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1.
Arch Virol ; 167(6): 1405-1420, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35397685

RESUMO

Coxsackievirus A19 (CV-A19) is an enterovirus belonging to the species Enterovirus C, and the prototype strain 8663 was isolated from a patient with Guillain-Barré syndrome in Japan. In this study, we determined the complete genome sequence of a CV-A19 isolate identified in a stool sample from a child with hand, foot, and mouth disease in Xinxiang, Henan, China, in 2019 and named it CV-A19 strain 2019103106/XX/CHN/2019 - 2019103106 for short. The genome of this virus consists of 7409 nucleotides, including a 6624-nucleotide open reading frame encoding a potential polyprotein precursor of 2207 amino acids. Compared with strain 8663, strain 2019103106 showed 85.1% nucleotide sequence identity in the complete genome and 85.6% identity in the VP1 coding region, reflecting their genetic divergence. Phylogenetic analysis of strain 2019103106 and other representative EV-C strains with sequences available in the GenBank database showed that CV-A19 strains could be grouped into two clusters based on the complete or 214-nucleotide partial VP1 coding regions, and 2019103106 belonged to cluster 1, with the closest relationship to CV-A19 strain SWG82 from Shandong, China. Phylogenetic trees based on the P2 and P3 coding regions highlighted the divergence between strains 2019103106 and 8663, implying that strain 2019103106 had undergone recombination. Further recombination analysis suggested that strains V18A-like CV-A1 and BBD26-like CV-A19 probably recombined to yield strain 2019103106. The present study points out the genetic diversity of CV-A19. It expands our understanding of the evolution of the CV-A19 genome, but more genome sequences of epidemic strains are needed to explain the phylogeny and evolutionary history of CV-A19 comprehensively.


Assuntos
Infecções por Coxsackievirus , Enterovirus Humano C , Doença de Mão, Pé e Boca , Criança , China/epidemiologia , Enterovirus Humano C/genética , Genoma Viral , Genômica , Doença de Mão, Pé e Boca/genética , Humanos , Nucleotídeos , Filogenia , RNA Viral/genética
2.
Virol Sin ; 37(2): 168-176, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35277374

RESUMO

Coxsackievirus A24 variant (CVA24v) is a major pathogen that causes continued outbreaks and pandemics of acute hemorrhagic conjunctivitis (AHC). In China, the first confirmed outbreak of CVA24v-related AHC occurred in Beijing in 1988, followed by another two significant outbreaks respectively in 1994 and 2007, which coincides with the three-stage dynamic distribution of AHC in the world after 1970s. To illustrate the genetic characteristics of CVA24v in different periods, a total of 23 strains were isolated from those three outbreaks and the whole genome of those isolations were sequenced and analyzed. Compared with the prototype strain, the 23 strains shared four nucleotide deletions in the 5' UTR except the 0744 strain isolated in 2007. And at the 98th site, one nucleotide insertion was found in all the strains collected from 2007. From 1994 to 2007, amino acid polarity in the VP1 region at the 25th and the 32nd site were changed. Both the 3C and VP1 phylogenetic tree indicated that isolates from 1988 and 1994 belonged to Genotype III (GIII), and 2007 strains to Genotype IV (GIV). According to the Bayesian analysis based on complete genome sequence, the most recent common ancestors for the isolates in 1988, 1994 and 2007 were respectively estimated around October 1987, February 1993 and December 2004. The evolutionary rate of the CVA24v was estimated to be 7.45 â€‹× â€‹10-3 substitutions/site/year. Our study indicated that the early epidemic of CVA24v in Chinese mainland was the GIII. Point mutations and amino acid changes in different genotypes of CVA24v may generate intensity differences of the AHC outbreak. CVA24v has been evolving constantly with a relatively rapid rate.


