Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 9.908
Filtrar
1.
J Med Microbiol ; 70(9)2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34516365

RESUMO

Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real-time PCR analysis.Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination.Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures.Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real-time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography.Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P <0.05), Salmonella spp. (P <0.05) and Campylobacter spp. (P <0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis.Conclusion. These results support the value of multiplex real-time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile micro-organisms. The identification of unsuspected micro-organisms for less specific clinical presentations may also impact on clinical practice and help optimize patient management.


Assuntos
Gastroenterite/diagnóstico , Reação em Cadeia da Polimerase Multiplex , Reação em Cadeia da Polimerase em Tempo Real , Adenoviridae/isolamento & purificação , Blastocystis hominis/isolamento & purificação , Campylobacter/isolamento & purificação , Cryptosporidium/isolamento & purificação , Dientamoeba/isolamento & purificação , Entamoeba histolytica/isolamento & purificação , Fezes/microbiologia , Fezes/parasitologia , Fezes/virologia , Gastroenterite/microbiologia , Gastroenterite/parasitologia , Giardia lamblia/isolamento & purificação , Humanos , Norovirus/isolamento & purificação , Rotavirus/isolamento & purificação , Salmonella/isolamento & purificação , Shigella/isolamento & purificação
2.
Water Res ; 204: 117615, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34492362

RESUMO

Faecal pathogens can be introduced into surface water through open defecation, illegal disposal and inadequate treatment of faecal sludge and wastewater. Despite sanitation improvements, poor countries are progressing slowly towards the United Nation's Sustainable Development Goal 6 by 2030. Sanitation-associated pathogenic contamination of surface waters impacted by future population growth, urbanization and climate change receive limited attention. Therefore, a model simulating human rotavirus river inputs and concentrations was developed combining population density, sanitation coverage, rotavirus incidence, wastewater treatment and environmental survival data, and applied to Uganda. Complementary surface runoff and river discharge data were used to produce spatially explicit rotavirus outputs for the year 2015 and for two scenarios in 2050. Urban open defecation contributed 87%, sewers 9% and illegal faecal sludge disposal 3% to the annual 15.6 log10 rotavirus river inputs in 2015. Monthly concentrations fell between -3.7 (Q5) and 2.6 (Q95) log10 particles per litre, with 1.0 and 2.0 median and mean log10 particles per litre, respectively. Spatially explicit outputs on 0.0833 × 0.0833° grids revealed hotspots as densely populated urban areas. Future population growth, urbanization and poor sanitation were stronger drivers of rotavirus concentrations in rivers than climate change. The model and scenario analysis can be applied to other locations.


Assuntos
Rotavirus , Qualidade da Água , Humanos , Rios , Uganda , Urbanização
3.
mBio ; 12(4): e0178121, 2021 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-34372695

RESUMO

The 2',5'-oligoadenylate (2-5A)-dependent endoribonuclease, RNase L, is a principal mediator of the interferon (IFN) antiviral response. Therefore, the regulation of cellular levels of 2-5A is a key point of control in antiviral innate immunity. Cellular 2-5A levels are determined by IFN-inducible 2',5'-oligoadenylate synthetases (OASs) and by enzymes that degrade 2-5A. Importantly, many coronaviruses (CoVs) and rotaviruses encode 2-5A-degrading enzymes, thereby antagonizing RNase L and its antiviral effects. A-kinase-anchoring protein 7 (AKAP7), a mammalian counterpart, could possibly limit tissue damage from excessive or prolonged RNase L activation during viral infections or from self-double-stranded RNAs that activate OAS. We show that these enzymes, members of the two-histidine phosphoesterase (2H-PE) superfamily, constitute a subfamily referred here as 2',5'-PEs. 2',5'-PEs from the mouse CoV mouse hepatitis virus (MHV) (NS2), Middle East respiratory syndrome coronavirus (MERS-CoV) (NS4b), group A rotavirus (VP3), and mouse (AKAP7) were investigated for their evolutionary relationships and activities. While there was no activity against 3',5'-oligoribonucleotides, they all cleaved 2',5'-oligoadenylates efficiently but with variable activity against other 2',5'-oligonucleotides. The 2',5'-PEs are shown to be metal ion-independent enzymes that cleave trimer 2-5A (2',5'-p3A3) producing mono- or diadenylates with 2',3'-cyclic phosphate termini. Our results suggest that the elimination of 2-5A might be the sole function of viral 2',5'-PEs, thereby promoting viral escape from innate immunity by preventing or limiting the activation of RNase L. IMPORTANCE Viruses often encode accessory proteins that antagonize the host antiviral immune response. Here, we probed the evolutionary relationships and biochemical activities of two-histidine phosphoesterases (2H-PEs) that allow some coronaviruses and rotaviruses to counteract antiviral innate immunity. In addition, we investigated the mammalian enzyme AKAP7, which has homology and shared activities with the viral enzymes and might reduce self-injury. These viral and host enzymes, which we refer to as 2',5'-PEs, specifically degrade 2',5'-oligoadenylate activators of the antiviral enzyme RNase L. We show that the host and viral enzymes are metal ion independent and exclusively cleave 2',5'- and not 3',5'-phosphodiester bonds, producing cleavage products with cyclic 2',3'-phosphate termini. Our study defines 2',5'-PEs as enzymes that share characteristic conserved features with the 2H-PE superfamily but have specific and distinct biochemical cleavage activities. These findings may eventually lead to pharmacological strategies for developing antiviral drugs against coronaviruses, rotaviruses, and other viruses.


