RESUMO
Norovirus is a major cause of acute diarrheal disease (ADD) outbreaks worldwide. In the present study, we investigated an ADD outbreak caused by norovirus in several municipalities of Santa Catarina state during the summer season, southern Brazil in 2023. As of the 10th epidemiological week of 2023, approximately 87 000 ADD cases were reported, with the capital, Florianópolis, recording the highest number of cases throughout the weeks. By using RT-qPCR and sequencing, we detected 10 different genotypes, from both genogroups (G) I and II. Some rare genotypes were also identified. Additionally, rotavirus and human adenovirus were sporadically detected among the ADD cases. Several features of the outbreak suggest that sewage-contaminated water could played a role in the surge of ADD cases. Storm events in Santa Catarina state that preceded the outbreak likely increased the discharge of contaminated wastewater and stormwater into water bodies, such as rivers and beaches during a high touristic season in the state. Climate change-induced extreme weather events, including intensified rainfall and frequent floods, can disturb healthcare and sanitation systems. Implementing public policies for effective sanitation, particularly during peak times, is crucial to maintain environmental equilibrium and counter marine pollution.
Assuntos
Infecções por Caliciviridae , Gastroenterite , Norovirus , Humanos , Norovirus/genética , Brasil/epidemiologia , Surtos de Doenças , Genótipo , Água , Infecções por Caliciviridae/epidemiologia , FezesRESUMO
We report on the occurrence and diversity of noroviruses in children (younger than 5 years old of age) from a low-income urban area in Rio de Janeiro, Brazil. Sixty-one stool specimens collected from children between 1 and 4 years old with acute diarrhoeic episodes (ADE) and non-ADE were investigated. RT-qPCR and sequencing of PCR products after conventional RT-PCR analysis were performed. Noroviruses were detected in 29 (47.5%) samples: 21 (46.7%) from cases with ADE and 8 (50%) from non-ADE cases. Molecular characterization showed 10 different genotypes circulating in this community between November 2014 and April 2018.
Assuntos
Gastroenterite/virologia , Variação Genética/genética , Norovirus/genética , Brasil , Pré-Escolar , Fezes/virologia , Gastroenterite/diagnóstico , Genótipo , Humanos , Lactente , Norovirus/isolamento & purificação , Filogenia , Pobreza , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNARESUMO
Group A rotavirus (RVA) genotype G12 is habitually associated with diarrhea disease (DD) in African children and recently its detection has increased worldwide. A total of 970 stool samples collected from individuals with DD in the Northeastern, Southeastern, and Southern Brazilian regions, Eastern coast, were analyzed and 321 (33%) were positive for RVA and of these, 241 (75%) genotyped as G12P[8]. The rate of RVA positivity was higher among children aged 5-10 years old (60%). All RVA infections observed in adults aged >21 years were G12P[8] (n = 27) showing that this genotype affected older age groups during the year of 2014 in Brazil. Phylogenetic analysis of VP7 and VP8* G12P[8] strains demonstrated an elevated similarity among Brazilian and G12-III prototypes strains circulating worldwide recently, suggesting that this lineage is associated with the global spread of the G12 genotype, considered as the 6th most prevalent human RVA genotype nowadays; while other G12 lineages remain sporadically detected and usually detected in association with other P genotypes. VP8* analysis revealed that Brazilian strains belong to P[8]-3 lineage, the single P[8] lineage presently detected in the country. No major nucleotide/amino acid disparities were observed among strains recovered from children and adults for VP7 and VP8* genes. These data are essential to support the surveillance studies, particularly in countries where the RVA vaccine was introduced in their National Immunization Program enabling identification of potential alterations in the epidemiological profile that can impact its efficacy in vaccination programs. J. Med. Virol. 89:64-70, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Antígenos Virais/genética , Proteínas do Capsídeo/genética , Genótipo , Proteínas de Ligação a RNA/genética , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Rotavirus/classificação , Rotavirus/genética , Proteínas não Estruturais Virais/genética , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , Criança , Pré-Escolar , Fezes/virologia , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Filogenia , Rotavirus/isolamento & purificação , Adulto JovemRESUMO