Assuntos
Conjuntivite Hemorrágica Aguda , Infecções por Coxsackievirus , Enterovirus Humano C , Aminoácidos/genética , Teorema de Bayes , Pequim , China/epidemiologia , Conjuntivite Hemorrágica Aguda/epidemiologia , Infecções por Coxsackievirus/epidemiologia , Surtos de Doenças , Enterovirus Humano C/genética , Humanos , Nucleotídeos , Filogenia
3.
Infect Genet Evol ; 95: 105068, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34492386

RESUMO

Coxsackievirus-A (CV-A) is a causative agent of Hand Foot Mouth Disease (HFMD) worldwide. It belongs to the Human Enterovirus genus of the family Picornaviridae. The genomics data availability of CV-A samples, isolated from human host across different continental regions, provide an excellent opportunity to study its genetic composition, diversity, and evolutionary events. The complete genome sequences of 424 CV-A isolates were analyzed through a model-based population genetic approach implemented in the STRUCTURE program. Twelve genetically distinct sub-populations were identified for CV-A isolates with a marked Fst distinction of 0.76991 (P-value = 0.00000). Besides, genetically admixed strains were characterized in the G-Id, G-IIIb clusters constituted by the CV-A12 and CV-A6 enterovirus serotypes. The serotypes depicted inter/intra-genotype recombination and episodic positive selection signatures in the structural and non-structural protein-coding regions. The observed genetic composition of CV-A samples was also deduced by the phylogenetic tree analyses, where a uniform genetic structure was inferred for most of the CV-A genotypes. However, the CV-A6 serotype samples genetically stratified into three sub-populations that may lead to the emergence of new lineages in future. These informations may implicate in planning the effective strategies to combat the coxsackievirus-A-mediated infection.


Assuntos
Enterovirus Humano C/genética , Genoma Viral , Genótipo , Evolução Molecular , Doença de Mão, Pé e Boca/virologia
4.
PLoS One ; 16(8): e0255846, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34383835

RESUMO

Human enteroviruses (EVs) comprise more than 100 types of coxsackievirus, echovirus, poliovirus and numbered enteroviruses, which are mainly transmitted by the faecal-oral route leading to diverse diseases such as aseptic meningitis, encephalitis, and acute flaccid paralysis, among others. Since enteroviruses are excreted in faeces, wastewater-based epidemiology approaches are useful to describe EV diversity in a community. In Uruguay, knowledge about enteroviruses is extremely limited. This study assessed the diversity of enteroviruses through Illumina next-generation sequencing of VP1-amplicons obtained by RT-PCR directly applied to viral concentrates of 84 wastewater samples collected in Uruguay during 2011-2012 and 2017-2018. Fifty out of the 84 samples were positive for enteroviruses. There were detected 27 different types belonging to Enterovirus A species (CVA2-A6, A10, A16, EV-A71, A90), Enterovirus B species (CVA9, B1-B5, E1, E6, E11, E14, E21, E30) and Enterovirus C species (CVA1, A13, A19, A22, A24, EV-C99). Enterovirus A71 (EV-A71) and echovirus 30 (E30) strains were studied more in depth through phylogenetic analysis, together with some strains previously detected by us in Argentina. Results unveiled that EV-A71 sub-genogroup C2 circulates in both countries at least since 2011-2012, and that the C1-like emerging variant recently entered in Argentina. We also confirmed the circulation of echovirus 30 genotypes E and F in Argentina, and reported the detection of genotype E in Uruguay. To the best of our knowledge this is the first report of the EV-A71 C1-like emerging variant in South-America, and the first report of EV-A71 and E30 in Uruguay.


Assuntos
Enterovirus Humano A/genética , Enterovirus Humano B/genética , Ligação Genética/genética , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , Enterovirus Humano A/classificação , Enterovirus Humano A/isolamento & purificação , Enterovirus Humano B/classificação , Enterovirus Humano B/isolamento & purificação , Enterovirus Humano C/classificação , Enterovirus Humano C/genética , Enterovirus Humano C/isolamento & purificação , Genótipo , Humanos , Filogenia , RNA Viral/química , RNA Viral/genética , RNA Viral/metabolismo , Estações do Ano , América do Sul , Uruguai , Águas Residuárias/virologia
5.
Int J Mol Sci ; 22(16)2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-34445134

RESUMO

Coxsackievirus A24 variant (CVA24v) is the primary causative agent of the highly contagious eye infection designated acute hemorrhagic conjunctivitis (AHC). It is solely responsible for two pandemics and several recurring outbreaks of the disease over the last decades, thus affecting millions of individuals throughout the world. To date, no antiviral agents or vaccines are available for combating this disease, and treatment is mainly supportive. CVA24v utilizes Neu5Ac-containing glycans as attachment receptors facilitating entry into host cells. We have previously reported that pentavalent Neu5Ac conjugates based on a glucose-scaffold inhibit CVA24v infection of human corneal epithelial cells. In this study, we report on the design and synthesis of scaffold-replaced pentavalent Neu5Ac conjugates and their effect on CVA24v cell transduction and the use of cryogenic electron microscopy (cryo-EM) to study the binding of these multivalent conjugates to CVA24v. The results presented here provide insights into the development of Neu5Ac-based inhibitors of CVA24v and, most significantly, the first application of cryo-EM to study the binding of a multivalent ligand to a lectin.


