Novel PCR assay for differential detection and screening of erythrovirus B19 and erythrovirus V9

Heegaard, Erik D.; Jensen, Inge Panum; Christensen, Jesper

doi:10.1002/jmv.2042

Cited by 52 publications

(59 citation statements)

References 10 publications

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“…The geographical distribution of human erythrovirus genotypes is still largely unknown. The predominance of human erythrovirus genotype 1 in the United Kingdom is consistent with previous data; only a few genotype 2 and 3 variants have so far been identified in large-scale screening studies performed in Western countries (15,19,30,35). Until now, genotype 3 strains have occasionally been identified in France, where they have circulated as minority strains accounting for approximately 11% of the strains identified there between 1999 and 2001 (35).…”

Section: Discussionsupporting

confidence: 90%

“…The prevalences observed were significantly higher than the 0.003 to 0.16% prevalences reported in previous studies (2,15,20,27,37,40,41). This discrepancy was clearly unrelated to sample origin but rather reflected differences in the sensitivity of the assays used in the different studies.…”

Section: Discussioncontrasting

confidence: 66%

“…Commercial NAT assays were developed to detect and quantify human erythrovirus genotype 1 or parvovirus B19 and did not take into account genotypes 2 and 3. The reliability of PCR assays designed for the specific detection of genotypes 1 and 3 (15) should also be reconsidered according to recent evidence showing sequence variations in the primer regions (18,35). The data presented in Table 1 indicate that erythrovirus DNA is present at low levels in the circulation of considerably more individuals than previously thought, despite the presence of specific IgG.…”

Section: Discussionmentioning

confidence: 99%

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Identification and Characterization of Persistent Human Erythrovirus Infection in Blood Donor Samples

Candotti

Etiz²,

Parsyan

et al. 2004

J Virol

166

182

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The presence of human erythrovirus DNA in 2,440 blood donations from the United Kingdom and subSaharan Africa (Ghana, Malawi, and South Africa) was screened. Sensitive qualitative and real-time quantitative PCR assays revealed a higher prevalence of persistent infection with the simultaneous presence of immunoglobulin G (IgG) and viral DNA (0.55 to 1.3%) than previously reported. This condition was characterized by a low viral load (median, 558 IU/ml; range, 42 to 135,000 IU/ml), antibody-complexed virus, free specific IgG, and potentially infectious free virus. Human erythrovirus genotype 1 (formerly parvovirus B19) was prevalent in the United Kingdom, Malawi, and South Africa. In contrast, only human erythrovirus genotype 3 (erythrovirus variant V9) was prevalent in Ghana. Genotype 3 had considerable genetic diversity, clustering in two probable subtypes. Genotype 1-based antibody assays failed to detect 38.5% of Ghanaian samples containing antibodies to genotype 3 virus but did not fail to detect cases of persistent infection. This study indicates a potential African origin of genotype 3 human erythrovirus and considerable shortcomings in the tools currently used to diagnose erythrovirus infection. Human erythrovirus (formerly parvovirus B19) is a member of the genus Erythrovirus within the family Parvoviridae (24).Erythrovirus infection occurs frequently in humans. The prevalence of specific immunoglobulin G (IgG) antibodies is 2 to 15% in young children, 30 to 60% in adults, and more than 85% in those 70 years old or older (14). The linear singlestranded DNA genome of this small, nonenveloped virus is about 5 kb long and contains two large open reading frames (ORFs). The first ORF encodes nonstructural protein NS1, and the second one encodes both major VP2 and minor VP1 structural capsid proteins. VP1 consists of a unique sequence of 227 amino acids (VP1u) and is followed by the entire VP2 sequence (554 amino acids). Two additional ORFs encoding small proteins (7.5 and 11 kDa) with unknown functions have also been described (see reference 14 for a review).Following viral infection in immunocompetent individuals, the predominant immune response is a humoral response, which is assumed to confer protective, lifelong immunity. The early IgM response is directed against VP2, while the mature response mostly involves the production of IgG to VP1 (see reference 14 for a review). Several VP2 and VP1u regions containing neutralizing epitopes have been identified (10, 32, 43). However, neutralizing linear epitopes seem to cluster in the N terminus of VP1u and the VP1-VP2 junction regions and to elicit a more efficient response than the epitopes in the VP2 region, which are mainly conformational epitopes (21,26,28,31,36). The inability to develop an efficient neutralizing immune response, as observed mainly in immunosuppressed individuals but also in otherwise healthy individuals, may result in the failure to eliminate the virus, thus leading to persistent infection and the possible occurrence of chronic diseases, suc...

