RESUMO
B19V infection is common during childhood. It is self-limited in healthy individuals, but is often associated with transient aplastic crisis in children with sickle cell disease. The aim of this study was to estimate the prevalence and incidence of B19V infection in children with sickle cell disease screened by the Newborn Screening Program of Minas Gerais, Brazil, and followed-up at Fundação Hemominas. Serum or plasma samples from 278 patients were tested for anti-B19V IgG and IgM using commercial ELISA and for viral DNA using in-house real-time PCR assays; 127 negative-children were retested about 1 year later. The median age of children at first testing was 5.9 years (0.8-12.3). The estimated prevalence of B19V was 29.5 % (95%CI 24.1-34.9 %). The incidence of B19V in those 127 negative-children was 18.2 cases/100 patient-years. All DNA-positive samples were identified as genotype 1, except one sample, in which both genotypes 1 and 3 were identified. It was observed that the higher the child's age, the higher the probability of B19V infection. The analysis of clinical and hematological data showed a significant association of B19V infection with transient aplastic crisis and acute splenic sequestration, higher frequency of transfusions, and higher rate of hospitalization, but not with acute chest syndrome or stroke. These results emphasize the impact of B19V infection on the course of sickle cell disease. Strategies to prevent and monitor B19V infection in children with sickle cell disease should be considered to diminish its morbidity in this susceptible population.
Assuntos
Anemia Falciforme/complicações , Erythrovirus/isolamento & purificação , Infecções por Parvoviridae/epidemiologia , Adolescente , Fatores Etários , Anticorpos Antivirais/sangue , Brasil/epidemiologia , Criança , Pré-Escolar , DNA Viral/sangue , Ensaio de Imunoadsorção Enzimática , Erythrovirus/classificação , Erythrovirus/genética , Feminino , Variação Genética , Genótipo , Humanos , Imunoglobulina G/sangue , Incidência , Lactente , Masculino , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Fatores de RiscoRESUMO
Human erythrovirus B19 (EVB19) is a small, pathogenic DNA virus that has been associated with a wide range of illnesses. The primary site of replication is in bone marrow-derived erythroid progenitor cells, but EVB19 DNA has been detected in a wide range of organs. Recently, studies have linked EVB19 to thyroid cancers and other thyroid diseases. Previous studies from multiple laboratories have detected EVB19 capsid proteins in Graves' disease, Hashimoto's thyroiditis, and thyroid cancer tissues. Data on viral gene expression and mechanism of infection in the thyroid are lacking. To investigate EVB19 infection and persistence in the thyroid, previously archived adult and pediatric tissue sections were examined for EVB19 DNA, RNA, and capsid proteins, as well as EVB19 receptor P-antigen and co-receptor α5ß1 integrin. EVB19 DNA and protein were detected in a majority of tissues examined (87% and 68%, respectively). Detection was similar in adult and pediatric samples. Quantification of viral genomes revealed no significant difference in the amount of viral DNA in benign, cancerous, or metastatic thyroid tissues. EVB19 capsid RNA was detected in 67% of the tissues examined, confirming at least low-level viral gene expression. Immunohistochemical staining for P-antigen and α5ß1 detected the receptor and co-receptor most frequently on normal thyroid epithelial cells. EVB19 capsid staining could be detected in tumors lacking viral receptors. These results suggest that normal thyroid epithelial cells are the initial target for EVB19 infection in the thyroid and allow for continued persistence in both normal and cancerous thyroid tissues.