Assuntos
Proteínas de Ancoragem à Quinase A/metabolismo , Nucleotídeos de Adenina/metabolismo , Endorribonucleases/metabolismo , Coronavírus da Síndrome Respiratória do Oriente Médio/enzimologia , Vírus da Hepatite Murina/enzimologia , Oligorribonucleotídeos/metabolismo , Rotavirus/enzimologia , Animais , Humanos , Imunidade Inata/imunologia , Interferons/imunologia , Camundongos
4.
Water Res ; 203: 117508, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34375933

RESUMO

Enteric viruses are commonly present in water bodies in regions with poor sanitation. Although the occurrence of these viruses poses a health risk they are difficult to quantify due to their low concentration and they may remain undetected in the absence of adequate preconcentration. The present study reports the synthesis and utilization of DEAE silica gel (DSiG) as an adsorbent for virus concentration. Two coliphages, MS2 and SUSP2, and an enteric virus, rotavirus A (RVA) were chosen for examining the preconcentration efficiency of DSiG columns. Studies conducted at a low flow rate of 5 mL/min yielded good removal of viruses through adsorption. Studies at a higher flow rate of 50 mL/min followed by elution with optimized eluents yielded a high recovery of MS2 and RVA even when they were present at low concentration (0.01 copy/mL). The eluent Na(1.5 M)-Tw(2%)-G3X (glycine 3X broth, 1.5 M NaCl, 2% Tween, pH 10.2) showed maximum elution of RVA and MS2. Optimal SUSP2 recovery was observed on employing an eluent composed of 1.5 M NaCl, 3% Tween, 0.05 M KH2PO4 at pH 9.2. Subsequently, both the eluents were successively applied for elution of the adsorbed viruses. This method was applied for virus preconcentration from lake water in the monsoon and winter seasons. The DSiG column could achieve adequate preconcentration for all the three viruses, i.e., SUSP2, MS2, and RVA, even when they were present at very low concentration and the recovery achieved was comparable to that achieved with ultracentrifugation while the processing time required for handling large volumes of water was considerably lower.


Assuntos
Rotavirus , Colífagos , Etanolaminas , Concentração de Íons de Hidrogênio , Sílica Gel , Água
5.
Viruses ; 13(7)2021 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-34372536

RESUMO

Equine rotavirus group A (ERVA) is one of the most common causes of foal diarrhea. Starting in February 2021, there was an increase in the frequency of severe watery to hemorrhagic diarrhea cases in neonatal foals in Central Kentucky. Diagnostic investigation of fecal samples failed to detect evidence of diarrhea-causing pathogens including ERVA. Based on Illumina-based metagenomic sequencing, we identified a novel equine rotavirus group B (ERVB) in fecal specimens from the affected foals in the absence of any other known enteric pathogens. Interestingly, the protein sequence of all 11 segments had greater than 96% identity with group B rotaviruses previously found in ruminants. Furthermore, phylogenetic analysis demonstrated clustering of the ERVB with group B rotaviruses of caprine and bovine strains from the USA. Subsequent analysis of 33 foal diarrheic samples by RT-qPCR identified 23 rotavirus B-positive cases (69.69%). These observations suggest that the ERVB originated from ruminants and was associated with outbreaks of neonatal foal diarrhea in the 2021 foaling season in Kentucky. Emergence of the ruminant-like group B rotavirus in foals clearly warrants further investigation due to the significant impact of the disease in neonatal foals and its economic impact on the equine industry.


