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1.
Zhonghua Liu Xing Bing Xue Za Zhi ; 40(8): 889-894, 2019 Aug 10.
Artigo em Chinês | MEDLINE | ID: mdl-31484249

RESUMO

Objective: To understand the seasonality and etiological characteristics of infectious diarrhea in adults from Shanghai. Methods: Adult patients with diarrhea who had visited the enteric disease clinics in 22 hospitals that carrying on the Diarrhea Comprehensive Surveillance sentinel programs in Shanghai during 2014-2017, were surveyed. Stool specimens were collected according to the different intervals of sampling and detected for 12 bacteria and 5 viruses. Concentration ratio and circular distribution method were used for data analysis. Results: From 2014 to 2017, a total of 9 573 stool specimens were collected from the targeted diarrhea patients ≥18 years old (n=96 067), through the Shanghai Diarrhea Comprehensive Surveillance program. The positive rate of detection was 46.44%. Seasonal peaks of infectious diarrhea were both seen in summer (bacteria peak, diarrheagenic Escherichia coli and Vibrio parahaemolyticus, etc.) and in winter (virus peak, Norovirus, etc.). Both bacterial and viral infections presented seasonal concentration (Raleigh's test P<0.001) but more obvious with bacterial infection. Viral infection accounted for 60.19% of the cause of infectious diarrhea. The top five predominant pathogens appeared as Norovirus, Rotavirus, diarrheagenic Escherichia coli, Vibrio parahaemolyticus, and Salmonella spp.. Conclusions: Among the adult outpatients with infectious diarrhea in Shanghai, obvious seasonality was seen, with peaks in both summer and winter. Viral infection with Norovirus in particular, appeared as the predominant source of infection. Active, continuous and comprehensive diarrhea-related surveillance programs would be able to monitor the changing dynamic of pathogen spectrum, and lead to the adoption of targeted preventive measures.


Assuntos
Bactérias/isolamento & purificação , Diarreia/diagnóstico , Diarreia/etiologia , Disenteria/diagnóstico , Disenteria/etiologia , Fezes , Pacientes Ambulatoriais/estatística & dados numéricos , Vigilância da População/métodos , Vírus/isolamento & purificação , Adolescente , Adulto , Bactérias/classificação , Criança , Pré-Escolar , China/epidemiologia , Diarreia/epidemiologia , Disenteria/epidemiologia , Escherichia coli/isolamento & purificação , Fezes/microbiologia , Fezes/virologia , Humanos , Pessoa de Meia-Idade , Norovirus/isolamento & purificação , Rotavirus/isolamento & purificação , Salmonella/classificação , Salmonella/isolamento & purificação , Estações do Ano , Vibrio parahaemolyticus/isolamento & purificação , Vírus/classificação
2.
Zhonghua Liu Xing Bing Xue Za Zhi ; 40(8): 895-899, 2019 Aug 10.
Artigo em Chinês | MEDLINE | ID: mdl-31484250

RESUMO

Objective: To conduct a viral pathogen surveillance program on pediatric inpatients less than five years old with acute gastroenteritis in Shanghai and to better understand the pathogenic spectrum and molecular features in the target population, for setting up programs on control, prevention, medication and vaccine applications of the diseases. Methods: Fecal samples were collected from inpatients less than 5 years old who were admitted to a pediatric hospital for having acute gastroenteritis. Information related to demographic, clinical and epidemiological features of the patients was also collected. Laboratory assays including ELISA, real-time PCR and nested PCR, were performed to detect the presence of pathogens as rotavirus, calicivirus, astrovirus and adenovirus. Results: A total of 1 018 samples were collected (male 671 and 347 female), with the positive detection rate as 40.57% which peaked from autumn till winter, annually. Calicivirus and rotavirus A presented with the highest detection rates (24.75% and 13.95% respectively). The lowest detection rate was found in the 0-6 month-olds (32.20%). 65% of the patients with positive virus had received antibiotic treatment prior to the hospitalization. However, no statistically significant difference was seen, regarding the rates of antibiotic medication in the virus positive or negative populations (P>0.05). Data from the Rotavirus genotype analysis revealed that G9P[8] genotype was the predominant strain, and causing majority of rotavirus infections in all the age groups. Conclusions: Among the inpatients under 5 years of age in Shanghai, the positive detection rate for Calicivirus was higher than that for rotavirus group A, suggesting the necessity to carefully monitor the changes regarding the pathogenic spectrum and subtypes of the virus. Antibiotics should also be attentively administered, together with the development of suitable vaccine.