This study aimed to assess two homogenization methods to recover norovirus from Minas artisanal cheese (MAC) made with raw bovine milk obtained from four microregions of the Minas Gerais state, Brazil, with different ripening times and geographical and abiotic characteristics. For this purpose, 33 fiscal samples were artificially contaminated with norovirus GI and GII, and Mengovirus (MgV), used as an internal process control (IPC). TRIzol® reagent and Proteinase K homogenization methods were evaluated for all samples were then subjected to RNA extraction using viral magnetic beads and RT-qPCR Taqman® for viral detection/quantification. Proteinase K method showed better efficiency results for both norovirus GI and GII, with means recovery efficiency of 45.7% (95% CI 34.3-57.2%) and 41.4% (95% CI 29.1-53.6%), respectively, when compared to TRIzol method (16.6% GI, 95% CI 8.4-24.9%, and 12.3% GII, 95% CI 7.0-17.6%). The limits of detection for norovirus GI and GII for this method were 101GC/g and 103GC/g, respectively, independent of cheese origin. MgV was detected and revealed in 100% success rate in all types of cheese, with mean recovery efficiency of 25.6% for Proteinase K, and 3.8% for the TRIzol method. According to cheese origin, Triangulo Mineiro MAC had the highest mean recovery rates for the three viral targets surveyed (89% GI, 87% GII, and 51% MgV), while Serro MAC showed the lowest rates (p < 0.001). Those results indicate that the proteinase K adapted method is suitable for norovirus GI and GII detection in MAC and corroborated MgV as an applicable IPC to be used during the process.
Assuntos
Queijo , Contaminação de Alimentos , Leite , Norovirus , Queijo/virologia , Norovirus/isolamento & purificação , Norovirus/genética , Norovirus/classificação , Animais , Leite/virologia , Bovinos , Brasil , Contaminação de Alimentos/análise , RNA Viral/isolamento & purificação , RNA Viral/genética , RNA Viral/análise , Fast Foods/virologia , Fast Foods/análiseRESUMO
Rotavirus A (RVA) remains a leading cause of acute gastroenteritis (AGE) hospitalizations in children worldwide. During the COVID-19 pandemic, a reduction in vaccination coverage in Brazil and elsewhere was observed, and some reports have demonstrated a reduction in AGE notifications during the pandemic. This study aims to investigate the diversity and prevalence of RVA genotypes in children and adults presenting with AGE symptoms in Brazil during the COVID-19 pandemic between 2020 and 2022. RVA was screened using RT-qPCR; then, G and P genotypes were characterized using one-step multiplex RT-PCR. A total of 2173 samples were investigated over the three-year period, and we detected RVA in 7.7% of samples (n = 167), being 15.5% in 2020, 0.5% in 2021, and 13.8% in 2022. Higher RVA prevalence was observed in the Northeastern region (19.3%) compared to the Southeastern (6.1%) and Southern regions (5.5%). The most affected age group was children aged between 0 and 6 months old; however, this was not statistically significant. Genotyping and phylogenetic analysis identified the emergence of G6P[8] during the period; moreover, it was detected in 10.6% of samples in 2020 and in 83.5% in 2022. In contrast, the prevalence of G3P[8], the previous dominant genotype, decreased from 72.3% in 2020 to 11.3% in 2022. We also identified unusual strains, such as G3P[9] and G9P[4], being sporadically detected during the period. This is the first report on the molecular epidemiology and surveillance of RVA during the COVID-19 pandemic period in Brazil. Our study provides evidence for the importance of maintaining high and sustainable levels of vaccine coverage to protect against RVA disease. Furthermore, it highlights the need to maintain nationwide surveillance in order to monitor future trends and changes in the epidemiology of RVA in Brazil.
