Estimation of serum concentration of parvovirus B19 DNA by PCR in patients with chronic anaemia

Hornsleth, A; Carlsen, K. M.; Christensen, Laurids Siig; Gundestrup, Maryanne; Heegaard, Erik D.; Myhre, J.

doi:10.1016/s0923-2516(07)80043-4

Cited by 26 publications

(20 citation statements)

References 14 publications

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“…As part of a retrospective pilot study of parvovirus B19 infection in BM recipients, we tested a number of stored BM and serum samples from the patient by enzyme-linked immunosorbent assay (ELISA) and amplified the NS and VP genes by nested polymerase chain reaction (PCR) (Hornsleth et al, 1994). Examination of this material revealed a prolonged parvovirus B19 infection, as evidenced by the finding of parvovirus B19-specific DNA and antibodies (Table I).…”

Section: Patient and Methodsmentioning

confidence: 99%

“…Transfusion of 16 units of red blood cells and 101 units of platelets were needed to sustain and correct these parameters. The patient then finished the standard protocol involving methotrexate and cyclosporin treatment and has had no adverse haematological symptoms during 15 years of follow-up.As part of a retrospective pilot study of parvovirus B19 infection in BM recipients, we tested a number of stored BM and serum samples from the patient by enzyme-linked immunosorbent assay (ELISA) and amplified the NS and VP genes by nested polymerase chain reaction (PCR) (Hornsleth et al, 1994). Examination of this material revealed a prolonged parvovirus B19 infection, as evidenced by the finding of parvovirus B19-specific DNA and antibodies (Table I).…”

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confidence: 99%

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Parvovirus B19 transmitted by bone marrow

Heegaard¹,

Petersen

2000

Br J Haematol

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Summary. We describe a case of symptomatic parvovirus B19 infection transmitted by bone marrow (BM). The infection caused prolonged anaemia, thrombocytopenia, arthralgia and erythema infectiosum in a 16-year-old girl with acute myeloid leukaemia receiving a BM transplant (BMT). The BM donor was a 19-year-old asymptomatic brother who had parvovirus B19 viraemia at the time of BM harvest. Sequencing of the VP2 gene from the patient and the donor showed a perfect match of DNA sequences, confirming the mode of transmission. Parvovirus B19 represents a potential complicating factor in patients undergoing BMT, but screening by polymerase chain reaction (PCR) of donor BM may reduce the risk of infection.Keywords: bone marrow, ELISA, parvovirus B19, polymerase chain reaction, transplantation.Parvovirus B19 exhibits a marked tropism to human bone marrow (BM) and replicates only in erythroid progenitor cells (Brown & Young, 1995). The virus is transmissible by the respiratory route or by blood and infection may lead to erythema infectiosum, arthropathy, hydrops fetalis and various haematological disorders (aplastic crisis, chronic anaemia, idiopathic thrombocytopenic purpura). We present the first case of symptomatic parvovirus B19 infection transmitted by BM causing prolonged anaemia, thrombocytopenia, arthralgia and erythema infectiosum. PATIENT AND METHODSA 16-year-old girl diagnosed with acute myeloid leukaemia underwent bone marrow transplantation (BMT) on 25 October 1984 (day 0). Twelve days later, she developed fever and a rash involving the upper extremities. The rash subsequently progressed, involving the lower extremities, neck and face (slapped cheek appearance). Coinciding with these findings, the patient complained of arthralgia, mainly involving the large joints. The symptoms gradually disappeared over a 3-week period.Examination of BM slides (air-dried BM inoculum) using light microscopy revealed no lantern cells or giant pronormoblasts at the time of BMT or during follow-up. Apart from red cell hypoplasia, no specific changes were noted. Reticulocytes were absent for extended periods of time (days 0±15 and 35±75) and reoccurred briefly in small numbers (days 16±34), before showing a slow but steady increase towards normal levels (day 76 onwards). Within the first 76 d, haemoglobin levels were accordingly low (5´6±11´6 g/dl), and a reduced number of platelets (5± 78 Â 10 9 /l) were observed. Transfusion of 16 units of red blood cells and 101 units of platelets were needed to sustain and correct these parameters. The patient then finished the standard protocol involving methotrexate and cyclosporin treatment and has had no adverse haematological symptoms during 15 years of follow-up.As part of a retrospective pilot study of parvovirus B19 infection in BM recipients, we tested a number of stored BM and serum samples from the patient by enzyme-linked immunosorbent assay (ELISA) and amplified the NS and VP genes by nested polymerase chain reaction (PCR) (Hornsleth et al, 1994). Examination of this material reve...

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Section: Patient and Methodsmentioning

confidence: 99%

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confidence: 99%

Parvovirus B19 transmitted by bone marrow

Heegaard¹,

Petersen

2000

Br J Haematol

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“…This can explain the imprecise quantification and the high interlaboratory variability of detection. Nevertheless, the high frequency of myocardial B19V genome detected both in IDC and normal LVEF by means of PCR, immediately raises the question of a possible contamination leading to false positive results in this extremely specific and sensitive PCR method as discussed previously [Hornsleth et al, 1994]. However, nucleotide variability in the VP1 region of the parvoviral genome was shown by sequence analysis of all B19V-positive endomyocardial biopsies from the IDC patients.…”

Section: Methodological Aspectsmentioning

confidence: 82%

Low level myocardial parvovirus B19 persistence is a frequent finding in patients with heart disease but unrelated to ongoing myocardial injury