Assuntos
Doenças dos Cavalos/virologia , Cavalos/virologia , Rotavirus/patogenicidade , Animais , Proteínas do Capsídeo/genética , Diarreia/etiologia , Diarreia/virologia , Surtos de Doenças/veterinária , Fezes/virologia , Kentucky , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Rotavirus/classificação , Infecções por Rotavirus/veterinária
6.
Viruses ; 13(7)2021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34372555

RESUMO

Viroplasms are cytoplasmic, membraneless structures assembled in rotavirus (RV)-infected cells, which are intricately involved in viral replication. Two virus-encoded, non-structural proteins, NSP2 and NSP5, are the main drivers of viroplasm formation. The structures (as far as is known) and functions of these proteins are described. Recent studies using plasmid-only-based reverse genetics have significantly contributed to elucidation of the crucial roles of these proteins in RV replication. Thus, it has been recognized that viroplasms resemble liquid-like protein-RNA condensates that may be formed via liquid-liquid phase separation (LLPS) of NSP2 and NSP5 at the early stages of infection. Interactions between the RNA chaperone NSP2 and the multivalent, intrinsically disordered protein NSP5 result in their condensation (protein droplet formation), which plays a central role in viroplasm assembly. These droplets may provide a unique molecular environment for the establishment of inter-molecular contacts between the RV (+)ssRNA transcripts, followed by their assortment and equimolar packaging. Future efforts to improve our understanding of RV replication and genome assortment in viroplasms should focus on their complex molecular composition, which changes dynamically throughout the RV replication cycle, to support distinct stages of virion assembly.


Assuntos
Rotavirus/genética , Rotavirus/metabolismo , Compartimentos de Replicação Viral/metabolismo , Animais , Proteínas do Capsídeo/genética , Citoplasma/virologia , Citosol/metabolismo , Humanos , Fosforilação , Proteínas de Ligação a RNA/metabolismo , Infecções por Rotavirus/virologia , Proteínas não Estruturais Virais/metabolismo , Compartimentos de Replicação Viral/fisiologia , Montagem de Vírus , Replicação Viral/genética
7.
Clin Infect Dis ; 73(4): e913-e920, 2021 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-34398953

RESUMO

BACKGROUND: Acute gastroenteritis (AGE) causes a substantial burden in the United States, but its etiology frequently remains undetermined. Active surveillance within an integrated healthcare delivery system was used to estimate the prevalence and incidence of medically attended norovirus, rotavirus, sapovirus, and astrovirus. METHODS: Active surveillance was conducted among all enrolled members of Kaiser Permanente Northwest during July 2014-June 2016. An age-stratified, representative sample of AGE-associated medical encounters were recruited to provide a stool specimen to be tested for norovirus, rotavirus, sapovirus, and astrovirus. Medically attended AGE (MAAGE) encounters for a patient occurring within 30 days were grouped into 1 episode, and all-cause MAAGE incidence was calculated. Pathogen- and healthcare setting-specific incidence estimates were calculated using age-stratified bootstrapping. RESULTS: The overall incidence of MAAGE was 40.6 episodes per 1000 person-years (PY), with most episodes requiring no more than outpatient care. Norovirus was the most frequently detected pathogen, with an incidence of 5.5 medically attended episodes per 1000 PY. Incidence of norovirus MAAGE was highest among children aged < 5 years (20.4 episodes per 1000 PY), followed by adults aged ≥ 65 years (4.5 episodes per 1000 PY). Other study pathogens showed similar patterns by age, but lower overall incidence (sapovirus: 2.4 per 1000 PY; astrovirus: 1.3 per 1000 PY; rotavirus: 0.5 per 1000 PY). CONCLUSIONS: Viral enteropathogens, particularly norovirus, are important contributors to MAAGE, especially among children < 5 years of age. The present findings underline the importance of judicious antibiotics use for pediatric AGE and suggest that an effective norovirus vaccine could substantially reduce MAAGE.