Assuntos
Caliciviridae/isolamento & purificação , Fezes/virologia , Gastroenterite/virologia , Pacientes Internados/estatística & dados numéricos , Infecções por Rotavirus/diagnóstico , Rotavirus/isolamento & purificação , Doença Aguda , Pré-Escolar , China/epidemiologia , Feminino , Gastroenterite/diagnóstico , Gastroenterite/epidemiologia , Humanos , Lactente , Masculino , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia
3.
Pan Afr Med J ; 32: 202, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31312314

RESUMO

Introduction: Rotavirus causes severe-diarrheal diseases in infants. An estimation of 138 million rotavirus-associated diarrheal cases and 215,000 deaths occur every year globally. In December 2016, West-Shewa zone in Ethiopia reported unidentified gastrointestinal diarrhea outbreak. We investigated to identify the causative agent of the outbreak to support response operations. Methods: Medical records were reviewed, and the daily line list was collected from health facilities. Descriptive data analysis was done by time, person and place. Stool specimens were first tested by antigen capture enzyme immunoassay (EIA) technique and further confirmed by reverse-transcription polymerase chain reaction (RT-PCR) as a gold standard. The product of RT-PCR was genotyped for each gene using G1-G4, G8-G9 and G12 primers for VP7 gene and P(4), P(6), P(8) and P(14) primers for VP4 gene. Results: A total of 1,987 diarrheal cases (5.7 per 1000) and five deaths (case-fatality rate 0.25%) were identified and epidemiologically-linked to confirmed rotavirus from December 2016 to February 2017. Among the cases, 1,946 (98%) were < 5 children. Fourteen (74%) of the 19 tested stool specimens were positive for rotavirus by EIA and RT-PCR. Majority of strains detected were G12P(6) (25%) and G-negative P(8) (25%) followed by G9P(8) (19%), G1P(8) (13%) and G3/G2 P(8), G12P(8), and G-negative P(6) (6% each). Conclusion: Diarrheal outbreak which occurred in West-Shewa zone of Ethiopia was associated with rotavirus and relatively more affected districts with low vaccination coverage. Routine rotavirus vaccination quality and coverage should be evaluated and the surveillance system needs to be strengthened to detect, prevent and control a similar outbreak.


Assuntos
Diarreia/epidemiologia , Infecções por Rotavirus/epidemiologia , Vacinas contra Rotavirus/administração & dosagem , Rotavirus/isolamento & purificação , Doença Aguda , Adolescente , Adulto , Criança , Pré-Escolar , Diarreia/virologia , Surtos de Doenças , Etiópia/epidemiologia , Fezes/virologia , Feminino , Genótipo , Humanos , Técnicas Imunoenzimáticas/métodos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rotavirus/genética , Infecções por Rotavirus/virologia , Cobertura Vacinal/estatística & dados numéricos , Adulto Jovem
4.
MMWR Morb Mortal Wkly Rep ; 68(24): 539-543, 2019 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-31220058

RESUMO

Before the introduction of rotavirus vaccine in the United States in 2006, rotavirus infection was the leading cause of severe gastroenteritis among U.S. children (1). To evaluate the long-term impact of rotavirus vaccination on disease prevalence and seasonality in the United States, CDC analyzed national laboratory testing data for rotavirus from laboratories participating in CDC's National Respiratory and Enteric Viruses Surveillance System (NREVSS) during the prevaccine (2000-2006) and postvaccine (2007-2018) periods. Nationally, the median annual percentage of tests positive for rotavirus declined from 25.6% (range = 25.2-29.4) in the prevaccine period to 6.1% (range = 2.6-11.1) in the postvaccine period. When compared with the prevaccine period, the postvaccine period saw declines in the annual peak in rotavirus positivity from a median of 43.1% (range = 43.8-56.3) to a median of 14.0% (range = 4.8-27.3) and in the season duration from a median of 26 weeks (range = 23-27) to a median of 9 weeks (range = 0-18). In the postvaccine period, a biennial pattern emerged, with alternating years of low and high rotavirus activity. Implementation of the rotavirus vaccination program has substantially reduced prevalence of the disease and altered seasonal patterns of rotavirus in the United States; these changes have been sustained over 11 seasons after vaccine introduction. Ongoing efforts to improve coverage and on-time vaccination (2) can help maximize the public health impact of rotavirus vaccination.


Assuntos
Programas de Imunização/organização & administração , Laboratórios/tendências , Vigilância da População , Infecções por Rotavirus/diagnóstico , Infecções por Rotavirus/prevenção & controle , Vacinas contra Rotavirus/administração & dosagem , Rotavirus/isolamento & purificação , Criança , Humanos , Lactente , Avaliação de Programas e Projetos de Saúde , Infecções por Rotavirus/epidemiologia , Estações do Ano , Estados Unidos/epidemiologia
5.
J Med Microbiol ; 68(8): 1233-1239, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31215861

RESUMO

PURPOSE: Human bocavirus (HBoV) is a DNA virus that is mostly associated with respiratory infections. However, because it has been found in stool samples, it has been suggested that it may be a causative agent for human enteric conditions. This underpins the continuous search for HBoVs, especially after the introduction of the rotavirus vaccine due to acute gastroenteritis cases related to emergent viruses, as HBoVs are more likely to be found in this post-vaccine scenario. Therefore, the aim of this study is to demonstrate the prevalence of HBoV in children aged less than 10 years with acute gastroenteritis in Brazil from November 2011 to November 2012. METHODOLOGY: Stool samples from hospitalized children ≤10 years old who presented symptoms of acute gastroenteritis were analysed for the presence of rotavirus A (RVA) by an enzyme-linked immunosorbent assay (ELISA), and for HBoV DNA by nested PCR. RESULTS: HBoV positivity was detected in 24.0 % (54/225) of samples. Two peaks of HBoV detection were observed in November 2011 and from July to September 2012. Co-infections between HBoV and rotavirus A were identified in 50.0 % (27/54) of specimens. Phylogenetic analysis identified the presence of HBoV-1 (94.8 %), HBoV-2 (2.6 %) and HBoV-3 (2.6 %) species, with only minor variations among them. CONCLUSION: Our findings provide evidence for the circulation of most HBoV genotypes (except HBoV-4) in the North Region of Brazil at a considerable rate and further investigations are necessary to improve our knowledge in the context of HBoV infections and their role in gastrointestinal diseases.