Assuntos
Adenoma/virologia , Carcinoma Papilar/virologia , Erythrovirus/genética , Infecções por Parvoviridae/virologia , Glândula Tireoide/virologia , Neoplasias da Glândula Tireoide/virologia , Adenoma/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas do Capsídeo/metabolismo , Carcinoma Papilar/metabolismo , Carcinoma Papilar/secundário , Criança , DNA Viral/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções por Parvoviridae/metabolismo , Infecções por Parvoviridae/patologia , RNA Viral/genética , Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/patologia , Adulto JovemRESUMO
Human parvovirus B19 is the only member of the genus Erythrovirus that causes human disease. Recent findings of several strains with considerable sequence divergence from B19 have suggested a new classification for parvovirus genotypes as 1 (B19), 2 (A-6 and LaLi) and 3 (V9). In their overall DNA sequence, the three genotypes differ by approximately 10%. Here, we report the isolation of a genotype-3-related strain named BR543 during a prospective study conducted in Sao Paulo, Brazil. Analysis of the nearly full-length genome sequence of BR543 indicates that this B19 variant sequence clusters with Gh2768, a strain from Ghana belonging to subtype 3b, and showed mostly synonymous substitutions.
Assuntos
Erythrovirus/genética , Sequência de Bases , Brasil , Variação Genética/genética , Genoma Viral/genética , Genótipo , Humanos , Dados de Sequência Molecular , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/genética , FilogeniaRESUMO
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/genéticaAssuntos
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
Parvovirus B19 is a single-stranded DNA virus which causes severe disease in immunocompromised patients and foetal loss in pregnant women. It is classified as an Erythrovirus and this genus also comprises two related viral genotypes (so-called LaLi/A6 (genotype 2) and V9 (genotype 3)) which appear to be immunologically indistinguishable from Parvovirus B19. Serological and nucleic acid test (NAT) systems to detect Parvovirus B19-mediated infection are commercially available; however, some NAT systems are genotype-specific. International standard preparations of Parvovirus B19 IgG and DNA have been produced for assay standardisation purposes, and to ensure consistency of assay manufacture and performance. Immunological assays, such as B-cell ELISpot, T-cell stimulation, and cytokine detection can also be used to confirm exposure to Parvovirus B19. Immunohistochemical techniques, employing commercially available monoclonal antibodies, are used to localise the virus in infected tissue and Parvovirus B19 viral antigen can also be detected in serum and plasma using antigen-specific ELISA. NAT systems have also been described to detect newly identified parvoviruses such as human bocavirus (HBoV), PARV4, and PARV5, although absolute confirmation of clinical diseases associated with these agents is required. This chapter describes the current status of detection systems for all the aforementioned parvoviruses, with particular emphasis on Erythrovirus detection by serological, NAT, and immunological approaches.
Assuntos
Erythrovirus/genética , Bocavirus Humano/genética , Parvovirinae/isolamento & purificação , Parvovirus B19 Humano/genética , Anticorpos Antivirais/isolamento & purificação , Linfócitos B/química , Primers do DNA , DNA de Cadeia Simples/genética , DNA Viral/análise , DNA Viral/genética , Ensaio de Imunoadsorção Enzimática , Erythrovirus/imunologia , Feminino , Genótipo , Humanos , Parvovirinae/genética , Parvovirus B19 Humano/imunologia , Parvovirus B19 Humano/fisiologia , Reação em Cadeia da Polimerase/métodos , Gravidez , Sensibilidade e EspecificidadeRESUMO
The transcription profile of chipmunk parvovirus (ChpPV), a tentative member of the genus Erythrovirus in the subfamily Parvovirinae of the family Parvoviridae, was characterized by transfecting a nearly full-length genome. We found that it is unique from the profiles of human parvovirus B19 and simian parvovirus, the members in the genus Erythrovirus so far characterized, in that the small RNA transcripts were not processed for encoding small non-structural proteins. However, like the large non-structural protein NS1 of the human parvovirus B19, the ChpPV NS1 is a potent inducer of apoptosis. Further phylogenetic analysis of ChpPV with other parvoviruses in the subfamily Parvovirinae indicates that ChpPV is distinct from the members in genus Erythrovirus. Thus, we conclude that ChpPV may represent a new genus in the family Parvoviridae.