Assuntos
Infecções por Caliciviridae , Gastroenterite , Norovirus , Infecções por Rotavirus , Rotavirus , Sapovirus , Adulto , Infecções por Caliciviridae/epidemiologia , Criança , Fezes , Gastroenterite/epidemiologia , Humanos , Lactente , Estados Unidos/epidemiologia
8.
Int J Infect Dis ; 111: 76-84, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34411719

RESUMO

OBJECTIVE: To compare the intestinal microbiota profiles in infants following rotavirus (RV) and human norovirus (HNoV) infection. METHODS: Faecal specimens from 18 infants {mean age 11.8 months [standard deviation (SD) 3.0] months} with acute gastroenteritis caused by RV (G9P8) and 24 infants [mean age 8.8 (SD 6.4) months] with acute gastroenteritis caused by HNoV (GII) infection were collected prospectively. The faecal microbiome was assessed by 16S rRNA amplicon pyrosequencing. Alpha diversity, beta diversity, deferentially abundant taxa and microbial functions were assessed by bioinformatic analysis. RESULTS: The Chao1 index for the HNoV group was significantly higher compared with the control group (P=0.0003), and was lower for the RV group compared with the HNoV group (P=0.0078). No significant difference in beta diversity was observed between the RV and HNoV groups. The RV group showed greater abundance of Actinobacteria at phylum level and Bifidobacterium spp., Streptococcus spp., Enterococcus spp. and Lactobacillus spp. at genus level. The HNoV group showed richness in Fusobacteria and Cyanobacteria at phylum level, and Enterococcus spp. and Streptococcus spp. at genus level. Bacillus was the characteristic genus in infected infants. In comparison with the control group, the viral group (P≤0.01), the RV group (P=0.002) and the HNoV group (P≤0.01) showed significant differences in potentially pathogenic bacteria. CONCLUSIONS: Changes in microbiotic structure were observed in infants following RV and HNoV infection. The Chao 1 index of alpha diversity increased significantly in the HNoV group. Bacillus was the characteristic genus in infected infants. An increase in pathogenic bacteria, particularly Streptococcus spp. and Enterococcus spp., was detected in infected infants.


Assuntos
Infecções por Caliciviridae , Gastroenterite , Microbioma Gastrointestinal , Infecções por Rotavirus , Rotavirus , Criança , Fezes , Humanos , Lactente , Estudos Prospectivos , RNA Ribossômico 16S/genética , Rotavirus/genética
10.
Vaccine ; 39(33): 4577-4590, 2021 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-34244008

RESUMO

BACKGROUND: Older children and adults are susceptible to rotavirus, but the extent to which rotavirus affects this population is not fully understood, hindering accuracy of global rotavirus estimations. OBJECTIVE: To determine what proportion of diarrhea cases are due to rotavirus among persons ≥ 5 years old and to estimate this proportion by age strata. METHODS: We conducted a systematic review and meta-analysis using the PRISMA guidelines. We included studies that reported on conditional rotavirus prevalence (i.e., percent of diarrhea due to rotavirus) in persons ≥ 5 years old who were symptomatic with diarrhea/gastroenteritis and had laboratory confirmation for rotavirus infection. Studies on nosocomial infections and outbreak investigations were excluded. We collected age group-specific conditional rotavirus prevalence and other variables, such as study geography, study setting, and study type. We calculated pooled conditional rotavirus prevalence, corresponding 95% confidence intervals (95% CI), heterogeneity (I2) estimates, and prediction intervals (PI). RESULTS: Sixty-six studies from 32 countries met the inclusion criteria. Conditional rotavirus prevalence ranged from 0% to 30% across the studies. The total pooled prevalence of rotavirus among persons ≥ 5 years old with diarrhea was 7.6% (95% CI: 6.2-9.2%, I2 = 99.6%, PI: 0-24%). The pooled prevalence of rotavirus among older children and adolescents was 8.7% (95% CI: 6.2-11.7%, I2 = 96%, PI:0-27%), among younger adults was 5.4% (95% CI: 1.4-11.8%, I2 = 96%, PI:0-31%), and among older adults was 4.7% (95% CI: 2.8-7.0%, I2 = 96%, PI:0-16%). Pooled conditional rotavirus prevalences did not differ by other variables. CONCLUSION: In this systematic review and meta-analysis of rotavirus among persons ≥ 5 years old with diarrhea, we found relatively low pooled conditional rotavirus prevalence compared to what is typically reported for children < 5 years; however, results should be interpreted with caution as the wide prediction intervals suggest large heterogeneity.