Assuntos
Gastroenterite/epidemiologia , Bocavirus Humano/genética , Epidemiologia Molecular , Infecções por Parvoviridae/epidemiologia , Doença Aguda , Brasil/epidemiologia , Proteínas do Capsídeo/genética , Criança , Pré-Escolar , Coinfecção/epidemiologia , Coinfecção/virologia , DNA Viral/genética , Fezes/virologia , Feminino , Gastroenterite/virologia , Genótipo , Bocavirus Humano/classificação , Bocavirus Humano/isolamento & purificação , Humanos , Lactente , Recém-Nascido , Masculino , Infecções por Parvoviridae/virologia , Filogenia , Reação em Cadeia da Polimerase , Prevalência , Rotavirus/imunologia , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Análise de Sequência de DNA
6.
J Med Microbiol ; 68(8): 1240-1243, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31237533

RESUMO

The aim of the present study was to report the molecular characterization of human group A rotaviruses (RVAs) circulating in Tunisia. Stool specimens were collected from children under 5 years of age who had been hospitalized or were consulting for gastroenteritis in Tunisian hospitals between 2015 and 2017. All samples were screened by reverse-transcription polymerase chain reaction (RT-PCR) for the detection of the VP6 gene specific for RVA. RVA-positive samples were further analysed for G/P genotyping by semi-nested multiplex RT-PCR. Among 454 tested samples, 72 (15.8 %) were positive for RVA. G1P[8] was the most prevalent detected strain (41.7%), followed by G9P[8] (32.8%), G2P[4] (7.5%), G12P[8] (7.5%), G1P[6] (3.0%), G2P[8] (1.5%) and G3P[8] (1.5%), with mixed infections in 4.5 % of cases. In the absence of a national anti-rotavirus vaccination strategy, RVAs remain the primary aetiological agent for gastroenteritis in Tunisian children.


Assuntos
Gastroenterite/virologia , Infecções por Rotavirus/virologia , Rotavirus/genética , Proteínas do Capsídeo/genética , Pré-Escolar , Fezes/virologia , Gastroenterite/epidemiologia , Variação Genética , Genótipo , Humanos , Lactente , Recém-Nascido , Epidemiologia Molecular , Prevalência , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Estações do Ano , Análise de Sequência de DNA , Tunísia/epidemiologia
7.
Virol J ; 16(1): 64, 2019 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-31092258

RESUMO

BACKGROUND: Acute gastroenteritis (AGE) due to group A rotavirus (RVA) agent is one of the major causes of hospitalization in paediatric age. The G3P[8] RVA genotype has been usually considered as one of the major human genotypes, largely circulating in Asia, but showing low detection rates in the European countries. In recent years, the G3P[8] RVAs emerged also in Europe as a predominant genotype and the viral strains detected revealed high similarities with equine-like G3P[8] RVA strains, resulting in a new variant circulating in humans and able to cause AGE in the paediatric population. CASE PRESENTATION: An 8-year-old boy was admitted to the Emergency Room because he had suffered from severe diarrhoea, vomiting, and high fever over the previous two days. Severe dehydration was evident based on low serum concentrations of potassium and sodium, low glycaemia, and pre-renal failure (creatinine 2.48 mg/dL, urea 133 mg/dL). Immunological tests were within normal range. Enzyme immunoassay for the detection of RV was positive, and a sample of faeces was collected in order to perform the molecular characterization of the viral strain. The phylogenetic trees revealed relatedness between the VP7 and VP4 genes of the G3P[8] RVA Italian strain (namely PG2) and those belonging to recent G3P[8] RVAs detected worldwide. The G3 VP7 belonged to the G3-I lineage and shared the highest nucleotide sequence identity (99.8%) with the equine-like G3 previously identified in other countries. The P [8] VP4 revealed a similar clustering pattern to that observed for the VP7. In addition, the molecular characterization of the 11 gene segments of strain PG2 revealed a G3-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2 genomic constellation. CONCLUSIONS: This case shows the first detection in Italy of a reassortant G3P[8] RVA associated with a severe AGE, which is unusual in a school-age child without any known severe underlying problems. The findings reported in this paper highlight the importance of continuously monitoring the RVA strains circulating in paediatric age in order to detect novel viral variants able to spread in the general population.