Assuntos
Erythrovirus/genética , Infecções por Parvoviridae/genética , Parvoviridae/genética , Animais , Apoptose , Células COS , Chlorocebus aethiops , Erythrovirus/classificação , Humanos , Parvoviridae/classificação , Filogenia , Plasmídeos/metabolismo , Sciuridae , Transcrição GênicaAssuntos
Erythrovirus , Técnicas de Transferência de Genes , Animais , Quimera , Erythrovirus/genética , Vetores Genéticos , HumanosRESUMO
The B19 Parvovirus (B19V) has for a long time been considered as the unique human virus belonging to the genus Erythrovirus. The genetic diversity of B19V isolates has been shown to be very low. The isolation of a variant (V9 strain), with a sequence markedly distinct from that of B19V which led to attributing this classification to this family of viruses. Phylogenetic analysis of sequences of V9-related isolates indicates an organization into three well-individualized genotypes. The B19V infection can be transmitted by transfusion. In immunocompetent recipients, B19V exposure by transfusion is most often inconsequential, since a large proportion is immunized. Such an infection may have serious clinical outcome in not immunized patients with shortened red cell survival, seronegative pregnant women and immunocompromised patients. In cellular products, viral DNA detection is not performed, but a preventive strategy could be discussed for at-risk recipients. Whereas in plasma derivatives, B19V screening is performed with a threshold of 10(4)IU/ml using molecular assays. With recent data of a new classification of three genotypes within human erythrovirus, nucleic acid testing assays would be validated in accordance with the genetic variability, in order to guarantee optimal safety. Recently, a new human parvovirus (PARV4) has been discovered. The consequences on blood transfusion of this blood-borne agent and its pathogenicity are still unknown.
Assuntos
Patógenos Transmitidos pelo Sangue , Erythrovirus/genética , Infecções por Parvoviridae/transmissão , Plasma/virologia , Reação Transfusional , DNA Viral/sangue , Variação Genética , Humanos , Hospedeiro Imunocomprometido/imunologia , Infecções por Parvoviridae/prevenção & controle , Parvovirus B19 Humano/genéticaRESUMO
B19 Parvovirus (B19V) has been considered for a long period of time as the unique human virus belonging to the genus Erythrovirus. The genetic diversity of B19V isolates has been shown to be very low (<2% nucleotide divergence). The isolation of a variant (V9 strain), with a sequence markedly distinct from that of B19V (>13% nucleotide divergence) led to specify the classification of this virus family. Phylogenetic analysis of partial sequences of V9-related isolates combined with Erythrovirus sequences in sequences banks indicates an organization into three well-individualized genotypes. Analysis of the nearly full-length genome sequences show an ancient separation between the three genotypes lineages. Genotype 3 (the most ancient lineage) could have originated in Africa. The functional regions of major proteins are conserved in the three genotypes. The frequency of these genotypes is various according to studies. Genotype 1 is predominant, except in Ghana where all the described isolates were genotype 3. A prospective French study performed between 1999 and 2001 indicated that genotypes 2 and 3 viruses circulated with a significant frequency (10%). Pathogenic properties might not differ according to the genotype.