Assuntos
Antivirais/farmacologia , Infecções por Coxsackievirus/dietoterapia , Enterovirus Humano C/efeitos dos fármacos , Ácido N-Acetilneuramínico/farmacologia , Conjuntivite Hemorrágica Aguda/tratamento farmacológico , Conjuntivite Hemorrágica Aguda/metabolismo , Conjuntivite Hemorrágica Aguda/virologia , Infecções por Coxsackievirus/metabolismo , Infecções por Coxsackievirus/virologia , Glucose/metabolismo , Humanos , Lectinas/metabolismo , Ligantes , Polissacarídeos/metabolismo , Receptores Virais/metabolismo
6.
ACS Chem Biol ; 15(10): 2683-2691, 2020 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-32845119

RESUMO

Coxsackievirus A24 variant (CVA24v) and human adenovirus 37 (HAdV-37) are leading causative agents of the severe and highly contagious ocular infections acute hemorrhagic conjunctivitis and epidemic keratoconjunctivitis, respectively. Currently, neither vaccines nor antiviral agents are available for treating these diseases, which affect millions of individuals worldwide. CVA24v and HAdV-37 utilize sialic acid as attachment receptors facilitating entry into host cells. Previously, we and others have shown that derivatives based on sialic acid are effective in preventing HAdV-37 binding and infection of cells. Here, we designed and synthesized novel pentavalent sialic acid conjugates and studied their inhibitory effect against CVA24v and HAdV-37 binding and infection of human corneal epithelial cells. The pentavalent conjugates are the first reported inhibitors of CVA24v infection and proved efficient in blocking HAdV-37 binding. Taken together, the pentavalent conjugates presented here form a basis for the development of general inhibitors of these highly contagious ocular pathogens.


Assuntos
Adenovírus Humanos/efeitos dos fármacos , Antivirais/farmacologia , Enterovirus Humano C/efeitos dos fármacos , Ácidos Siálicos/farmacologia , Adenovírus Humanos/química , Sítios de Ligação , Enterovirus Humano C/química , Humanos , Ligação Viral/efeitos dos fármacos , Internalização do Vírus/efeitos dos fármacos
7.
Sci Rep ; 10(1): 13761, 2020 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-32792520

RESUMO

Coxsackievirus A24 variant (CVA24v) is a major causative agent of acute hemorrhagic conjunctivitis outbreaks worldwide, yet the evolutionary and transmission dynamics of the virus remain unclear. To address this, we analyzed and compared the 3C and partial VP1 gene regions of CVA24v isolates obtained from five outbreaks in Cuba between 1986 and 2009 and strains isolated worldwide. Here we show that Cuban strains were homologous to those isolated in Africa, the Americas and Asia during the same time period. Two genotypes of CVA24v (GIII and GIV) were repeatedly introduced into Cuba and they arose about two years before the epidemic was detected. The two genotypes co-evolved with a population size that is stable over time. However, nucleotide substitution rates peaked during pandemics with 4.39 × 10-3 and 5.80 × 10-3 substitutions per site per year for the 3C and VP1 region, respectively. The phylogeographic analysis identified 25 and 19 viral transmission routes based on 3C and VP1 regions, respectively. Pandemic viruses usually originated in Asia, and both China and Brazil were the major hub for the global dispersal of the virus. Together, these data provide novel insight into the epidemiological dynamics of this virus and possibly other pandemic viruses.