Assuntos
Gastroenterite/virologia , Genótipo , Vírus Reordenados/genética , Infecções por Rotavirus/diagnóstico , Rotavirus/genética , Criança , Diarreia/virologia , Fezes/virologia , Gastroenterite/diagnóstico , Gastroenterite/terapia , Genoma Viral , Humanos , Infusões Intravenosas , Itália , Masculino , Vírus Reordenados/isolamento & purificação , Rotavirus/isolamento & purificação , Infecções por Rotavirus/terapia , Análise de Sequência de DNA
8.
Arch Virol ; 164(7): 1781-1791, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31079214

RESUMO

Introduction of animal group A rotavirus (RVA) gene segments into the human RVA population is a major factor shaping the genetic landscape of human RVA strains. The VP6 and NSP4 genes of 74 G/P-genotyped RVA isolates collected in Rawalpindi during 2010 were analyzed, revealing the presence of VP6 genotypes I1 (60.8%) and I2 (39.2%) and NSP4 genotypes E1 (60.8%), E2 (28.3%) and E-untypable (10.8%) among the circulating human RVA strains. The typical human RVA combinations I1E1 and I2E2 were found in 59.4% and 24.3% of the cases, respectively, whereas 5.4% of the RVA strains were reassortants, i.e., either I1E2 or I2E1. The phylogeny of the NSP4 gene showed that one G2P[4] and two G1P[6] RVA strains clustered with porcine E1 RVA strains or RVA strains that were considered to be (partially) of porcine origin. In addition, the NSP4 gene segment of the unusual human G6P[1] RVA strains clustered closely with bovine E2 RVA strains, further strengthening the hypothesis of an interspecies transmission event. The study further demonstrates the role of genomic re-assortment and the involvement of interspecies transmission in the evolution of human RVA strains. The VP6 and NSP4 nucleotide sequences analyzed in the study received the GenBank accession numbers KC846908- KC846971 and KC846972-KC847037, respectively.


Assuntos
Antígenos Virais/genética , Proteínas do Capsídeo/genética , Glicoproteínas/genética , Infecções por Rotavirus/transmissão , Rotavirus/genética , Rotavirus/isolamento & purificação , Toxinas Biológicas/genética , Proteínas não Estruturais Virais/genética , Sequência de Aminoácidos , Animais , Bovinos , Pré-Escolar , Gastroenterite/virologia , Genoma Viral/genética , Genótipo , Humanos , Paquistão , Filogenia , Infecções por Rotavirus/virologia , Alinhamento de Sequência , Suínos , Zoonoses/virologia
9.
Vet Microbiol ; 232: 105-113, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31030833

RESUMO

Rotaviruses of group A (RVAs) commonly occur in farm animals. In pigs, they cause acute gastrointestinal disease which is considered as significant factor of economic losses in pig farming. The aim of the study was an assessment of the prevalence of rotavirus (RV) infections in farmed pigs in Poland, genotype identification of the virus strains in conjunction with their age-related occurrence and regional (province) distribution pattern in pig herds. In total, 920 pig faecal samples were collected from pigs between the ages of one week and two years old from 131 farms. RVAs were detected using ELISA and molecular methods followed by a sequence-based identification of G (VP7) and P (VP4) virus genotypes. RV antigen was found in 377 (41%) of pig faecal samples. The correlation between pig age and frequency of RV infections was shown. In the Polish pig population, 145 RVA strains representing 33 GP genotypes were identified. Subsequent molecular analysis revealed an age-dependent and regional diversity in distribution of genotypes and virus strains. Besides typical pig RVA strains, novel strains such as G5P [34], G9P[34], and human G1P[8] were identified in this animal host. Findings from this study showed a change over time in the genotype occurrence of circulating pig RVAs in Poland. The high genetic variability of RV strains and acquisition of new virus genotypes have led to the emergence of novel, genetically distinct RVAs. The changes in the genotype occurrence of RVA strains in pigs indicate the need for their continuous epidemiological surveillance.


Assuntos
Variação Genética , Genótipo , Infecções por Rotavirus/veterinária , Rotavirus/classificação , Doenças dos Suínos/epidemiologia , Fatores Etários , Animais , Diarreia/virologia , Fezes/virologia , Genoma Viral , Filogenia , Polônia/epidemiologia , Prevalência , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Análise de Sequência de DNA , Suínos/virologia , Doenças dos Suínos/virologia
10.
Virol J ; 16(1): 49, 2019 04 25.
Artigo em Inglês | MEDLINE | ID: mdl-31023319