Assuntos
Erythrovirus/genética , Variação Genética , África/epidemiologia , Antígenos Virais/genética , Erythrovirus/classificação , Erythrovirus/isolamento & purificação , Europa (Continente)/epidemiologia , Genótipo , Humanos , Infecções por Parvoviridae/epidemiologia , Filogenia , Prevalência , Estados Unidos/epidemiologiaRESUMO
Parvovirus B19 (PVB19) is a member of the human erythrovirus family detected frequently in endomyocardial biopsies from patients with dilated cardiomyopathy. Human erythroviruses cluster into three genotypes 1-3 which share a high degree of homology between major structural proteins and may cause indistinguishable infections clinically and serologically. In human cardiac tissue erythrovirus genotypes other than PVB19 have not yet been reported. Three hundred seventeen consecutive patients with symptomatic dilated cardiomyopathy (median left ventricular ejection fraction: 28.6%, range 5-45%) who underwent endomyocardial biopsy for the elucidation of the etiology, were analyzed using a new consensus PCR assay designed for the detection of the three erythrovirus genotype sequences. Endomyocardial biopsies of 151 (47.6%) patients were erythrovirus-positive. Genotype 1 specific sequences were detected in 43/151 (28.5%) of positive biopsy samples, whereas genotype 2-specific sequences so far considered rare in human disease and not yet been described in human heart tissue was identified in 108/151 (71.5%) of virus-positive endomyocardial biopsies with a preference in patients above 50 years of age. In spite of younger age, systolic left ventricular dysfunction of genotype 1-positive patients was significantly reduced as compared to genotype 2-positive patients (24.4+/-10.4% vs. 31.0+/-9.5%, P=0.0001) at the initial presentation. The data show that two genetically distinct erythrovirus variants with a different age distribution are detectable in endomyocardial biopsies of patients with dilated cardiomyopathy. The erythrovirus genotype 2, not described previously in human heart tissue, is highly prevalent in the heart but the less prevalent genotype 1 is associated with more severe disturbed cardiac function.
Assuntos
Cardiomiopatia Dilatada/virologia , Erythrovirus/isolamento & purificação , Coração/virologia , Infecções por Parvoviridae/virologia , Adulto , Idoso , Sequência de Aminoácidos , Sequência de Bases , Proteínas do Capsídeo/química , Erythrovirus/genética , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Prevalência , Alinhamento de Sequência , Carga ViralRESUMO
Human erythrovirus is a minute, single-stranded DNA virus causing many diseases, including erythema infectiosum, arthropathy, and fetal death. After primary infection, the viral genomes persist in solid tissues. Besides the prototype, virus type 1, two major variants (virus types 2 and 3) have been identified recently, the clinical significance and epidemiology of which are mostly unknown. We examined 523 samples of skin, synovium, tonsil, or liver (birth year range, 1913-2000), and 1,640 sera, by qualitative and quantitative molecular assays for the DNA of human erythroviruses. Virus types 1 and 2 were found in 132 (25%) and 58 (11%) tissues, respectively. DNA of virus type 1 was found in all age groups, whereas that of type 2 was strictly confined to those subjects born before 1973 (P < 0.001). Correspondingly, the sera from the past two decades contained DNA of type 1 but not type 2 or 3. Our data suggest strongly that the newly identified human erythrovirus type 2 as well as the prototype 1 circulated in Northern and Central Europe in equal frequency, more than half a century ago, whereafter type 2 disappeared from circulation. Type 3 never attained wide occurrence in this area during the past > or =70 years. The erythrovirus DNA persistence in human tissues is lifelong and represents a source of information about our past, the Bioportfolio, which, at the individual level, provides a registry of one's infectious encounters, and at the population level, a database for epidemiological and phylogenetic analyses.
Assuntos
Vírus de DNA/genética , Erythrovirus/genética , Variação Genética/genética , Genoma Viral/genética , Expectativa de Vida , Infecções por Parvoviridae/virologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Fígado/virologia , Pessoa de Meia-Idade , Infecções por Parvoviridae/sangue , Pele/virologia , Membrana Sinovial/virologia , Fatores de TempoRESUMO
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.
Assuntos
Medula Óssea/virologia , Erythrovirus/classificação , Variação Genética , Doenças Hematológicas/virologia , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/patogenicidade , Análise de Sequência de DNA , Adolescente , Adulto , Idoso , Brasil , Criança , DNA Viral/análise , DNA Viral/isolamento & purificação , Erythrovirus/genética , Erythrovirus/isolamento & purificação , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência MolecularRESUMO
The presence of human erythrovirus DNA in 2,440 blood donations from the United Kingdom and sub-Saharan 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.
