On the utility of the benzonase treatment for correct laboratory diagnosis of parvovirus B19 infection

Reber, Ulrike; Moser, Olga; Dilloo, Dagmar; Eis‐Hübinger, Anna Maria

doi:10.1016/j.jcv.2017.07.017

Cited by 6 publications

(7 citation statements)

References 5 publications

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“…A case of recurrent B19V DNAemia in a patient with hereditary spherocytosis, which was initially interpreted as viral reactivation, was clarified after confirming the absence of infectious viral particles in the patient's blood. The hypothesis was raised that the second episode of DNAemia represented the mass release of B19V DNA from the bone marrow and was, therefore, not the result of active infection [17].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Molecular Test for the Discrimination of “Naked” DNA from Infectious Parvovirus B19 Particles in Serum and Bone Marrow Samples

Alves

Langella

Lima

et al. 2022

Viruses

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Low levels of parvovirus B19 (B19V) DNA can be detected in the circulation and in different tissue of immunocompetent individuals for months or years, which has been linked to inflammatory diseases such as cardiomyopathy, rheumatoid arthritis, hepatitis, and vasculitis. However, the detection of B19V DNA does not necessarily imply that infectious virions are present. This study aimed to evaluate the method based on the Benzonase® treatment for differentiation between the infectious virions from “naked” DNA in serum and bone marrow (BM) samples to be useful for the B19V routine diagnosis. In addition, we estimated the period of viremia and DNAemia in the sera and bone marrow of nonhuman primates experimentally infected with B19V. Serum samples from ten patients and from four cynomolgus monkeys experimentally infected with B19V followed up for 60 days were used. Most of the human serum samples became negative after pretreatment; however, only decreased viral DNA loads were observed in four patients, indicating that these samples still contained the infectious virus. Reduced B19V DNA levels were observed in animals since 7th dpi. At approximately 45th dpi, B19V DNA levels were below 105 IU/mL after Benzonase® pretreatment, which was not a consequence of active B19V replication. The test based on Benzonase® pretreatment enabled the discrimination of “naked DNA” from B19V DNA encapsidated in virions. Therefore, this test can be used to clarify the role of B19V as an etiological agent associated with atypical clinical manifestations.

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Section: Introductionmentioning

confidence: 99%

Evaluation of Molecular Test for the Discrimination of “Naked” DNA from Infectious Parvovirus B19 Particles in Serum and Bone Marrow Samples

Alves

Langella

Lima

et al. 2022

Viruses

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“…16 Previously, it was demonstrated that application of endonuclease to circulating parvoviral DNA leads to its degradation, except when DNA is packaged and thus protected in viral particles during an active viral infection. 17,18 This enabled differentiation between active infection, where intact viral particles are present, and past infection, where mere parvoviral DNA remnants are released from persistence in cells. In this study, the susceptibility to degradation of B19V DNA in blood was compared between patients with confirmed DCM and a control group of well-defined recent B19V infections as a measure for viral activity.…”

Section: Introductionmentioning

confidence: 99%

“…Previously, it was demonstrated that application of endonuclease to circulating parvoviral DNA leads to its degradation, except when DNA is packaged and thus protected in viral particles during an active viral infection 17,18 . This enabled differentiation between active infection, where intact viral particles are present, and past infection, where mere parvoviral DNA remnants are released from persistence in cells.…”

Section: Introductionmentioning

confidence: 99%

Parvovirus B19 DNA detectable in hearts of patients with dilated cardiomyopathy, but absent or inactive in blood

et al. 2021

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Aims Parvovirus B19 (B19V) is often assumed to be a cause of dilated cardiomyopathy (DCM), based on the quantification of B19V DNA in endomyocardial biopsies (EMB). Whether the presence of B19V DNA correlates with active infection is still debated. Application of the enzyme endonuclease to blood samples results in degradation of B19V DNA remnants but leaves viral particles intact, which enables differentiation between active and past infection. In this study, the susceptibility to degradation by endonuclease of B19V DNA in blood was compared between DCM patients and a control group of recent B19V infections. Methods and results Twenty blood samples from 20 adult patients with DCM, who previously tested positive for B19V DNA in EMB and/or blood, were tested with B19V PCR before and after application of endonuclease to the samples. Six blood samples tested positive for B19V DNA with a mean viral load of 2.3 × 10 4 IU/mL. In five samples, B19V DNA became undetectable after endonuclease (100% load reduction); in one sample DNA load showed a 23% log load reduction (viral load before endonuclease: 9.1 × 10 4 IU/mL; after: 6.5 × 10 3 IU/mL). Presence of cardiac inflammation did not differ between patients with B19V DNAemia (1/4) and patients without B19V DNAemia (6/14) (P value = 1.0). In all 18 control samples of proven recent B19V infections, DNA remained detectable after application of endonuclease, showing only a mean log load reduction of 2.3% (mean viral load before endonuclease: 8.1 × 10 11 IU/mL; after: 8.0 × 10 11 IU/mL). Load reduction differed significantly between the DCM group and the control group; indicating the presence of intact viral particles in the control group with proven active infection and the presence of DNA remnants in the DCM group (P value = 0.000). Conclusion During recent B19V infection, viral DNA levels in blood were unaffected by endonuclease. In contrast, B19V DNA in blood in patients with DCM became undetectable or strongly reduced after application of endonuclease. Circulating viral DNA in this subset of patients with presumed parvovirus-associated DCM does not consist of intact viral particles. Viral replicative activity cannot be assumed from demonstrating B19V DNA in cardiac tissue or in blood in DCM patients.