Assuntos
Gastroenterite , Infecções por Rotavirus , Rotavirus , Adolescente , Idoso , Criança , Pré-Escolar , Diarreia/epidemiologia , Fezes , Humanos , Lactente , Prevalência , Infecções por Rotavirus/epidemiologia
11.
Vaccine ; 39(35): 4979-4987, 2021 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-34325930

RESUMO

BACKGROUND: Virus-like particles (VLPs) are unable to replicate in the recipient but stimulate the immune system through recognition of repetitive subunits. Parenterally delivered rotavirus-VLP (Ro-VLP) vaccine could have the potential to overcome the weaknesses of licensed oral live-attenuated rotavirus vaccines, namely, low efficacy in low-income and high mortality settings and a potential risk of intussusception. METHODS: A monovalent Ro-VLP composed of viral protein (VP) 7, VP6 and VP2 of G1 genotype specificity was produced in Nicotiana benthamiana using Agrobacterium tumefaciens infiltration-based transient recombinant expression system. Plants expressing recombinant G1 Ro-VLP were harvested, then the resultant biomass was processed through a series of clarification and purification steps including standard extraction, filtration, ultrafiltration and chromatography. The purified G1 Ro-VLP was subsequently examined for its immunogenicity and toxicological profile using animal models. RESULTS: G1 Ro-VLP had a purity of ≥90% and was structurally similar to triple-layered rotavirus particles as determined by cryogenic transmission electron microscopy. Two doses of aluminum hydroxide-adjuvanted G1 Ro-VLP (1 µg, 5 µg or 30 µg), administered intramuscularly, elicited a robust homotypic neutralizing antibody response in rats. Also, rabbits administered G1 Ro-VLP (10 µg or 30 µg) four times intramuscularly with aluminum hydroxide adjuvant did not show any significant toxicity. CONCLUSIONS: Plant-derived Ro-VLP composed of VP7, VP6 and VP2 structural proteins would be a plausible alternative to live-attenuated oral rotavirus vaccines currently distributed worldwide.


Assuntos
Infecções por Rotavirus , Vacinas contra Rotavirus , Rotavirus , Vacinas de Partículas Semelhantes a Vírus , Animais , Anticorpos Antivirais , Coelhos , Ratos , Rotavirus/genética , Infecções por Rotavirus/prevenção & controle , Vacinas Atenuadas , Vacinas de Partículas Semelhantes a Vírus/genética
12.
Virol J ; 18(1): 133, 2021 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-34193190

RESUMO

Investigation of human enterovirus (EV) in diarrheic fecal specimens is valuable to address EV diversity circulating worldwide. However, the detection of EV strains exclusively in fecal specimens must be interpreted cautiously. EV are well known causative agents associated with a spectrum of human diseases, but not acute gastroenteritis. EV isolation in stool samples could not necessarily be associated with diarrheic symptoms, as most EV infections appear to be asymptomatic, and healthy children could excrete EV in their stool. The diagnostic of EV is only confirmed when the neutralization test presents a significant increase in antibody titers (three times or more) in the paired serum samples (acute-phase and convalescent-phase) against the same EV serotype isolated in feces. In addition, patients suffering from acute gastroenteritis, even during an EV investigation, must be screened in parallel for gastroenteric viruses (i.e. norovirus and rotavirus) in order to clarify if the symptoms could be linked to other viral agent detected in their fecal samples. Surveillance of EV diversity among distinct patient groups, including diarrheic individuals, must be taken into consideration and can considerably increase the power of non-polio EV surveillance system in Brazil. More well-designed studies are necessary to further elucidate the role of EV in acute gastroenteritis.