Assuntos
Doadores de Sangue , Erythrovirus/isolamento & purificação , Infecções por Parvoviridae/virologia , Sequência de Aminoácidos , Anticorpos Antivirais/sangue , Proteínas do Capsídeo/imunologia , DNA Viral/sangue , Erythrovirus/genética , Erythrovirus/imunologia , Humanos , Dados de Sequência MolecularRESUMO
Simian parvovirus (SPV) is a member of the genus Erythrovirus and is closely related to the human parvovirus B19. Natural and experimental infection of monkeys with SPV resembles B19 infection of human. We report a detailed characterization of the viral RNAs and proteins generated following transfection of cloned SPV into COS cells and SPV infection of the human erythroid progenitor line UT-7/Epo-S1. SPV and B19 are 50% identical at the nucleotide level, and although their basic transcription and protein expression profiles were generally similar, there were also significant differences. SPV pre-mRNAs contain three introns, compared to two found for B19: an additional intron was found within the capsid-coding region. RNAs in which this intron was spliced were abundant and encoded the SPV 14-kDa protein (analogous to the B19 11-kDa protein), which initiated at an AUG in the exon preceding the third intron. Unlike B19, SPV RNAs were also spliced between the donor of the first intron and the acceptor of the second intron. The third intron was additionally spliced from a portion of these molecules; these mRNAs encoded the 14-kDa protein. A portion was not spliced further and encoded VP2. Like B19, SPV has a polyadenylation signal [AAUAAA (pA)p] in the middle of the genome, which directed efficient polyadenylation of both spliced and unspliced mRNAs (encoding a putative 10-kDa protein, analogous to the B19 7.5-kDa protein, and SPV NS1, respectively). The 14-kDa protein was localized to both in the nucleus and cytoplasm.
Assuntos
Enterovirus Humano B/genética , Erythrovirus/genética , Transcrição Gênica , Animais , Northern Blotting , Células COS , Perfilação da Expressão Gênica , Inativação Gênica , Fases de Leitura Aberta , Poliadenilação , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Erythrovirus (formerly parvovirus) B19 causes a wide range of diseases in humans, including anemia due to aplastic crisis. Diagnosis of B19 infection relies on serology and the detection of viral DNA by PCR. These techniques are usually thought to detect all erythrovirus field isolates, since the B19 genome is known to undergo few genetic variations. We have detected an erythrovirus (V9) markedly different from B19 in the serum and bone marrow of a child with transient aplastic anemia. The B19 PCR assay yielded a product that hybridized only very weakly to the B19-specific probe and whose sequence diverged more from those of 24 B19 viruses (11 to 14%) than the divergence found within the B19 group (=6.65%). Restriction enzyme analysis of the V9 genome revealed that this genetic divergence extended beyond the amplified region. Interestingly, serological tests failed to demonstrate a response characteristic of acute B19 infection. V9 could be a new erythrovirus, and new diagnostic tests are needed for its detection.
Assuntos
Anemia Aplástica/virologia , Erythrovirus/genética , Erythrovirus/isolamento & purificação , Sequência de Aminoácidos , Sequência de Bases , Erythrovirus/classificação , Genoma Viral , Humanos , Dados de Sequência Molecular , FilogeniaRESUMO
Until recently, B19 virus was considered to be the only human pathogen of the genus erythrovirus. However, other non-B19 virus strains, such as V9, have now been isolated and are thought to cause infections clinically and serologically indistinguishable from those caused by B19 virus. Whereas B19 virus related isolates have a low genetic diversity of only 1-2%, nucleotide disparity of up to 12% was found for the new isolates, suggesting that non-B19 virus isolates may not be detectable using B19 virus specific PCR methods. To overcome this problem, we designed consensus primers and probes to enable the simultaneous detection of both B19 and non-B19 virus and subsequent discrimination of the two lineages by melting temperature (T(m)) analysis. A total of 196 clinical specimens, from 185 patients with a history of or an anamnesis resembling B19 virus infection, were analyzed using the consensus PCR test. Erythrovirus DNA was detected in 37 of these samples and was found to be B19 virus specific in each case, confirming previous results using B19 virus specific PCR. Although no non-B19 virus DNA was detected in any of the clinical samples tested in this study, more extensive studies are warranted. The routine use of erythrovirus consensus PCR in the diagnosis of B19 virus infection should provide valuable information on the epidemiology and clinical role of non-B19 virus isolates; its use in screening would increase the safety of blood products.