Assuntos
COVID-19 , Rotavirus , Adulto , Criança , Humanos , Recém-Nascido , Lactente , Rotavirus/genética , Brasil/epidemiologia , COVID-19/epidemiologia , Pandemias , Filogenia , GenótipoRESUMO
Norovirus stands out as a leading cause of acute gastroenteritis (AGE) worldwide, affecting all age groups. In the present study, we investigated fecal samples from medically attended AGE patients received from nine Brazilian states, from 2019 to 2022, including the COVID-19 pandemic period. Norovirus GI and GII were detected and quantified using RT-qPCR, and norovirus-positive samples underwent genotyping through sequencing the ORF1/2 junction region. During the four-year period, norovirus prevalence was 37.2%, varying from 20.1% in 2020 to 55.4% in 2021. GII genotypes dominated, being detected in 92.9% of samples. GII-infected patients had significantly higher viral concentrations compared to GI-infected patients (median of 3.8 × 107 GC/g and 6.7 × 105 GC/g, respectively); and patients aged >12-24 months showed a higher median viral load (8 × 107 GC/g) compared to other age groups. Norovirus sequencing revealed 20 genotypes by phylogenetic analysis of RdRp and VP1 partial regions. GII.4 Sydney[P16] was the dominant genotype (57.3%), especially in 2019 and 2021, followed by GII.2[P16] (14.8%) and GII.6[P7] (6.3%). The intergenogroup recombinant genotype, GIX.1[GII.P15], was detected in five samples. Our study is the first to explore norovirus epidemiology and genotype distribution in Brazil during COVID-19, and contributes to understanding the epidemiological dynamics of norovirus and highlighting the importance of continuing to follow norovirus surveillance programs in Brazil.
RESUMO
The introduction of rotavirus A (RVA) vaccines has considerably reduced the RVA-associated mortality among children under 5 years of age worldwide. The ability of RVA to reassort gives rise to different combinations of surface proteins G (glycoprotein, VP7) and P (protease sensitive, VP4) RVA types infecting children. During the epidemiological surveillance of RVA in the Northwest Amazon region, an unusual rotavirus genotype G6P[8] was detected in feces of a 2-year-old child with acute gastroenteritis (AGE) that had been vaccinated with one dose of Rotarix® (RV1). The G6P[8] sample had a DS-1-like constellation with a Wa-like VP3 gene mono-reassortment similar to equine-like G3P[8] that has been frequently detected in Brazil previously. The results presented here reinforce the evolutionary dynamics of RVA and the importance of constant molecular surveillance.
RESUMO
Human adenovirus (HAdV) types F40/41 have long been recognized as major viral agents of acute gastroenteritis (AGE) in children. Despite this, studies on HAdV molecular epidemiology are sparse, and their real impact is likely under-estimated. Thus, our goal was to investigate HAdV incidence, enteric and non-enteric types circulation, co-detections with rotavirus and norovirus and DNA shedding in stool samples from inpatients and outpatients from eleven Brazilian states. During the three-year study, 1012 AGE stool samples were analysed by TaqMan-based qPCR, to detect and quantify HAdV. Positive samples were genotyped by partial sequencing of the hexon gene followed by phylogenetic analysis. Co-detections were accessed by screening for rotavirus and norovirus. Overall, we detected HAdV in 24.5% of single-detected samples (n = 248), with a prevalence of type F41 (35.8%). We observed a higher incidence in children between 6 to 24 months, without marked seasonality. Additionally, we observed a statistically higher median viral load among single-detections between enteric and non-enteric types and a significantly lower HAdV viral load compared to rotavirus and norovirus in co-detections (p < 0.0001). Our study contributes to the knowledge of HAdV epidemiology and reinforces the need for the inclusion of enteric types F40/41 in molecular surveillance programs.
Assuntos
Adenovírus Humanos , Gastroenterite , Norovirus , Rotavirus , Adenovírus Humanos/genética , Brasil/epidemiologia , Criança , Fezes , Gastroenterite/epidemiologia , Humanos , Norovirus/genética , Filogenia , Rotavirus/genéticaRESUMO
OBJECTIVE: The aim of the present study was to characterize 24 isolates of group C rotavirus (RV-C) by analysis of DNA sequences of the VP4, VP6, VP7 and NSP4 genes of several Brazilian states. METHODS: All RV-C strains were confirmed by polyacrylamide gel electrophoresis and were characterized by sequence and phylogenetic analysis of the genes encoding NSP4, VP6, VP4, and VP7 proteins. RESULTS: Analysis of the NSP4 gene from Brazilian strains revealed that the isolates are more closely related to each other than to those of other strains previously published. The VP6 gene showed high homology to Indian strains. The sequences of VP4 and VP7 genes showed two lineages circulating in Brazil in the same region and at the same time. CONCLUSION: These results confirm the transmission of RV-C in Brazil. RV-C infection appears to occur occasionally despite the low detection rate of the virus.