RESUMO

BACKGROUND: Equine rotavirus A (ERVA) is the leading cause of diarrhea in neonatal foals and has a negative impact on equine breeding enterprises worldwide. Among ERVA strains infecting foals, the genotypes G3P[12] and G14P[12] are the most prevalent, while infections by strains with other genomic arrangements are infrequent. The identification of circulating strains of ERVA is critical for diagnostic and surveillance purposes, as well as to understand their molecular epidemiology. Current genotyping methods available for ERVA and rotaviruses affecting other animal species rely on Sanger sequencing and are significantly time-consuming, costly and labor intensive. Here, we developed the first one-step multiplex TaqMan® real-time reverse transcription polymerase chain reaction (RT-qPCR) assay targeting the NSP3 and VP7 genes of ERVA G3 and G14 genotypes for the rapid detection and G-typing directly from fecal specimens. METHODS: A one-step multiplex TaqMan® RT-qPCR assay targeting the NSP3 and VP7 genes of ERVA G3 and G14 genotypes was designed. The analytical sensitivity was assessed using serial dilutions of in vitro transcribed RNA containing the target sequences while the analytical specificity was determined using RNA and DNA derived from a panel of group A rotaviruses along with other equine viruses and bacteria. The clinical performance of this multiplex assay was evaluated using a panel of 177 fecal samples and compared to a VP7-specific standard RT-PCR assay and Sanger sequencing. Limits of detection (LOD), sensitivity, specificity, and agreement were determined. RESULTS: The multiplex G3 and G14 VP7 assays demonstrated high specificity and efficiency, with perfect linearity. A 100-fold difference in their analytical sensitivity was observed when compared to the singleplex assays; however, this difference did not have an impact on the clinical performance. Clinical performance of the multiplex RT-qPCR assay demonstrated that this assay had a high sensitivity/specificity for every target (100% for NSP3, > 90% for G3 VP7 and > 99% for G14 VP7, respectively) and high overall agreement (> 98%) compared to conventional RT-PCR and sequencing. CONCLUSIONS: This new multiplex RT-qPCR assay constitutes a useful, very reliable tool that could significantly aid in the rapid detection and G-typing of ERVA strains circulating in the field.


Assuntos
Fezes/virologia , Técnicas de Genotipagem , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções por Rotavirus/veterinária , Rotavirus/genética , Rotavirus/isolamento & purificação , Animais , Antígenos Virais/genética , Proteínas do Capsídeo/genética , Diarreia/virologia , Genoma Viral , Genótipo , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/virologia , Cavalos , Reação em Cadeia da Polimerase Multiplex/normas , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Infecções por Rotavirus/diagnóstico , Sensibilidade e Especificidade , Proteínas não Estruturais Virais/genética
11.
Arch Virol ; 164(6): 1515-1525, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30887229

RESUMO

Rotaviruses and noroviruses are the most important viral causes of acute gastroenteritis in children. While previous studies of acute gastroenteritis in Indonesia mainly focused on rotavirus, here, we investigated the burden and epidemiology of norovirus and rotavirus disease. Children less than five years of age hospitalized with acute gastroenteritis were enrolled in this study from January to December 2015 at three participating hospitals. Rotavirus was detected by enzyme immunoassay (EIA), followed by genotyping by reverse transcription PCR (RT-PCR). Norovirus genogroups were determined by TaqMan-based quantitative RT-PCR. Among 406 enrolled children, 75 (18.47%), 223 (54.93%) and 29 (7.14%) cases were positive for norovirus, rotavirus and both viruses (mixed infections), respectively. Most cases clinically presented with fever, diarrhea, vomiting and some degree of dehydration. The majority (n = 69/75 [92%]) of the noroviruses identified belonged to genogroup II, and several genotypes were identified by sequencing a subset of samples. Among 35 samples tested for rotavirus genotype, the most prevalent genotype was G3P[8] (n = 30/35 [85.6%]). Our study suggests that the burden of norovirus diseases in Indonesian children should not be underestimated. It also shows the emergence of rotavirus genotype G3P[8] in Indonesia.


Assuntos
Infecções por Caliciviridae/diagnóstico , Gastroenterite/virologia , Norovirus/classificação , Infecções por Rotavirus/diagnóstico , Rotavirus/classificação , Pré-Escolar , Fezes/virologia , Feminino , Técnicas de Genotipagem/métodos , Hospitalização , Humanos , Indonésia , Lactente , Masculino , Norovirus/genética , Norovirus/isolamento & purificação , Filogenia , Rotavirus/genética , Rotavirus/isolamento & purificação , Análise de Sequência de RNA/métodos
12.
Int J Nanomedicine ; 14: 1865-1876, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30880985

RESUMO

Background: Rotavirus is the representative cause of severe acute gastroenteritis in young children. A characteristic feature of rotavirus is low infectious dose and robustness of the virion, suggesting sanitation and hygiene will have little impact. Thus, development of a vaccine should be given priority. Efficient capture of infectious viruses is an important step in generating a vaccine. Previously, antibody-integrated magnetic nanobeads (MNBs) have been developed for virus capture. This study examines the applicability of this method for infectious rotavirus recovery and enrichment. Materials and methods: Graphite-encapsulated MNBs were treated with radio frequency (RF) excited Ar/NH3 gas mixture plasma to introduce amino groups onto their surfaces. Rotavirus viral protein 7 (VP7) antibody was attached to the surface of MNBs via these amino groups using a coupling agent, N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP). The antibody-integrated MNBs were incubated with rotavirus-infected cell lysate and then separated from the supernatant by applying a magnetic field. After thorough washing, rotavirus was recovered and enrichment analysis done by polymerase chain reaction (PCR), immunochromatography, and an infection analysis using MA104 cells. Results and discussion: Immunochromatography and PCR indicate that anti-rotavirus antibody-integrated MNPs efficiently capture rotavirus with the capsid protein and viral RNA. The estimated recovery rate was 80.2% by PCR and 90.0% by infection analysis, while the concentrating factor was 7.9-fold by PCR and 6.7-fold by infection analysis. In addition, the absence of non-specific binding to the antibody-integrated MNPs was confirmed using anti-dengue virus antibody-integrated MNBs as a negative control. Conclusion: These results suggest that this capture procedure is a useful tool for recovery and enrichment of infectious rotavirus. Moreover, when combined with a suitable virus assay this capture procedure can increase the sensitivity of rotavirus detection. Therefore, this capture method is a valuable tool for generating vaccines as well as for developing sensitive detection systems for viruses.