Assuntos
Infecções por Enterovirus , Enterovirus , Gastroenterite , Rotavirus , Brasil , Criança , Infecções por Enterovirus/diagnóstico , Infecções por Enterovirus/epidemiologia , Fezes , Gastroenterite/diagnóstico , Gastroenterite/epidemiologia , Humanos
13.
Viruses ; 13(6)2021 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-34199978

RESUMO

Rotavirus A (RVA) has been considered the main cause of diarrheal disease in children under five years in emergency services in both developed and developing countries. RVA belongs to the Reoviridae family, which comprises 11 segments of double-stranded RNA (dsRNA) as a genomic constellation that encodes for six structural and five to six nonstructural proteins. RVA has been classified in a binary system with Gx[Px] based on the spike protein (VP4) and the major outer capsid glycoprotein (VP7), respectively. The emerging equine-like G3P[8] DS-1-like strains reported worldwide in humans have arisen an important concern. Here, we carry out the complete genome characterization of a previously reported G3P[8] strain in order to recognize the genetic diversity of RVA circulating among infants in Colombia. A near-full genome phylogenetic analysis was done, confirming the presence of the novel equine-like G3P[8] with a Wa-like backbone for the first time in Colombia. This study demonstrated the importance of surveillance of emerging viruses in the Colombian population; furthermore, additional studies must focus on the understanding of the spread and transmission dynamic of this important RVA strain in different areas of the country.


Assuntos
Diarreia/epidemiologia , Diarreia/virologia , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Rotavirus , Criança , Colômbia/epidemiologia , Doenças Transmissíveis Emergentes/diagnóstico , Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/virologia , Diarreia/diagnóstico , Genes Virais , Genoma Viral , Genômica , Genótipo , Humanos , Filogenia , Estudos Retrospectivos , Rotavirus/classificação , Rotavirus/genética , Infecções por Rotavirus/diagnóstico , Análise de Sequência de DNA
15.
Biosens Bioelectron ; 192: 113503, 2021 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-34303138

RESUMO

The COVID-19 pandemic has unfortunately demonstrated how easily infectious diseases can spread and harm human life and society. As of writing, pandemic has now been on-going for more than one year. There is an urgent need for new nucleic acid-based methods that can be used to diagnose pathogens early, quickly, and accurately to effectively impede the spread of infections and gain control of epidemics. We developed a flap probe-based isothermal nucleic acid amplification method that is triggered by recombinant FEN1-Bst DNA polymerase, which-through enzymatic engineering-has both DNA synthesis, strand displacement and cleavage functions. This novel method offers a simpler and more specific probe-primer pair than those of other isothermal amplifications. We tested the method's ability to detect SARS-CoV-2 (both ORF1ab and N genes), rotavirus, and Chlamydia trachomatis. The limits of detection were 10 copies/µL for rotavirus, C. trachomatis, and SARS-CoV-2 N gene, and 100 copies/µL for SARS-CoV-2 ORF1ab gene. There were no cross-reactions among 11 other common pathogens with characteristics similar to those of the test target, and the method showed 100% sensitivity and 100% specificity in clinical comparisons with RT-PCR testing. In addition to real-time detection, the endpoint could be displayed under a transilluminator, which is a convenient reporting method for point-of-care test settings. Therefore, this novel nucleic acid senor has great potential for use in clinical diagnostics, epidemic prevention, and epidemic control.


Assuntos
Técnicas Biossensoriais , Chlamydia trachomatis/isolamento & purificação , Rotavirus/isolamento & purificação , SARS-CoV-2/isolamento & purificação , COVID-19 , DNA Recombinante , DNA Polimerase Dirigida por DNA , Endonucleases Flap , Humanos , Técnicas de Amplificação de Ácido Nucleico , Pandemias , Sensibilidade e Especificidade
16.
Appl Environ Microbiol ; 87(18): e0071021, 2021 08 26.
Artigo em Inglês | MEDLINE | ID: mdl-34232739