Assuntos
RNA Viral/genética , Infecções por Rotavirus/virologia , Rotavirus/classificação , Rotavirus/isolamento & purificação , Adolescente , Adulto , Brasil , Criança , Pré-Escolar , Análise por Conglomerados , Feminino , Genótipo , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , Rotavirus/genética , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Proteínas Virais/genética , Adulto JovemRESUMO
Worldwide, rotavirus (RVA) and norovirus are considered major etiological agents of acute gastroenteritis (AGE) in pediatric population admitted to hospitals. This study describes the investigation of nosocomial infections caused by emergent RVA and norovirus strains reported at a pediatric hospital in southern Brazil in May 2019. This outbreak affected 30 people among children and adults. Nine stool samples (eight children and one nurse) were obtained and analyzed by RT-qPCR to detect and quantify RVA and norovirus. Positive samples were genotyped by sequencing and subjected to phylogenetic analysis. We detected RVA in 44.4% (4/9) and norovirus in 55.5% (5/9) at high viral loads, ranging from 3.5 × 107 to 6.1 × 107 and 3.2 × 102 to 3.2 × 109 genome copies/g of stool, respectively. Co-infections were not observed. RVA VP4 and VP7 gene sequencing in combination with polyacrylamide gel electrophoresis identified the circulation of equine-like G3P[8] DS-1-like, and the partial sequencing of the other nine genes revealed that strains possessed I2-R2-C2-M2-A2-N1-T2-E2-H2 genotype background. The emergent recombinant norovirus variant, GII.4 Sydney[P16], was identified by ORF1-2 sequencing. Active surveillance and effective prevention measures should be constantly reinforced to avoid the spread of nosocomial viral infections into hospitals, which could severely affect pediatric patients admitted with underlying health conditions.
Assuntos
Infecções por Caliciviridae , Infecção Hospitalar , Gastroenterite , Norovirus , Infecções por Rotavirus , Rotavirus , Animais , Brasil/epidemiologia , Infecções por Caliciviridae/epidemiologia , Criança , Infecção Hospitalar/epidemiologia , Surtos de Doenças , Fezes , Gastroenterite/epidemiologia , Genoma Viral , Genótipo , Cavalos , Hospitais Pediátricos , Humanos , Norovirus/genética , Filogenia , Rotavirus/genética , Infecções por Rotavirus/epidemiologiaRESUMO
OBJECTIVES: To verify the frequency of viruses causing acute gastroenteritis (AGE) in association with the histo-blood group antigen (HBGA) and Rotarix™ vaccination coverage in children from the Amazon region. DESIGN: Fecal and saliva samples were collected from children with AGE (n = 485) and acute respiratory infection (ARI) (n = 249) clinical symptoms. Rotavirus A (RVA), norovirus, human adenovirus (HAdV), and sapovirus (SaV) were verified in feces by molecular detection. Saliva samples were used for HBGA phenotyping/FUT3 genotyping. Blood group types, clinical aspects and Rotarix™ RVA vaccination data were recorded. RESULTS: Norovirus remained the most prevalently detected cause of AGE (38%, 184/485 and ARI 21.3%, 53/249). High HAdV frequencies were observed in AGE children (28.6%, 139/485) and ARI children (37.3%, 93/249). RVA was the third most prevalent virus causing AGE (22.7%, 110/485 and ARI 19.3%, 48/249) and a low RV1 coverage (61%, 448/734) was verified. The SaV frequencies were lower (7.2%, 35/485 for AGE and 6.8%, 17/249 for ARI). Secretor children were HBGA susceptible to HAdV infection (OR 1.5, 95% CI 1.0-2.3; P = 0.04) but not to RVA, norovirus or SaV infection. CONCLUSIONS: Norovirus could be considered the main etiological agent of AGE. No association was verified for HBGA susceptibility to RVA, norovirus and SaV. Secretor children showed a slight susceptibility to HAdV infection and the Le (a-b-) heterogeneous SNPs on the FUT3 gene.