Assuntos
Amônia/química , Anticorpos Antivirais/metabolismo , Argônio/química , Grafite/química , Magnetismo/métodos , Nanosferas/química , Gases em Plasma/química , Rotavirus/isolamento & purificação , Adsorção , Animais , Proteínas do Capsídeo/metabolismo , Linhagem Celular , Macaca mulatta , Reação em Cadeia da Polimerase , RNA Viral/metabolismo , Rotavirus/genética , Rotavirus/patogenicidade , Infecções por Rotavirus/virologia
13.
Jpn J Infect Dis ; 72(4): 256-260, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-30814461

RESUMO

The emergence of unusual DS-1-like intergenogroup reassortant rotaviruses with a bovine-like G8 genotype (DS-1-like G8P[8] strains) has been reported in several Asian countries. During the rotavirus surveillance program in Japan in 2017, a DS-1-like G8P[8] strain (RVA/Human-wt/JPN/SO1162/2017/G8P[8]) was identified in 43 rotavirus-positive stool samples. Strain SO1162 was shown to have a unique genotype constellation, including genes from both genogroup 1 and 2: G8-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2. Phylogenetic analysis revealed that the VP1 gene of strain SO1162 appeared to have originated from DS-1-like G1P[8] strains from Thailand and Vietnam, while the remaining 10 genes were closely related to those of previously reported DS-1-like G8P[8] strains. Thus, SO1162 was suggested to be a reassortant strain that acquired the VP1 gene from Southeast Asian DS-1-like G1P[8] strains on the genetic background of co-circulating DS-1-like G8P[8] strains. Our findings provide important insights into the evolutionary dynamics of emerging DS-1-like G8P[8] strains.


Assuntos
Vírus Reordenados/genética , Infecções por Rotavirus/virologia , Rotavirus/genética , Animais , Bovinos , Pré-Escolar , Evolução Molecular , Fezes/virologia , Genes Virais/genética , Genoma Viral/genética , Genótipo , Humanos , Japão , Filogenia , RNA Viral/genética , Vírus Reordenados/classificação , Vírus Reordenados/isolamento & purificação , Rotavirus/classificação , Rotavirus/isolamento & purificação , Análise de Sequência de DNA
14.
Arch Virol ; 164(4): 1229-1232, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30810805

RESUMO

Group A rotaviruses (RVAs) are important zoonotic pathogens that cause intestinal disease in humans and other mammals. In this study, the novel strain RVA/Pig/China/SC11/2017/G9P[23](SC11) was isolated from fecal samples from a pig farm in Sichuan province, southwestern China. The complete genome was found to be 18,347 bp in length with 11 segments. The genotype constellation of strain SC11 was G9-P[23]-I12-R1-C1-M1-A1-N1-T1-E1-H1, according to whole-genome sequencing analysis. The VP1, VP2, VP4, VP6, NSP1-NSP3, and NSP5 genes of RVA strain SC11 were found to be closely related to those of porcine and/or porcine-like human RVAs. Meanwhile, the VP7 and NSP4 genes of strain SC11 were closely related to genes of human RVAs. However, it was difficult to pinpoint the porcine or human origin of the VP3 gene of strain SC11 based on the available data. These results showed that SC11 originated from a natural reassortment event between human and pig RVA strains, and crossover points for recombination were identified at nucleotides (nt) 109-806 of NSP2. This is the first report of such a reassortant and recombinant RVA strain in the southwestern region of China.


Assuntos
Vírus Reordenados/isolamento & purificação , Recombinação Genética , Infecções por Rotavirus/veterinária , Infecções por Rotavirus/virologia , Rotavirus/genética , Doenças dos Suínos/virologia , Animais , Genoma Viral , Genótipo , Humanos , Filogenia , Vírus Reordenados/classificação , Vírus Reordenados/genética , Rotavirus/classificação , Rotavirus/isolamento & purificação , Suínos
15.
J Vet Intern Med ; 33(2): 918-922, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30788861