RESUMO

In this study, the adsorption-elution method was modified to concentrate viral particles in water samples and investigate the contamination of groundwater with norovirus genogroup II (NoV GII), rotavirus A (RVA), and Pepper mild mottle virus (PMMoV). The mean recovery rate of a murine norovirus strain, which was inoculated into groundwater samples collected from a deep well, was the highest (39%) when the viral RNA was directly extracted from the membrane instead of eluting the adsorbed viral particles. This adsorption-direct extraction method was applied to groundwater samples (20 liters) collected from deep wells used for the public drinking water supply (n = 22) and private wells (n = 9). RVA (85 copies/liter) and NoV GII (35 copies/liter) were detected in water samples from a deep well and a private well, respectively. PMMoV was detected in 95% and 89% of water samples from deep wells and private wells, respectively, at concentrations of up to 990 copies/liter. The modified method was also used to extract bacterial DNA from the membrane (recovery rate of inoculated Escherichia coli K-12 was 22%). The Bacteroidales genetic markers specific to ruminants (BacR) and pigs (Pig2Bac) were detected in samples from a deep well and a private well, respectively. The modified virus concentration method has important implications for the management of microbiological safety in the groundwater supply. IMPORTANCE We investigated the presence of enteric viruses and bacterial genetic markers to determine fecal contamination in groundwater samples from deep wells used for the public drinking water supply and private wells in Japan. Groundwater is often subjected to chlorination; malfunctions in chlorine treatment result in waterborne disease outbreaks. The modified method successfully concentrated both viruses and bacteria in 20-liter groundwater samples. Norovirus genogroup II (GII), rotavirus A, Pepper mild mottle virus, and Bacteroidales genetic markers specific to ruminants and pigs were detected. Frequent flooding caused by increased incidences of extreme rainfall events promotes the infiltration of surface runoff containing livestock wastes and untreated wastewater into wells, possibly increasing groundwater contamination risk. The practical and efficient method developed in this study will enable waterworks and the environmental health departments of municipal/prefectural governments to monitor water quality. Additionally, the modified method will contribute to improving the microbiological safety of groundwater.


Assuntos
Água Potável/microbiologia , Água Subterrânea/microbiologia , Norovirus/genética , Rotavirus/genética , Tobamovirus/genética , Adsorção , Bacteroidetes/genética , DNA Bacteriano/genética , Monitoramento Ambiental , Escherichia coli K12/genética , Fezes/microbiologia , Genes Bacterianos , Japão , RNA Viral/genética , Microbiologia da Água , Poços de Água
17.
Acta Paediatr ; 110(10): 2833-2841, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34297362

RESUMO

AIM: To explore child health professionals' experiences of the early implementation of the rotavirus vaccination in the two regions that first introduced this vaccination in Sweden. METHODS: A descriptive and repeated cross-sectional study based on a digital study-specific questionnaire with a baseline in 2014 and with a 2-year follow-up in 2016. The study population consisted of nurses and doctors working in child health centres in the health care regions of Stockholm and Jönköping. RESULTS: In Stockholm, a larger proportion of the respondents (n = 355) had concerns in 2014, in comparison with the respondents in Jönköping (n = 101), mostly about the vaccination being a new and time-consuming task (60% versus 23%). In 2016, the overall attitude to vaccination was more positive in both regions and the levels of concern about increased workload were reduced (Stockholm, n = 519, 39%, versus Jönköping, n = 96, 10%). Challenges before and after the introduction in both regions were particularly related to how to give information about the vaccine's potential increased risk of intussusception. CONCLUSION: The gap between respondents' knowledge, attitudes and concerns pre- and post-vaccination introduction was larger in Stockholm compared to Jönköping. In both regions, overall, the implementation of the rotavirus vaccination was perceived as being easier than expected.


Assuntos
Rotavirus , Criança , Saúde da Criança , Estudos Transversais , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Suécia , Vacinação
18.
Arch Virol ; 166(9): 2369-2386, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34216267

RESUMO

Rotaviruses are segmented double-stranded RNA viruses with a high frequency of gene reassortment, and they are a leading cause of global diarrheal deaths in children less than 5 years old. Two-thirds of rotavirus-associated deaths occur in low-income countries. Currently, the available vaccines in developing countries have lower efficacy in children than those in developed countries. Due to added safety concerns and the high cost of current vaccines, there is a need to develop cost-effective next-generation vaccines with improved safety and efficacy. The reverse genetics system (RGS) is a powerful tool for investigating viral protein functions and developing novel vaccines. Recently, an entirely plasmid-based RGS has been developed for several rotaviruses, and this technological advancement has significantly facilitated novel rotavirus research. Here, we review the recently developed RGS platform and discuss its application in studying infection biology, gene reassortment, and development of vaccines against rotavirus disease.