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Section: Discussionsupporting

confidence: 90%

Section: Discussioncontrasting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

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Identification and Characterization of Persistent Human Erythrovirus Infection in Blood Donor Samples

Candotti

Etiz²,

Parsyan

et al. 2004

J Virol

166

182

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“…This method consisted of consensus primers for amplification of both strains, and two pairs of nested primers: one specific for the prototype and the other specific for the variant. Using this method 100 sera containing B19 immunoglobulin M and 50 blood-donor plasma pools were screened without detection of V9 (11). The same result came from 190 bone marrow samples of healthy subjects (10).…”

Section: Discussionmentioning

confidence: 99%

Detection and Differentiation of Human Parvovirus Variants by Commercial Quantitative Real-Time PCR Tests

Hokynar¹,

Norja²,

Laitinen

et al. 2004

J Clin Microbiol

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Parvovirus B19 causes a variety of diseases in humans, with outcomes ranging from asymptomatic to severe, such as chronic anemia in immunocompromised patients or fetal hydrops and death after maternal infection during pregnancy. The virus may be transmitted via plasma-derived products. According to the results of solventdetergent safety studies, an upper limit of B19 DNA in plasma pools was recently defined. To restrict the input of B19 virus into production pools, a quantitative nucleic acid test is a prerequisite. We examined the suitability of the two commercial quantitative B19 PCR tests, LightCycler-Parvovirus B19 quantification kit (Roche Diagnostics) and RealArt Parvo B19 LC PCR (Artus) for detection, quantification, and differentiation of the three known B19 genotypes, including the newly described erythrovirus variants (genotypes 2 and 3). The former kit was highly sensitive for genotype 1 but was not suitable for detection of genotype 2 or one of two genotype 3 strains. The latter kit detected and differentiated all three genotypes, albeit with lower sensitivity for one of the genotype-3 strains. We furthermore assessed the prevalence of the three B19 virus genotypes in blood donors, by screening pooled plasma samples derived from 140,160 Finnish blood-donor units. None of the pools contained detectable levels of B19 virus genotypes 2 or 3. The origin, mode of transmission, and clinical significance of these genotypes are unknown and deserve further study. The RealArt Parvo B19 LC PCR is suitable for detection, quantification, and differentiation of all three B19 virus genotypes in molecular and clinical research.

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“…There was no significant difference for B19 positivity between patients who received blood transfusion and who did not (p>0.05). Additionally, there are several studies which have reported values of 1/167-1/35,000 of B19 positivity in blood donors [18][19][20]. Although B19 infection is known to be transmitted via transfusion, it appears to be unrelated with transfusion; so studies including larger populations are needed to be performed as our study population was relatively small.…”

Section: Discussionmentioning

confidence: 95%

The investigation of parvovirus B19 infection in patients with haematological disorders by using PCR and ELISA techniques

Us¹,

Ozune²,

Kaşifoğlu³

et al. 2007

Braz J Infect Dis

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Parvovirus B19 has a marked tropism for erythroid progenitor cells. This may lead to chronic anemia in predisposed individuals. The purpose of the study was to investigate the frequency of parvovirus B19 infections in patients with diagnosis of haematological disorders. In order to determine the diagnostic use of different markers of parvovirus B19 infection, serum specimens obtained from 79 patients with haematological disorders were tested for specific antibodies and viral DNA through the use of ELISA and PCR techniques. Evidence of parvovirus B19 infection was found in 23/79 (29.1%) patients by demonstrating viral DNA and/or specific IgM antibody. B19 infection was established in 3 of 11 patients with chronic myeloid leukemia, in 3 of 11 acute myeloid leukemia, in 2 of 11 patients with multiple myeloma, in 3 of 8 patients with Hodgkin's lymphoma, in 5 of 10 patients with non-Hodgkin's lymphoma, in 1 of 6 patients with myelodysplastic syndrome, in 4 of 11 patients with chronic lymphocytic leukemia, and in 2 of 11 patients with acute lymphocytic leukemia. In 4 of 23 positive patients, only parvovirus B19 DNA could be detected, while 7 patients were tested positive for both parvovirus B19 DNA and specific IgM. Nine patients were tested positive for both B19 DNA and specific IgG. In the remaining 3 positive patients only specific IgM could be detected. Due to the discrepancies between DNA and IgM results, the diagnostic procedures should include a search for specific DNA by PCR methods if specific IgM has been found to be negative.

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Novel PCR assay for differential detection and screening of erythrovirus B19 and erythrovirus V9

Cited by 52 publications

References 10 publications

Identification and Characterization of Persistent Human Erythrovirus Infection in Blood Donor Samples

Identification and Characterization of Persistent Human Erythrovirus Infection in Blood Donor Samples

Detection and Differentiation of Human Parvovirus Variants by Commercial Quantitative Real-Time PCR Tests

The investigation of parvovirus B19 infection in patients with haematological disorders by using PCR and ELISA techniques

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