RESUMO

BACKGROUND: Currently, diagnosis of equine coronavirus (ECoV) relies on the exclusion of other infectious causes of enteric disease along with molecular detection of ECoV in feces or tissue. Although this approach is complete, it is costly and may not always be achievable. OBJECTIVE: We hypothesized that the overall fecal shedding of ECoV in hospitalized horses is low. Our objective was to determine whether systemically healthy horses and horses with gastrointestinal disorders shed ECoV in their feces at the time of admission to a referral hospital and after 48 hours of stress associated with hospitalization. ANIMALS: One-hundred thirty adult horses admitted to the Washington State University Veterinary Teaching Hospital for gastrointestinal disease (n = 65) or for imaging under anesthesia (n = 65) that were hospitalized for 48 hours. Owner consent was obtained before sampling. METHODS: Fecal samples were collected at admission and 48 hours later. Polymerase chain reaction (PCR) for ECoV and electron microscopy (EM) were performed on all samples. RESULTS: Only 1 of 258 fecal samples was PCR-positive for ECoV. Electron microscopy identified ECoV-like particles in 9 of 258 samples, parvovirus-like particles in 4 of 258 samples, and rotavirus-like particles in 1 of 258 samples. CONCLUSIONS AND CLINICAL IMPORTANCE: The presence of ECoV in feces of hospitalized adult horses was low. Thus, fecal samples that are PCR-positive for ECoV in adult horses that have clinical signs consistent with this viral infection are likely to be of diagnostic relevance. The clinical relevance of the viruses observed using EM remains to be investigated.


Assuntos
Betacoronavirus 1/isolamento & purificação , Fezes/microbiologia , Doenças dos Cavalos/microbiologia , Animais , Fezes/virologia , Gastroenteropatias/veterinária , Doenças dos Cavalos/virologia , Cavalos , Hospitalização , Microscopia Eletrônica , Parvovirus/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Rotavirus/isolamento & purificação , Washington
16.
Food Environ Virol ; 11(1): 65-75, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30607905

RESUMO

Global burden of acute viral gastroenteritis remains high, particularly in developing countries including Bangladesh. Sewage water (SW) is an important node to monitor enteric pathogens both in the environment and among the population. Analysis of SW in Dhaka city deems crucially important because a large number of urban-city dwellers live in Dhaka city, the capital of Bangladesh, under a constant threat of precarious sewerage system. In this study, we collected raw SW from five locations of Dhaka city every month from June 2016 to May 2017. It was concentrated with polyethylene glycol (PEG) and investigated for three major enteric viruses, rotavirus A (RVA), norovirus GII (NoV GII) and adenovirus (AdV) using polymerase chain reaction (PCR). Most of these SW samples collected from both hospitals and non-hospital areas yielded enteric viruses: 76% samples were positive for AdV, followed by 53% NoV GII and 38% RVA. Viral load was determined as much as 1 × 107 copies/ml for RVA and 3.5 × 103 copies/ml for NoV GII. Importantly, NoV GII and AdV that can affect people of all ages were predominated during monsoon also when SW overflows and spreads over a wide and crowded area. Genotypes G1, G2, G3, G8, and G9 for RVA, GII.4 for NoV, and type 41 for AdV were detected representing the current profile of circulating genotypes in the population. This study provides the first evidence of distribution of major diarrheal viruses in SW in Dhaka city which is alarming showing grave risk of impending outbreaks through exposure.


Assuntos
Adenoviridae/genética , Norovirus/genética , Rotavirus/genética , Esgotos/virologia , Adenoviridae/classificação , Adenoviridae/isolamento & purificação , Bangladesh , Humanos , Resíduos de Serviços de Saúde , Epidemiologia Molecular/métodos , Tipagem Molecular , Norovirus/classificação , Norovirus/isolamento & purificação , Filogenia , Rotavirus/classificação , Rotavirus/isolamento & purificação
17.
Int J Infect Dis ; 80: 66-72, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30639406

RESUMO

OBJECTIVES: Enteric viruses are responsible for foodborne and waterborne infections affecting a large number of people. Data on food and water viral contamination in the south of Italy (Sicily) are scarce and fragmentary. The aim of this study was to evaluate the presence of viral contamination in food, water samples, and surface swabs collected in Sicily METHODS: The survey was conducted on 108 shellfish, 23 water samples (seawater, pipe water, and torrent water), 52 vegetables, one peach and 17 berries, 11 gastronomic preparations containing fish products and/or raw vegetables, and 28 surface swabs. Hepatitis A virus (HAV), genogroup GI, GII, and GIV norovirus (NoV), enterovirus (EV), rotavirus (RoV), hepatitis E virus (HEV), adenovirus (AdV), and bocavirus (BoV) were detected by nested (RT) PCR, real-time PCR, and sequence analysis. RESULTS: The most frequently detected viruses in shellfish were HAV (13%), NoV (18.5%), and EV (7.4%). Bocavirus was found in 3.7%, HEV in 0.9%, and AdV in 1.9% of the molluscs. Of the 23 water samples, 21.7% were positive for GII NoV and 4.3% for RoV and HEV genotype 3. Of the 70 vegetable samples, 2.9% were positive for NoV GI (GI.5 and GI.6), 2.9% for EV, and 1.4% for HEV. In the gastronomic preparations, only one EV (9%) was detected. No enteric viruses were detected in the berries, fruit, or swabs analyzed. CONCLUSIONS: Molecular surveillance of water and food samples clearly demonstrated that human pathogenic viruses are widely found in aquatic environments and on vegetables, and confirmed the role of vegetables and bivalve molluscs as the main reservoirs.


