Assuntos
Antígenos CD36/sangue , Erythrovirus/classificação , Erythrovirus/fisiologia , Células-Tronco Hematopoéticas/citologia , Replicação Viral , Adulto , Antígenos CD34/sangue , Antígenos CD36/imunologia , Proteínas do Capsídeo/metabolismo , Hipóxia Celular/fisiologia , Erythrovirus/genética , Erythrovirus/imunologia , Erythrovirus/metabolismo , Feminino , Sangue Fetal/citologia , Células-Tronco Hematopoéticas/virologia , Humanos , Fator 1 Induzível por Hipóxia/metabolismo , Técnicas In Vitro , Oxigênio , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/genética , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Gravidez , Regiões Promotoras Genéticas/genética , Regiões Promotoras Genéticas/imunologia , Fatores de Tempo , Proteínas não Estruturais Virais/metabolismo , Vírion/metabolismoRESUMO
Variant samples from the three genotypes of erythroviruses have already been detected using sequencing as methodology for analysis. This study aimed to investigate the efficacy of single-stranded conformation polymorphism (SSCP) analysis and heteroduplex mobility assay (HMA) as methodologies to detect human erythrovirus variants, using their VP1 unique region sequences. Clinical samples and plasmids of PVBAUA, A6, LaLi, V9Gh3051, and D91.1 erythrovirus variants as prototypes of the three genotypes were used. SSCP analysis was able to distinguish all divergences among the plasmids, including the two mutation points between LaLi and A6 plasmids that led to distinct electrophoresis mobility patterns. Although HMA analysis was unabled to detect two mutation points between LaLi and A6, it enabled the differentiation among all other plasmids that revealed specific electrophoresis patterns, with high-enough sensibility to detect 1.5% nucleotide substitutions. When 57 clinical samples were analyzed, 33 of them presented an identical pattern to PVBAUA by HMA and SSCP analyses, two of them were sequenced and presented an identical sequence in relation to PVBAUA. Another pattern was found for 21 samples. Among these, two samples were sequenced, revealing one mutation point in relation to PVBAUA, while each one of the three remaining samples presented a distinct pattern, showing two or three mutations in relation to PVBAUA by sequencing. HMA and SSCP analyses were suggested as methodologies suited for detecting genetic mutations of human erythroviruses in developing countries because of their practicability and minor costs for reagents and equipment.
Assuntos
DNA Viral/genética , Erythrovirus/classificação , Erythrovirus/genética , Análise Heteroduplex/métodos , Infecções por Parvoviridae/virologia , Polimorfismo Conformacional de Fita Simples , Adolescente , Adulto , Sequência de Bases , Criança , Pré-Escolar , Erythrovirus/isolamento & purificação , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , Mutação Puntual , Sensibilidade e Especificidade , Análise de Sequência de DNA , Proteínas Estruturais Virais/genéticaRESUMO
The presence of erythrovirus infections was investigated by PCR with bone marrow samples of patients with various parvovirus B19-related hematological symptoms. Erythrovirus DNA was found in 17.3% (12/69) of patients. Phylogenetic analysis revealed that five strains cluster with genotype 1, one clusters with genotype 2, and six cluster with genotype 3. Our study is the first to document the presence of the three erythrovirus genotypes in Brazil.