Assuntos
Proteínas do Capsídeo/genética , Conjuntivite Hemorrágica Aguda/epidemiologia , Infecções por Coxsackievirus/epidemiologia , Cisteína Endopeptidases/genética , Enterovirus Humano C/genética , Proteínas Virais/genética , Proteases Virais 3C , Sequência de Bases , Conjuntivite Hemorrágica Aguda/patologia , Conjuntivite Hemorrágica Aguda/transmissão , Infecções por Coxsackievirus/patologia , Infecções por Coxsackievirus/transmissão , Cuba/epidemiologia , Surtos de Doenças , Evolução Molecular , Humanos , Filogenia , Alinhamento de Sequência
8.
Arch Virol ; 165(10): 2213-2227, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32666145

RESUMO

In this study, we investigated the epidemiology and molecular characteristics of enteroviruses associated with severe hand, foot and mouth disease (HFMD) in Shenzhen, China, during 2014-2018. A total of 137 fecal specimens from patients with severe HFMD were collected. Enterovirus (EV) types were determined using real-time reverse transcription polymerase chain reaction (RT-PCR), RT nested PCR, and sequencing. Sequences were analyzed using bioinformatics programs. Of 137 specimens tested, 97 (70.8%), 12 (8.8%), and 10 (7.3%) were positive for EV-A71, coxsackievirus A6 (CVA6), and CVA16, respectively. Other pathogens detected included CVA2 (2.9%, 4/137), CVA10 (2.9%, 4/137), CVA5 (0.7%, 1/137), echovirus 6 (E6) (0.7%, 1/137) and E18 (0.7%, 1/137). The most frequent complication in patients with proven EV infections was myoclonic jerk, followed by aseptic encephalitis, tachypnea, and vomiting. The frequencies of vomiting and abnormal eye movements were higher in EV-A71-infected patients than that in CVA6-infected or CVA16-infected patients. Molecular phylogeny based on the complete VP1 gene revealed no association between the subgenotype of the virus and disease severity. Nevertheless, 12 significant mutations that were likely to be associated with virulence or the clinical phenotype were observed in the 5'UTR, 2Apro, 2C, 3A, 3Dpol and 3'UTR of CVA6. Eight significant mutations were observed in the 5'UTR, 2B, 3A, 3Dpol and 3'UTR of CVA16, and 10 significant mutations were observed in the 5'UTR, VP1, 3A and 3Cpro of CVA10. In conclusion, EV-A71 is still the main pathogen causing severe HFMD, although other EV types can also cause severe complications. Potential virulence or phenotype-associated sites were identified in the genomes of CVA6, CVA16, and CVA10.


Assuntos
Proteínas do Capsídeo/genética , Encefalite/epidemiologia , Enterovirus Humano C/genética , Doença de Mão, Pé e Boca/epidemiologia , Mioclonia/epidemiologia , Taquipneia/epidemiologia , Vômito/epidemiologia , Criança , Pré-Escolar , China/epidemiologia , Encefalite/diagnóstico , Encefalite/fisiopatologia , Encefalite/virologia , Enterovirus Humano C/classificação , Enterovirus Humano C/isolamento & purificação , Fezes/virologia , Feminino , Expressão Gênica , Genótipo , Doença de Mão, Pé e Boca/diagnóstico , Doença de Mão, Pé e Boca/fisiopatologia , Doença de Mão, Pé e Boca/virologia , Humanos , Lactente , Recém-Nascido , Masculino , Epidemiologia Molecular , Mutação , Mioclonia/diagnóstico , Mioclonia/fisiopatologia , Mioclonia/virologia , Fenótipo , Filogenia , Índice de Gravidade de Doença , Taquipneia/diagnóstico , Taquipneia/fisiopatologia , Taquipneia/virologia , Virulência , Vômito/diagnóstico , Vômito/fisiopatologia , Vômito/virologia
9.
Arch Virol ; 165(4): 1015-1018, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32052193

RESUMO

Cases of acute haemorrhagic conjunctivitis (AHC) caused by a coxsackie virus A24 variant (CV-A24v) in Mexico have been reported since 1987; however, no molecular data on the causative strains have been available. Here, we report the identification of the etiological agent responsible for the most recent AHC outbreak in southeastern Mexico (at the end of 2017) as well as the complete genome sequences of seven isolates, using next-generation sequencing (NGS). Phylogenomic analysis of the CV-A24v sequences reported here showed similarity to contemporary strains causing AHC outbreaks in French Guiana and Uganda, forming a novel clade related to genotype IV. Moreover, a specific mutational pattern in the non-structural proteins was identified in the 2017 isolates. This is the first report of genetic characterization of CV-A24v isolates obtained in Mexico.