Assuntos
Gastroenterite/virologia , Viroses/epidemiologia , Doença Aguda , Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/isolamento & purificação , Antígenos de Grupos Sanguíneos/análise , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Pré-Escolar , Fezes/virologia , Feminino , Fucosiltransferases/genética , Gastroenterite/epidemiologia , Gastroenterite/genética , Genótipo , Humanos , Lactente , Masculino , Norovirus/isolamento & purificação , Polimorfismo de Nucleotídeo Único , Infecções Respiratórias/virologia , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Vacinas contra Rotavirus , Saliva , Sapovirus/isolamento & purificação , América do Sul/epidemiologia , Vacinas AtenuadasRESUMO
Rotavirus A (RVA) vaccines succeeded in lowering the burden of acute gastroenteritis (AGE) worldwide, especially preventing severe disease and mortality. In 2019, Brazil completed 13 years of RVA vaccine implementation (Rotarix™) within the National Immunization Program (NIP), and as reported elsewhere, the use of Rotarix™ in the country has reduced childhood mortality and morbidity due to AGE. Even though both marketed vaccines are widely distributed, the surveillance of RVA causing AGE and the monitoring of circulating genotypes are important tools to keep tracking the epidemiological scenario and vaccines impact. Thus, our study investigated RVA epidemiological features, viral load and G and P genotypes circulation in children and adults presenting AGE symptoms in eleven states from three out of five regions in Brazil. By using TaqMan®-based one-step RT-qPCR, we investigated a total of 1536 stool samples collected from symptomatic inpatients, emergency department visits and outpatients from January 2018 to December 2019. G and P genotypes of RVA-positive samples were genetically characterized by multiplex RT-PCR or by nearly complete fragment sequencing. We detected RVA in 12% of samples, 10.5% in 2018 and 13.7% in 2019. A marked winter/spring seasonality was observed, especially in Southern Brazil. The most affected age group was children aged >24-60 months, with a positivity rate of 18.8% (p < 0.05). Evaluating shedding, we found a statistically lower RVA viral load in stool samples collected from children aged up to six months compared to the other age groups (p < 0.05). The genotype G3P[8] was the most prevalent during the two years (83.7% in 2018 and 65.5% in 2019), and nucleotide sequencing of some strains demonstrated that they belonged to the emergent equine-like G3P[8] genotype. The dominance of an emergent genotype causing AGE reinforces the need for continuous epidemiological surveillance to assess the impact of mass RVA immunization as well as to monitor the emergence of novel genotypes.
RESUMO
Brazil introduced rotavirus vaccination in March 2006. We studied 133 rotavirus-positive fecal samples collected from February 2005 through December 2007. Genotype G2P[4] was found in 1.4% of samples in 2005, in 44% in 2006, and in 96% in 2007. Rotavirus detection rate decreased from 38% in 2005 to 24% in 2007 (p = 0.012).
Assuntos
Fezes/virologia , Programas de Imunização , Avaliação de Programas e Projetos de Saúde , Infecções por Rotavirus , Rotavirus/genética , Rotavirus/isolamento & purificação , Brasil , Pré-Escolar , Diarreia/epidemiologia , Diarreia/prevenção & controle , Diarreia/virologia , Genótipo , Hospitalização , Humanos , Lactente , Recém-Nascido , Rotavirus/classificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/prevenção & controle , Infecções por Rotavirus/virologia , Vacinas contra Rotavirus/administração & dosagem , Vacinação/estatística & dados numéricosRESUMO
The Histo-blood group antigens (HBGA) are host genetic factors associated with susceptibility to rotavirus (RV) and human norovirus (HuNoV), the major etiological agents of viral acute gastroenteritis (AGE) worldwide. The FUT2 gene expressing the alpha-1, 2-L- fucosyltransferase enzyme is important for gut HBGA expression, and also provides a composition of the phenotypic profile achieved through mutations occurring in populations with different evolutionary histories; as such, it can be considered a genetic population marker. In this study, Lewis and secretor HBGA phenotyping was performed using 352 saliva samples collected from children between three months and five years old born in the Amazon (Brazil, Venezuela and English Guyana) presenting AGE or acute respiratory infection (ARI), the latter considered as control samples. The total of children phenotyped as secretors was 323, corresponding to 91.80%. From these, 207 (58.80%) had a Le (aâ¯+â¯b+) profile. The HBGA profiles were equally found in children with AGE as well as with ARI. The rs1047781 of the FUT2 gene was not detected in DNA from saliva cells with a Le (a+b+) profile. However, mutations not yet described in the FUT2 gene were observed: missense 325A>T, 501C>T, 585C>T, 855A>T and missense substitutions 327C>T [S (Ser) > C (Cys)], 446 T>C [L(Leu) > P(Pro)], 723C>A [N(Asn) > K(Lys)], 724A>T [I(Ile) > F(Phe)], 736C>A [H(His) > N(Asn)]. The SNP distribution in the FUT2 gene of the analyzed samples was very similar to that described in Asian populations, including indigenous tribes.