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“…B19V virions, or only viral DNA, might also be transported between cells via extracellular vesicles, as has been shown for adenovirus ( Sims et al, 2014 ; Ipinmoroti and Matthews, 2020 ). Moreover, low-load DNaemia can be maintained for more than a year ( Musiani et al, 1995 ; Molenaar-de Backer et al, 2016 ; Reber et al, 2017 ), as naked DNA is passively released from damaged tissue or during the natural turnover of tissue cells. Reactivation of persistent B19V is not well documented but some studies have observed viremia of B19V genotype 2 (a virus that no longer circulates in the population but persists in tissues) in immunocompromised patients, suggesting that reactivation can occur ( Liefeldt et al, 2005 ; Grabarczyk et al, 2011 ; Sterpu et al, 2014 ).…”

Section: Discussionmentioning

confidence: 99%

Prevalence, Cell Tropism, and Clinical Impact of Human Parvovirus Persistence in Adenomatous, Cancerous, Inflamed, and Healthy Intestinal Mucosa

Leskinen

Gritti

et al. 2022

Front. Microbiol.

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Parvoviruses are single-stranded DNA viruses, infecting many animals from insects to humans. Human parvovirus B19 (B19V) causes erythema infectiosum, arthropathy, anemia, and fetal death, and human bocavirus (HBoV) 1 causes respiratory tract infections, while HBoV2-4 are enteric. Parvoviral genomes can persist in diverse non-permissive tissues after acute infection, but the host-cell tropism and the impact of their tissue persistence are poorly studied. We searched for parvoviral DNA in a total of 427 intestinal biopsy specimens, as paired disease-affected and healthy mucosa, obtained from 130 patients with malignancy, ulcerative colitis (UC), or adenomas, and in similar intestinal segments from 55 healthy subjects. Only three (1.6%) individuals exhibited intestinal HBoV DNA (one each of HBoV1, 2, and 3). Conversely, B19V DNA persisted frequently in the intestine, with 50, 47, 31, and 27% detection rates in the patients with malignancy, UC, or adenomas, and in the healthy subjects, respectively. Intra-individually, B19V DNA persisted significantly more often in the healthy intestinal segments than in the inflamed colons of UC patients. The highest loads of B19V DNA were seen in the ileum and colon specimens of two healthy individuals. With dual-RNAscope in situ hybridization and immunohistochemistry assays, we located the B19V persistence sites of these intestines in mucosal B cells of lymphoid follicles and vascular endothelial cells. Viral messenger RNA transcription remained, however, undetected. RNA sequencing (RNA-seq) identified 272 differentially expressed cellular genes between B19V DNA-positive and -negative healthy ileum biopsy specimens. Pathway enrichment analysis revealed that B19V persistence activated the intestinal cell viability and inhibited apoptosis. Lifelong B19V DNA persistence thus modulates host gene expression, which may lead to clinical outcomes.

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On the utility of the benzonase treatment for correct laboratory diagnosis of parvovirus B19 infection

Cited by 6 publications

References 5 publications

Evaluation of Molecular Test for the Discrimination of “Naked” DNA from Infectious Parvovirus B19 Particles in Serum and Bone Marrow Samples

Evaluation of Molecular Test for the Discrimination of “Naked” DNA from Infectious Parvovirus B19 Particles in Serum and Bone Marrow Samples

Parvovirus B19 DNA detectable in hearts of patients with dilated cardiomyopathy, but absent or inactive in blood

Prevalence, Cell Tropism, and Clinical Impact of Human Parvovirus Persistence in Adenomatous, Cancerous, Inflamed, and Healthy Intestinal Mucosa

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