Assuntos
Conjuntivite Hemorrágica Aguda/virologia , Infecções por Coxsackievirus/virologia , Enterovirus Humano C/isolamento & purificação , Genoma Viral , Sequência de Bases , Conjuntivite Hemorrágica Aguda/epidemiologia , Infecções por Coxsackievirus/epidemiologia , Surtos de Doenças , Enterovirus Humano C/classificação , Enterovirus Humano C/genética , Humanos , México/epidemiologia , Sequenciamento Completo do Genoma
10.
Biomed Environ Sci ; 33(11): 829-838, 2020 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-33771236

RESUMO

OBJECTIVE: To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases. METHODS: A panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A-C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID 50 per µL and copies per µL, and the newly established methods were tested in clinical specimens collected in recent years. RESULTS: The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID 50 per µL and 10 virus copies per µL, and for the complete VP1 gene was 1 CCID 50 per µL and 100 virus copies per µL, using serially-diluted virus stocks of five serotypes. As a proof-of-concept, 25 serotypes were identified and complete VP1 sequences of 23 serotypes were obtained by this system among 858 clinical specimens positive for HEVs during the past eight surveillance seasons. CONCLUSION: This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A-D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.


Assuntos
Proteínas do Capsídeo/genética , Enterovirus Humano A/genética , Enterovirus Humano B/genética , Enterovirus Humano C/genética , Enterovirus Humano D/genética , Epidemiologia Molecular/métodos , Tipagem Molecular/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Humanos
11.
Mem Inst Oswaldo Cruz ; 114: e190160, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31411312

RESUMO

Human enteroviruses (EVs) are associated with a wide spectrum of human diseases. Here we report the complete genome sequences of one EV-C99 strain and one E29 strain obtained from children suffering from acute gastroenteritis, without symptoms of enteroviral syndromes. This is the first report of EV-C99 in South America, and the second E29 genome described worldwide. Continuous surveillance on EVs is vital to provide further understanding of the circulation of new or rare EV serotypes in the country. The present study also highlights the capacity of EVs to remain in silent circulation in populations.


Assuntos
Enterovirus Humano B/genética , Enterovirus Humano C/genética , Infecções por Enterovirus/virologia , Idoso , Brasil , Pré-Escolar , Enterovirus Humano B/isolamento & purificação , Enterovirus Humano C/isolamento & purificação , Fezes/virologia , Humanos , Masculino , Filogenia , RNA Viral/genética
12.
Emerg Infect Dis ; 25(7): 1414-1416, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31211686

RESUMO

We report a case of enterovirus C105 infection in an 11-year-old girl with lower respiratory tract symptoms that was identified through the Respiratory Virus Surveillance System, which covers 30 sentinel hospitals in all 16 districts of Beijing, China. The presence of this virus strain in China confirmed its geographically wide distribution.


Assuntos
Enterovirus Humano C , Infecções por Enterovirus/diagnóstico , Infecções por Enterovirus/virologia , Biomarcadores , Criança , China/epidemiologia , Enterovirus Humano C/classificação , Enterovirus Humano C/genética , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/história , Feminino , Genes Virais , Genótipo , História do Século XXI , Humanos , Filogenia , Vigilância da População , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/história , Infecções Respiratórias/virologia , Análise de Sequência de DNA
13.
Arch Virol ; 164(8): 2183-2186, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31119477

RESUMO

Enterovirus C96 (EV-C96) is a newer member of the species Enterovirus C. In this study, we determined the complete genome sequences of three EV-C96 isolates, one recovered from domestic sewage in 2013 and the other two isolated during surveillance of acute flaccid paralysis cases in 1991 and 2009, respectively. The complete genome sequences of these isolates were 75.6-84.2% identical to each other, 75.1-81.8% identical to the prototype strain, and 75.0-91.5% identical to other previously reported strains. Phylogenetic analysis of VP1 sequences revealed a high degree of genetic divergence among currently available EV-C96 sequences in the GenBank database, with an overall mean p-distance of 0.176. It is interesting to note that the 1991 strain 127/SD/CHN/1991 is the earliest EV-C96 isolate so far. Although EV-C96 is not frequently isolated during enterovirus surveillance, its great genetic diversity and the above findings suggest that this serotype has been circulating in China for many years.