Assuntos
Genética Reversa , Infecções por Rotavirus/prevenção & controle , Vacinas contra Rotavirus/genética , Rotavirus/genética , Animais , Pré-Escolar , Análise Custo-Benefício , Diarreia/prevenção & controle , Diarreia/virologia , Interações Hospedeiro-Patógeno , Humanos , Plasmídeos , RNA Viral/genética , Vacinas contra Rotavirus/economia , Proteínas Virais/genética
19.
Arch Virol ; 166(9): 2479-2486, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34218319

RESUMO

Rotaviruses are important causative agents of acute gastroenteritis in children. In China, rotavirus infection has a prevalence rate of 30% and is therefore considered a serious public health problem. This study was carried out to investigate the clinical and molecular epidemiological characteristics of rotavirus infections in children under 5 years old with acute diarrhea in Shandong province, China. From July 2017 to June 2018, a total of 1211 fecal specimens were tested, and the prevalence of rotavirus infection was 32.12%. The mean age of the infected children was 12.2 ± 10.9 months, and the highest infection rate was observed in children aged 7-12 months, with a rate of 41.64%. G9P[8] (76.61%) was the most prevalent genotype combination, followed by G2P[4] (7.20%), G3P[8] (3.60%), and G9P[4] (2.06%). In addition to diarrhea, vomiting, fever, and dehydration were the most common clinical signs. In general, there was no significant difference in clinical manifestations among different age groups. However, the clinical manifestations differed significantly between vaccinated and unvaccinated children. Vaccinated children showed lower incidence and frequency of vomiting, lower incidence and degree of dehydration, and lower incidence of severe cases than unvaccinated children. These findings suggest that it is necessary to continuously monitor changes in the characteristics of rotavirus infections. Moreover, the introduction of vaccines into the national immunization program to prevent and control rotavirus infection is needed in China.


Assuntos
Epidemiologia Molecular , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Rotavirus/genética , Pré-Escolar , China/epidemiologia , Diarreia/virologia , Feminino , Gastroenterite/virologia , Genótipo , Humanos , Programas de Imunização , Incidência , Lactente , Masculino , Prevalência , Infecções por Rotavirus/fisiopatologia , Infecções por Rotavirus/prevenção & controle , Vacinas contra Rotavirus , Vacinação
20.
Vaccine ; 39(32): 4463-4470, 2021 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-34218961

RESUMO

BACKGROUND AND AIMS: ROTAVIN-M1® (licensed, frozen vaccine) and ROTAVIN (second-generation, liquid candidate vaccine) are two rotavirus vaccine formulations developed from a live attenuated G1P8 (KH0118) strain by Center for Research and Production of Vaccines and Biologicals (POLYVAC), Vietnam. This study compared the safety and immunogenicity of these two formulations. METHODS: A Phase 3, randomized, partially double-blinded, active-controlled study was conducted in healthy infants aged 60-91 days in Vietnam. Infants received two doses of ROTAVIN or ROTAVIN-M1 in a ratio of 2:1 with an interval of 8 weeks. Solicited reactions were collected for 7 days after each vaccination. Blood samples were collected pre-vaccination and 4 weeks after the second vaccination in a subset of infants. Non-inferiority criteria required that the lower bound of 95% confidence intervals (CIs) of the post-vaccination anti-rotavirus IgA GMC (Geometric Mean Concentration) ratio of ROTAVIN/ROTAVIN-M1 should be >0.5. A co-primary objective was to compare the safety of the two vaccines in terms of solicited reactions. RESULTS: A total of 825 infants were enrolled. The post-vaccination GMC was 48.25 (95% CI: 40.59, 57.37) in the ROTAVIN group and 35.04 (95% CI: 27.34, 44.91) in the ROTAVIN-M1 group with an IgA GMC ratio of 1.38 (95% CI: 1.02, 1.86) thus meeting the pre-set criteria for non-inferiority. A total of 605 solicited reactions were reported in 297 (36.0%) participants with 35.4% in the ROTAVIN group and 37.2% in the ROTAVIN-M1 group. There were no cases of intussusception or death reported in the study. CONCLUSIONS: Based on the data generated, it can be concluded that ROTAVIN is immunologically non-inferior and has similar safety profile to ROTAVIN-M1 when administered to infants in a two-dose schedule. Therefore, it can be considered as a more suitable option for programmatic use to prevent rotavirus diarrhoea in Vietnam and the Mekong region. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov identifier: NCT03703336, October 11, 2018.


Assuntos
Infecções por Rotavirus , Vacinas contra Rotavirus , Rotavirus , Anticorpos Antivirais , Grupo com Ancestrais do Continente Asiático , Humanos , Imunogenicidade da Vacina , Lactente , Infecções por Rotavirus/prevenção & controle , Vacinas contra Rotavirus/efeitos adversos , Vacinas Atenuadas/efeitos adversos , Vietnã
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...