Article
Evaluation of Molecular Test for the Discrimination of “Naked” DNA from Infectious Parvovirus B19 Particles in Serum and Bone Marrow Samples
Registro en:
ALVES, Arthur Daniel Rocha et al. Evaluation of Molecular Test for the Discrimination of “Naked” DNA from Infectious Parvovirus B19 Particles in Serum and Bone Marrow Samples. Viruses, v. 14, 843, p. 1 - 13, Apr. 2022.
1999-4915
10.3390/v14040843
Autor
Alves, Arthur Daniel Rocha
Langella, Barbara Barbosa
Lima, Mariana Magaldi de Souza
Coelho, Wagner Luís da Costa Nunes Pimentel
Garcia, Rita de Cássia Nasser Cubel
Cardoso, Claudete Aparecida Araújo
Marchevsky, Renato Sergio
Pinto, Marcelo Alves
Amado, Luciane Almeida
Resumen
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.