Assuntos
Enterovirus Humano C/genética , Enterovirus Humano C/isolamento & purificação , Infecções por Enterovirus/virologia , Genoma Viral/genética , China , Humanos , Filogenia , RNA Viral/genética , Análise de Sequência de DNA/métodos , Sequenciamento Completo do Genoma/métodos
14.
PLoS Negl Trop Dis ; 13(4): e0007335, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31002713

RESUMO

Enteroviruses (EVs) are among the most common viruses infecting humans worldwide but only a few Non-Polio Enterovirus (NPEV) isolates have been characterized in the Democratic Republic of Congo (DR Congo). Moreover, circulating vaccine-derived polioviruses (PVs) [cVDPVs] isolated during multiple outbreaks in DR Congo from 2004 to 2018 have been characterized so far only by the sequences of their VP1 capsid coding gene. This study was carried to i) investigate the circulation and genetic diversity of NPEV and polio vaccine isolates recovered from healthy children and Acute Flaccid Paralysis (AFP) patients, ii) evaluate the occurrence of genetic recombination among EVs belonging to the Enterovirus C species (including PVs) and iii) identify the virological factors favoring multiple emergences of cVDPVs in DR Congo. The biological material considered in this study included i) a collection of 91 Sabin-like PVs, 54 cVDPVs and 150 NPEVs isolated from AFP patients between 2008 and 2012 in DR Congo and iii) a collection of 330 stool specimens collected from healthy children in 2013 in the Kasai Oriental and Maniema provinces of DR Congo. Studied virus isolates were sequenced in four distinct sub-genomic regions 5'-UTR, VP1, 2CATPase and 3Dpol. Resulting sequences were compared through comparative phylogenetic analyses. Virus isolation showed that 19.1% (63/330) healthy children were infected by EVs including 17.9% (59/330) of NPEVs and 1.2% (4/330) of type 3 Sabin-like PVs. Only one EV-C type, EV-C99 was identified among the NPEV collection from AFP patients whereas 27.5% of the 69 NPEV isolates typed in healthy children belonged to the EV-C species: CV-A13 (13/69), A20 (5/69) and A17 (1/69). Interestingly, 50 of the 54 cVDPVs featured recombinant genomes containing exogenous sequences in at least one of the targeted non-structural regions of their genomes: 5'UTR, 2CATPase and 3Dpol. Some of these non-vaccine sequences of the recombinant cVDPVs were strikingly related to homologous sequences from co-circulating CV-A17 and A20 in the 2CATPase region as well as to those from co-circulating CV-A13, A17 and A20 in the 3Dpol region. This study provided the first evidence uncovering CV-A20 strains as major recombination partners of PVs. High quality AFP surveillance, sensitive environmental surveillance and efficient vaccination activities remain essential to ensure timely detection and efficient response to recombinant cVDPVs outbreaks in DR Congo. Such needs are valid for any epidemiological setting where high frequency and genetic diversity of Coxsackieviruses A13, A17 and A20 provide a conducive viral ecosystem for the emergence of virulent recombinant cVDPVs.


Assuntos
Enterovirus Humano C/genética , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Evolução Molecular , Poliovirus/genética , Recombinação Genética , Linhagem Celular , Criança , República Democrática do Congo/epidemiologia , Humanos , Filogenia , Vacina Antipólio Oral , Estudos Prospectivos , RNA Viral/genética , RNA Viral/isolamento & purificação , Estudos Retrospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência
15.
Arch Virol ; 164(5): 1489-1492, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30877454

RESUMO

In this study, human enterovirus C117 (EV-C117) was detected in a 3-month-old boy diagnosed with pneumonia in China. A phylogenetic analysis showed that this strain was genetically closer to the Lithuanian strain than to the USA strain.