Assuntos
Infecções por Caliciviridae/epidemiologia , Fucosiltransferases/genética , Gastroenterite/epidemiologia , Predisposição Genética para Doença/etnologia , Antígenos do Grupo Sanguíneo de Lewis/genética , Infecções por Rotavirus/epidemiologia , Doença Aguda/epidemiologia , Brasil , Infecções por Caliciviridae/etnologia , Pré-Escolar , Feminino , Fucosiltransferases/sangue , Gastroenterite/virologia , Marcadores Genéticos , Humanos , Lactente , Antígenos do Grupo Sanguíneo de Lewis/sangue , Masculino , Polimorfismo de Nucleotídeo Único , Infecções Respiratórias , Infecções por Rotavirus/etnologia , Saliva/virologia , Venezuela , Galactosídeo 2-alfa-L-FucosiltransferaseRESUMO
Foodborne transmission gastroenteritis (AGE) outbreak occurred during a celebration lunch in July, 2016, Brazil. All stool samples tested were positive for noroviruses (NoV) and phylogenetic analysis revealed that strains were genetically close to GII.17 Kawasaki_2014. These findings indicated circulation of NoV GII.17 Kawasaki_2014 in the Brazilian population, associated with AGE outbreak.
Assuntos
Infecções por Caliciviridae/epidemiologia , Surtos de Doenças , Doenças Transmitidas por Alimentos/epidemiologia , Gastroenterite/epidemiologia , Norovirus/isolamento & purificação , Adulto , Brasil/epidemiologia , Infecções por Caliciviridae/transmissão , Infecções por Caliciviridae/virologia , Feminino , Doenças Transmitidas por Alimentos/virologia , Gastroenterite/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Norovirus/genética , Filogenia , Adulto JovemRESUMO
Group A rotaviruses (RVA) are one of the most common causes of severe acute gastroenteritis in infants worldwide. Rotaviruses spread from person to person, mainly by faecalâ»oral transmission. Almost all unvaccinated children may become infected with RVA in the first two years of life. The establishment of an experimental monkey model with RVA is important to evaluate new therapeutic approaches. In this study, we demonstrated viral shedding and viraemia in juvenileâ»adult Macaca fascicularis orally inoculated with Wa RVA prototype. Nine monkeys were inoculated orally: seven animals with human RVA and two control animals with saline solution. During the study, the monkeys were clinically monitored, and faeces and blood samples were tested for RVA infection. In general, the inoculated animals developed an oligosymptomatic infection pattern. The main clinical symptoms observed were diarrhoea in two monkeys for three days, associated with a reduction in plasmatic potassium content. Viral RNA was detected in seven faecal and five sera samples from inoculated animals, suggesting virus replication. Cynomolgus monkeys are susceptible hosts for human Wa RVA infection. When inoculated orally, they presented self-limited diarrhoea associated with presence of RVA infectious particles in faeces. Thus, cynomolgus monkeys may be useful as animal models to evaluate the efficacy of new antiviral approaches.