Lotze¹,

Egerer

Glück

et al. 2010

Journal of Medical Virology

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While myocardial parvovirus B19 (B19V), aside from enteroviruses (EV) and adenoviruses (ADV), has recently been found often in patients with myocarditis and idiopathic dilated cardiomyopathy (IDC), the pathogenetic significance of B19V genomes in those patients has not yet been sufficiently elucidated. In the present study, left ventricular endomyocardial biopsies from 24 patients with left ventricular ejection fraction (LVEF) below 55% due to IDC, and tissue from the right atrial appendage of 10 control patients undergoing bypass surgery with normal LVEF (>55%) were investigated for B19V, ADV, and EV genomes by specific nested polymerase chain reaction (PCR), by real time PCR or by reverse‐transcription PCR, respectively. The myocardial tissue samples from the 10 controls were analyzed each in three different virological laboratories for B19V. In the IDC group, the frequency of the myocardial virus genomes found in 54% (13/24) of the patients was as follows: B19V: 50% (12/24), EV: 8% (2/24), including one patient with B19V and EV, and ADV: 0% (0/24). For comparison, the prevalence of B19V genomes was between 30% and 60% in the control group as detected in three different laboratories, but all these control subjects were EV‐ and ADV‐negative. The number of B19V gene copies, however, was very low and similar both in the IDC and control group. In the majority of patients myocardial B19V persistence was associated with a low virus load irrespective of the underlying heart disease so that it may be of no importance in the pathogenesis of IDC. J. Med. Virol. 82:1449–1457, 2010. © 2010 Wiley‐Liss, Inc.

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“…Despite these discrepancies, the kappa coefficient revealed a very good correlation between the LC-FRET PCR and the block cycler PCR formats. The sensitivity of the LC-FRET PCR is in the range of other published B19 DNA quantification methods, i.e., approximately 5 geq per assay (1,4,10,17,22,25,40). For immunocompetent patients, LC-FRET PCR also correlated favorably with IgM serostatus ( ϭ 0.92).…”

Section: Discussionmentioning

confidence: 95%

“…Detection of DNA by hybridization or PCR has been reported to be superior in the diagnosis of prenatal B19 infections and in children with oncologic or hematologic disorders (5,11,34). Several qualitative and quantitative PCR methods targeting different regions of the parvovirus genome have been published (1,4,9,10,13,15,17,22,25,40). These PCR approaches are based on conventional block thermocycling and combined single-round PCRs with subsequent oligohybridization or use a nested format.…”

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New LightCycler PCR for Rapid and Sensitive Quantification of Parvovirus B19 DNA Guides Therapeutic Decision-Making in Relapsing Infections

Harder¹,

Hufnagel

Zahn³

et al. 2001

J Clin Microbiol

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The human erythrovirus B19, a member of the family Parvoviridae, causes a broad and seemingly expanding spectrum of disorders (7,12,23,44). The clinical picture depends on the immune status and age of the patient (14,36,43). Parvovirus B19 shows a remarkable tropism for erythroid progenitor cells in the bone marrow, which is partly based on binding to its receptor, the blood group P antigen (6). Viral replication, and possibly also the induced immune response, interferes with physiological functions and loss of the erythroid progenitor cells, resulting in a usually subclinical reticulocytopenia (8). In patients with an underlying hematologic disorder and high blood cell turnover (e.g., hemolytic anemia), transient aplastic anemia may ensue. Immunocompromised patients, who have an increased risk of developing a persistent B19 infection, are threatened by a chronic reticulocytopenic anemia, also known as pure red cell anemia (2,8).Diagnosis of uncomplicated cases of acute B19 infection (fifth disease or arthropathy) is usually clinically based and can be accomplished by detection of specific immunoglobulin M (IgM) antibodies except in immunocompromised patients, who are prone to persistent infection and who may generate IgMspecific B19 antibodies less reliably (7,19). Likewise, specific IgG is not a reliable marker for discriminating a reconvalescent status from chronic persistent infection (35), although recent data indicate that IgG antibodies specific for nonstructural protein 1 (NS-1) of B19 are more frequently associated with persistent infection (24). Detection of DNA by hybridization or PCR has been reported to be superior in the diagnosis of prenatal B19 infections and in children with oncologic or hematologic disorders (5,11,34). Several qualitative and quantitative PCR methods targeting different regions of the parvovirus genome have been published (1,4,9,10,13,15,17,22,25,40). These PCR approaches are based on conventional block thermocycling and combined single-round PCRs with subsequent oligohybridization or use a nested format.The recently developed LightCycler (LC) DNA amplification technology (Roche Diagnostics, Mannheim, Germany) combines rapid glass capillary thermal cycling with real-time microvolume fluorescence monitoring (47). Detection of amplicons is achieved during the run in real time. Melting point analysis of the amplicons at the end of the run is used as a specificity control when the fluorochrome SYBR green is used for detection of double-stranded DNA. Alternatively, specificity can be tested during the run by using two target-specific hybridization probes which utilize fluorescence resonance energy transfer (FRET) to generate a measurable signal. In the latter case, two oligonucleotide probes (HybProbe) bind to immediately adjacent regions of the respective amplicon. The upstream probe (referred to as the probe) is labeled at its 3Ј end with fluorescein, while the 5Ј end of the downstream probe (referred to as the anchor) is labeled with either of the fluorochromes LC-Red 640 and LC-...

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Estimation of serum concentration of parvovirus B19 DNA by PCR in patients with chronic anaemia

Cited by 26 publications

References 14 publications

Parvovirus B19 transmitted by bone marrow

Parvovirus B19 transmitted by bone marrow

Low level myocardial parvovirus B19 persistence is a frequent finding in patients with heart disease but unrelated to ongoing myocardial injury

New LightCycler PCR for Rapid and Sensitive Quantification of Parvovirus B19 DNA Guides Therapeutic Decision-Making in Relapsing Infections

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