Assuntos
Enterovirus Humano C/genética , Infecções por Enterovirus/diagnóstico , Genoma Viral/genética , Pneumonia Viral/virologia , Sequência de Bases , China , Enterovirus Humano C/classificação , Enterovirus Humano C/isolamento & purificação , Infecções por Enterovirus/virologia , Humanos , Lactente , Masculino , Filogenia , Pneumonia Viral/diagnóstico , RNA Viral/genética , Análise de Sequência de RNA
16.
Arch Virol ; 164(4): 1181-1185, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30725183

RESUMO

A large outbreak (over 200,000 cases) of acute hemorrhagic conjunctivitis (AHC) took place in Brazil during the summer of 2017/2018, seven years after a nationwide epidemic, which occurred in 2011. To identify the etiological agent, 80 conjunctival swabs from patients with a clinical presentation suggestive of AHC were analyzed at the national enterovirus laboratory. Real-time RT-PCR for human enteroviruses was performed, and enterovirus RNA was detected in 91.25% (73/80) of the specimens. Twenty-nine swab fluids were used to inoculate cell cultures (RD and Hep2C), and 72.4% (21/29) yielded a cytopathic effect. Genotype IV coxsackievirus A24v (CV-A24v) was identified as the causative agent of the outbreak. Phylogenetic analysis based on the VP1 gene revealed that Brazilian isolates were genetically related to strains that caused an outbreak in French Guiana in 2017. Our results show the re-emergence of CV-A24v causing AHC outbreaks in Brazil between the end of 2017 and the beginning of 2018.


Assuntos
Conjuntivite Hemorrágica Aguda/virologia , Infecções por Coxsackievirus/virologia , Enterovirus Humano C/isolamento & purificação , Adulto , Brasil/epidemiologia , Conjuntivite Hemorrágica Aguda/epidemiologia , Infecções por Coxsackievirus/epidemiologia , Surtos de Doenças , Enterovirus Humano C/classificação , Enterovirus Humano C/genética , Enterovirus Humano C/fisiologia , Feminino , Genótipo , Humanos , Masculino , Filogenia , Adulto Jovem
17.
J Virol Methods ; 264: 38-43, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30447245

RESUMO

Polioviruses are members of the Enterovirus C species and asymptomatic fecal shedding allows for their transmission and persistence in a community, as well as the emergence of vaccine-derived polioviruses. Using three serotype-specific real-time RT-PCR (rRT-PCR) assays, the shedding and circulation of oral poliovirus vaccine (OPV) strains was previously investigated in a prospective cohort of Mexican children, their contacts, and nearby sewage. Subsequently, a deep sequencing approach targeting the P1 genomic region was applied to characterize OPV strains previously detected by rRT-PCR. Amplifiable RNA was obtained for sequencing from 40.3% (58/144) of stool samples and 71.4% (15/21) of sewage using nucleic acids extracted directly from primary rRT-PCR-positive specimens. Sequencing detected one or more OPV serotypes in 62.1% (36/58) of stool and 53.3% (8/15) of sewage samples. All stool and sewage samples in which poliovirus was not detected by deep sequencing contained at least one non-polio enterovirus C (NPEV-C) strain. To improve screening specificity, a modified, two-step, OPV serotype-specific multiplex rRT-PCR was evaluated. In stool specimens, the overall agreement between the original assays and the multiplex was 70.3%. By serotype, the overall agreement was 95.7% for OPV serotype-1 (S1), 65.6% for S2, and 96.1% for S3. Furthermore, most original rRT-PCR positive/multiplex rRT-PCR negative results were collected in the summer and fall months, consistent with NPEV-C circulation patterns. In conclusion, this deep sequencing approach allowed for the characterization of OPV sequences directly from clinical samples and facilitated the implementation of a more specific multiplex rRT-PCR for OPV detection and serotyping.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Poliovirus/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Enterovirus Humano C/genética , Enterovirus Humano C/isolamento & purificação , Fezes/virologia , Humanos , Poliovirus/genética , Estudos Prospectivos , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Sorogrupo , Esgotos/virologia
18.
Mem. Inst. Oswaldo Cruz ; 114: e190160, 2019. graf
Artigo em Inglês | LILACS | ID: biblio-1040614

RESUMO

Human enteroviruses (EVs) are associated with a wide spectrum of human diseases. Here we report the complete genome sequences of one EV-C99 strain and one E29 strain obtained from children suffering from acute gastroenteritis, without symptoms of enteroviral syndromes. This is the first report of EV-C99 in South America, and the second E29 genome described worldwide. Continuous surveillance on EVs is vital to provide further understanding of the circulation of new or rare EV serotypes in the country. The present study also highlights the capacity of EVs to remain in silent circulation in populations.