Assuntos
Infecções por Rotavirus/virologia , Rotavirus/fisiologia , Animais , Modelos Animais de Doenças , Fezes/virologia , Humanos , Macaca fascicularis , RNA Viral , Rotavirus/classificação , Infecções por Rotavirus/sangue , Carga Viral , Replicação Viral , Eliminação de Partículas ViraisRESUMO
Group A rotaviruses are the main cause of acute gastroenteritis in children throughout the world. The two outer capsid proteins, VP4 and VP7, define the P and G genotypes, respectively. Rotaviruses with P[8]G1, P[4]G2, P[8]G3 and P[8]G4 genotypes are predominant in infecting humans and the G9 genotype is emerging in most continents as the fifth most common G type worldwide. The inner capsid protein VP6 is responsible for subgroup (SG) specificities, allowing classification of rotaviruses into SG I, SG II, SG I+II and SG non-I-non-II. The non-structural protein 4 (NSP4) encoded by segment 10 has a role in viral morphogenesis and five genetic groups have been described, NSP4 genotypes A-E. The aim of this investigation was to characterize the NSP4 and VP6 genes of rotavirus strains recovered from hospitalized children. Thirty rotavirus strains were submitted to RT-PCR followed by sequencing and phylogenetic analysis. Among the different G and P genotype combinations, two distinct genetic groups could be recognized for the NSP4 gene. Twenty-eight clustered with NSP4 genotype B. The two P[4]G2 strains fell into NSP4 genotype A and clustered distinctly, with a 100 % bootstrap value. The strains distinguished within a group were closely related to each other at the nucleotide and amino acid levels. A phylogenetic tree was constructed for the VP6 gene including the human strains RMC100, E210, Wa, US1205 and 1076, and the animal strains Gott, NCDV, SA-11, FI-14 and EW. This is the first report on Brazilian rotavirus strains describing NSP4 genotype A strains associated with VP6 SG I, and NSP4 genotype B strains associated with VP6 SG II.
Assuntos
Antígenos Virais/genética , Proteínas do Capsídeo/genética , Glicoproteínas/genética , Infecções por Rotavirus/virologia , Rotavirus/classificação , Rotavirus/genética , Toxinas Biológicas/genética , Proteínas não Estruturais Virais/genética , Sequência de Aminoácidos , Brasil , Criança , Criança Hospitalizada , Análise por Conglomerados , Genótipo , Humanos , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rotavirus/isolamento & purificação , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
Epidemiological data on species A rotavirus (RVA) infections have demonstrated the genetic diversity of strains circulating worldwide. Many G and P genotype combinations have been described over the years, varying regionally and temporally, especially in developing countries. However, the most common G and P genotype combinations identified in RVA human strains worldwide are G1P[8], G2P[4], G3P[8], G4P[8] and G9P[8]. RVA genotype G1P[8] strains are responsible for more than 50% of child infections worldwide and component of the two vaccines (Rotarix® [RV1] and RotaTeq® [RV5]) licensed globally. For a better understanding of the evolutionary mechanisms of this genotype in Brazil, phylogenetic analyses based on the 11 RVA genome segments (genomic constellation) from 90 G1P[8] RVA strains collected in two eras - (i) pre-vaccination with RV1 (1996-February 2006); (ii) post-vaccination (March 2006-2013) - in different Brazilian states were performed. The results showed the Wa-like genomic constellation of the Brazilian G1P[8] strains with a I1-R1-C1-M1-A1-N1-T1-E1-H1 specificity, except for two strains (rj14055-07 and ba19030-10) that belong to a I1-R1-C1-M1-A1-N1-T3-E1-H1 genomic constellation, evidencing the occurrence of reassortment (Wa-like×AU-1-like) of the NSP3 gene. Reassortment events were also demonstrated between Brazilian G1P[8] strains and the RV1 vaccine strain in some genes in vaccinated and unvaccinated children. VP7 and VP8* antigenic site analysis showed that the amino acid substitutions observed in samples collected after the introduction of RV1 in Brazil were already detected in samples collected in the 1980s and 1990s, suggesting that mass Brazilian RV1 vaccination had no impact on the diversity observed inside antigenic sites for these two proteins.