Assuntos
Erythrovirus/genética , Erythrovirus/isolamento & purificação , Parvovirus B19 Humano/genética , Parvovirus B19 Humano/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , Proteínas do Capsídeo/genética , Sequência Consenso , DNA Viral/genética , Variação Genética , Humanos , Dados de Sequência Molecular , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/virologia , Reação em Cadeia da Polimerase/estatística & dados numéricos , Sensibilidade e Especificidade , Homologia de Sequência do Ácido NucleicoRESUMO
Diagnosis of erythrovirus B19 relies on serology and the detection of viral DNA. These techniques were believed to detect all field isolates because erythrovirus B19 has been known to undergo little genetic variation (1-2%). Recently, a distinct erythrovirus isolate termed V9, markedly different from erythrovirus B19 (>11% nucleotide disparity), was isolated from a child in France suffering from transient aplastic anemia. Standard PCR assays and serological tests failed to demonstrate an acute erythrovirus B19 infection. Subsequent sequencing of the erythrovirus V9 genome shows that the nucleotide discrepancies encompass the entire genome, indicating that standard erythrovirus B19 PCR assays will not reliably detect erythrovirus V9 DNA. As a tool for studying the epidemiological role and medical importance of this erythrovirus variant, a PCR assay is described that allows simultaneous detection of, and distinction between, erythrovirus B19 and the V9 isolate. Examination of 100 erythrovirus B19 IgM positive samples as well as plasma pools representing 100,000 Danish blood donor units for the presence of B19 and V9 DNA was performed. Despite the apparent absence of erythrovirus V9 in the clinical samples at present, the DNA sequence variability demonstrates that the erythrovirus group may be more divergent than thought previously and the child harboring this isolate may herald erythrovirus V9 as a possible emerging virus.
Assuntos
Erythrovirus/isolamento & purificação , Infecções por Parvoviridae/virologia , Reação em Cadeia da Polimerase/métodos , Doadores de Sangue , Sequência Consenso , Primers do DNA/genética , DNA Viral/análise , Dinamarca/epidemiologia , Erythrovirus/genética , Humanos , Programas de Rastreamento/métodos , Infecções por Parvoviridae/epidemiologia , Valor Preditivo dos TestesRESUMO
B19 virus is a human virus belonging to the genus Erythrovirus: The genetic diversity among B19 virus isolates has been reported to be very low, with less than 2% nucleotide divergence in the whole genome sequence. We have previously reported the isolation of a human erythrovirus isolate, termed V9, whose sequence was markedly distinct (>11% nucleotide divergence) from that of B19 virus. To date, the V9 isolate remains the unique representative of a new variant in the genus Erythrovirus, and its taxonomic position is unclear. We report here the isolation of 11 V9-related viruses. A prospective study conducted in France between 1999 and 2001 indicates that V9-related viruses actually circulate at a significant frequency (11.4%) along with B19 viruses. Analysis of the nearly full-length genome sequence of one V9-related isolate (D91.1) indicates that the D91.1 sequence clusters together with but is notably distant from the V9 sequence (5.3% divergence) and is distantly related to B19 virus sequences (13.8 to 14.2% divergence). Additional phylogenetic analysis of partial sequences from the V9-related isolates combined with erythrovirus sequences available in GenBank indicates that the erythrovirus group is more diverse than thought previously and can be divided into three well-individualized genotypes, with B19 viruses corresponding to genotype 1 and V9-related viruses being distributed into genotypes 2 and 3.