Assuntos
Humanos , Masculino , Pré-Escolar , Idoso , RNA Viral/genética , Enterovirus Humano B/genética , Enterovirus Humano C/genética , Infecções por Enterovirus/virologia , Filogenia , Brasil , Enterovirus Humano B/isolamento & purificação , Enterovirus Humano C/isolamento & purificação , Fezes/virologia
19.
Sci Rep ; 8(1): 13357, 2018 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-30190558

RESUMO

Coxsackievirus A10 (CV-A10) associated with Hand, foot, and mouth disease (HFMD) cases emerged increasingly in recent years. In this study, the samples from nation-wide HFMD surveillance, including 27 out of 31 provinces in China were investigated, and the continuous and extensive virological surveillance, covered 13 years, were conducted to provide a comprehensive molecular characterization analysis of CV-A10. 855 CV-A10 viruses (33 severe cases included), were isolated from HFMD children patients during 2009 to 2016 in China. 164 representative sequences from these viruses, together with 117 CV-A10 sequences downloaded from GenBank based on entire VP1 were recruited in this study. Two new genotypes (F and G) and two sub-genotypes (C1 and C2) were identified. Among 264 Chinese sequences, 9 of them were genotype B, 8 of them were C1, and the other (247) were C2, the predominant sub-genotype in China since 2012. Chinese C2 viruses showed obvious temporal characteristics and can be divided into 3 clusters (cluster 1~3). Cluster 3 viruses was circulating extensively during 2014 and 2016 with more severe cases. It is very necessary and important to continuously conduct the extensive virological surveillance for CV-A10, and further evolutionary studies will provide more evidence on its evolution and virulence.


Assuntos
Enterovirus Humano C/genética , Evolução Molecular , Genótipo , Doença de Mão, Pé e Boca/epidemiologia , Doença de Mão, Pé e Boca/genética , Adolescente , Criança , Pré-Escolar , China , Feminino , Humanos , Masculino
20.
J Neuropathol Exp Neurol ; 77(8): 661-664, 2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-29850899

RESUMO

Thymomas are associated with autoantibody formation. The most common are anti-acetylcholine receptor antibodies, which correspond to myasthenia gravis (MG). Other autoantibodies, such as antistriational antibodies, can occur, but their relation to clinical syndromes is frequently uncertain. The etiology of antistriational antibodies is also poorly understood. In this case, a 61-year-old man with a history of thymoma was admitted with respiratory failure. The patient was positive for anti-acetylcholine receptor antibodies and antistriational antibodies. He developed cardiogenic shock and died within 2 days despite aggressive therapy. Laboratory studies revealed elevated cardiac enzymes and marked IgG elevation against Coxsackie A virus serotypes 9 and 24. Subclinical IgG elevations against additional Coxsackie A and Coxsackie B virus serotypes were also noted. Autopsy revealed lymphohistiocytic infiltrates with multinucleated giant cells in the myocardium and skeletal muscles, including the diaphragm. Giant cell polymyositis and myocarditis is a rare, lethal complication in patients with thymoma and MG. The pathogenesis is uncertain. An autoimmune process, possibly elicited by antistriational antibodies, has been suggested. The coexistence of antistriational antibodies and Coxsackie viral serologies has not been reported. This case may suggest that giant cell polymyositis and myocarditis in patients with thymoma and MG is a postviral autoimmune process.


Assuntos
Enterovirus/metabolismo , Miastenia Gravis/sangue , Miocardite/sangue , Polimiosite/sangue , Timoma/sangue , Neoplasias do Timo/sangue , Enterovirus/isolamento & purificação , Enterovirus Humano B/isolamento & purificação , Enterovirus Humano B/metabolismo , Enterovirus Humano C/isolamento & purificação , Enterovirus Humano C/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Miastenia Gravis/complicações , Miastenia Gravis/diagnóstico , Miocardite/complicações , Miocardite/diagnóstico , Polimiosite/complicações , Polimiosite/diagnóstico , Timoma/complicações , Timoma/diagnóstico , Neoplasias do Timo/complicações , Neoplasias do Timo/diagnóstico , Carga Viral/fisiologia
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