases, B19-DNA was detected in serum and bone marrow by polymerase chain reaction. Six months after the illness in patient 1 and 8 months after the illness in patient 2, B19-DNA disappeared from the serum. Serum immunoglobulin (Ig)G antibody to B19 remained positive in both cases, but B19 IgM antibody became negative 3 months after the onset in case 1 and 4 months after the onset in case 2. The mechanism of TP by B19 infection is unknown.

Adolescent↗