Assuntos
Gastroenterite/virologia , Infecções por Rotavirus/virologia , Vacinas contra Rotavirus/genética , Rotavirus/genética , Vacinação/estatística & dados numéricos , Brasil/epidemiologia , Fezes/virologia , Gastroenterite/epidemiologia , Gastroenterite/prevenção & controle , Variação Genética/genética , Genoma Viral/genética , Genótipo , Humanos , Filogenia , RNA Viral/análise , RNA Viral/genética , Rotavirus/classificação , Rotavirus/imunologia , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/prevenção & controle , Vacinas contra Rotavirus/imunologia , Seleção Genética , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologiaRESUMO
Aichi viruses (AiV) have been detected in patients with diarrheal diseases (DD). The aim of this study was to assess AiV infection rates in hospitalized children with DD, including 123 HIV-1 seropositive and 125 HIV-1 seronegative patients, in two public pediatric hospitals in Rio de Janeiro, Brazil. AiV was investigated by nested RT-PCR. The AiV-positive samples were also tested for specie A rotavirus, norovirus, astrovirus, enteric adenovirus and bocavirus in order to assess co-infections. AiV parcial genome sequencing and phylogenetic analyses were performed. AiV were detected in 9/123 (7.32%) of the HIV-1 seropositive subjects and 1/125 (0.8%) of the HIV seronegative patients with DD (p = 0.019). The phylogenetic analysis of positive samples disclosed that: i) 13 samples were characterized as genotype A, with one of them being from the HIV-1 seronegative patient; ii) one sample from a HIV-1 seropositive patient was characterized as genotype B. AiV genotype A was grouped into 3 genetic clusters. Data suggest that AiV may be an opportunistic pathogen infecting children with AIDS and DD.
Assuntos
Infecções Oportunistas Relacionadas com a AIDS/virologia , Diarreia/virologia , Gastroenteropatias/virologia , Soropositividade para HIV/virologia , Kobuvirus/isolamento & purificação , Infecções por Picornaviridae/virologia , Brasil , Criança , Criança Hospitalizada , Pré-Escolar , Coinfecção/virologia , Fezes/virologia , Feminino , Soronegatividade para HIV , HIV-1 , Humanos , Lactente , Kobuvirus/genética , Masculino , FilogeniaRESUMO
This study aims to estimate the frequency of group A rotaviruses (RVA) infection with genotypes G3P[8] and G9P[8] in children that suffered from diarrheal disease (DD) between 2001 and 2011 in different Brazilian regions. In addition, the genetic diversity of G3P[8] and G9P[8] RVA strains recovered from vaccinated and non-vaccinated children was assessed. Laboratory-based RVA surveillance included 15,115 cases of DD, and RVA was detected by enzyme immune-assay and/or polyacrylamide gel electrophoresis in 3357 (22%) samples. RVA was genotyped by the semi-nested RT-PCR and among RVA-positive samples, 100 (2.9%) were G3 (63 G3P[8], 32 G3P not typed [NT], and 5 G3P[6]) and 378 (16.2%) were G9 (318 G9P[8], 59 G9P[NT], and 1 G9P[6]). From the G3 and G9 positive samples, 16 and 12, respectively, were obtained from children aged 4-48months vaccinated with the monovalent vaccine (Rotarix®, RV1). Phylogenetic analyses of the VP7 and VP8(∗) encoding genes were performed for 26 G3P[8] and 48 G9P[8] strains. VP8(∗) phylogenetic analysis revealed that all strains analyzed belonged to P[8] lineage III, whereas RV1 belongs to P[8]-I lineage. VP7 analysis revealed that all G3 and G9 strains belonged to G3-lineage III and G9-lineage III. The comparison of the VP7 and VP8(∗) antigenic epitopes regions of Brazilian strains with RV1 strain revealed several amino acid changes. However, no particular differences among Brazilian strains detected before and after vaccine introduction were observed, or among strains detected from vaccinated and non-vaccinated children. Complete genome characterization of four G3P[8] and seven G9P[8] strains revealed a typical conserved human Wa-like genomic constellation. Changes in the genetic diversity of G3P[8] and G9P[8] RVA detected from 2001 to 2011 in Brazil seemed not be related to RV1 introduction in Brazil.