Assuntos
Enterovirus/isolamento & purificação , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Microbiologia da Água , Animais , DNA Viral/isolamento & purificação , Água Potável/virologia , Frutas/virologia , Vírus da Hepatite A/isolamento & purificação , Vírus da Hepatite E/isolamento & purificação , Humanos , Norovirus/isolamento & purificação , Rotavirus/isolamento & purificação , Alimentos Marinhos/virologia , Frutos do Mar/virologia , Sicília , Verduras/virologia , Poluição da Água
18.
Food Environ Virol ; 11(1): 9-19, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30560490

RESUMO

We investigated the present forms of genogroup II norovirus and group A rotavirus in surface water used for drinking water production. River water samples (N = 15) collected at a drinking water treatment plant (DWTP) monthly from June 2017 to August 2018 were fractioned by filtration through 10- and 0.45-µm-pore-size membranes, and viruses present in suspended and dissolved forms were quantitatively detected. Norovirus GII was present in > 10-µm- and 0.45-10-µm-suspended and dissolved forms with detection rates of 33%, 60%, and 87%, respectively. Rotavirus A was detected more frequently than norovirus GII in each form (> 10 µm suspended, 73%; 0.45-10 µm suspended, 93%; dissolved, 100%). We also analyzed surface water samples from 21 DWTPs all over Japan in non-epidemic and epidemic seasons of gastroenteritis. Norovirus GII was detected in 48% and 81% of samples with the concentrations of up to 4.1 and 5.3 log10 copies/L in dissolved form in non-epidemic and epidemic seasons, respectively, and GII.4 Sydney 2012 was predominant genotype followed by GII.2. Rotavirus A was detected in 95% and 86% of samples with the maximum concentrations of 5.5 and 6.3 log10 copies/L in dissolved form in respective seasons. Concentration of norovirus GII was similar in 0.45-10-µm suspended and dissolved forms, while there was a significant difference for rotavirus A (P < 0.01, pared t test), indicating that rotavirus A was less associated with suspended solids in the surface water samples compared to norovirus GII. Our observations provide important implications for understanding of viral behavior in environmental waters.


Assuntos
Água Potável/virologia , Norovirus , Rotavirus , Filtração , Japão , Norovirus/genética , Norovirus/isolamento & purificação , Reação em Cadeia da Polimerase , RNA Viral/genética , RNA Viral/isolamento & purificação , Rios/virologia , Rotavirus/genética , Rotavirus/isolamento & purificação , Purificação da Água
19.
Ann Lab Med ; 39(1): 50-57, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30215230

RESUMO

BACKGROUND: The Automated Fluorescent Immunoassay System (AFIAS) rotavirus assay (Boditech Med Inc., Chuncheon, Korea) is a new rapid antigen test for rotavirus detection. We evaluated the performance of this assay for detecting rotaviruses and their specific genotypes in clinical stool samples. METHODS: AFIAS rotavirus assay was performed in 103 rotavirus-positive and 103 rotavirus-negative stool samples (confirmed by both PCR and ELISA), and its results were compared with those of PCR, ELISA, and immunochromatographic assay (ICA). We evaluated diagnostic sensitivity/specificity, the detectability of rotavirus subtypes, lower limit of detection (LLOD), reproducibility, cross-reactivity, and interference of AFIAS rotavirus assay. RESULTS: Based on PCR and ELISA results, diagnostic sensitivity and specificity of the AFIAS rotavirus assay were both 99.0%. LLOD results showed that the AFIAS assay had sensitivity similar to or greater than ICA and ELISA. High reproducibility was confirmed, and no cross-reactivity or interference was detected. This assay could detect genotypes G1P[8], G2P[4], G3P[8], G4P[6], G4P[8], G8P[4], G8P[8], G9P[4], and G9P[8]. CONCLUSIONS: The AFIAS rotavirus assay showed high reproducibility, sensitivity, and specificity as well as excellent agreement with ELISA, PCR, and ICA. It detected the most common as well as unusual genotypes of rotavirus prevalent in Korea. It could be a useful on-site assay for rapid, convenient, and cost-effective detection of rotavirus infection.


Assuntos
Antígenos Virais/análise , Imunofluorescência/métodos , Rotavirus/metabolismo , Antígenos Virais/imunologia , Automação , Criança , Pré-Escolar , Fezes/virologia , Imunofluorescência/instrumentação , Genótipo , Humanos , Lactente , Recém-Nascido , Limite de Detecção , Kit de Reagentes para Diagnóstico , Reprodutibilidade dos Testes , Rotavirus/genética , Rotavirus/isolamento & purificação , Infecções por Rotavirus/diagnóstico , Infecções por Rotavirus/virologia
20.
Sci Total Environ ; 651(Pt 1): 298-308, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30240914

RESUMO

In this study, total coliforms (TC), Escherichia coli, enterovirus (EV), rotavirus (RV), and human mastadenovirus species C and F (HAdV-C and HAdV-F) were evaluated in water samples from Belo Stream. For HAdV-C and F, the infectivity was assessed by integrated cell culture quantitative real-time polymerase chain reaction (ICC-qPCR). Samples were collected monthly (May/2015 to April/2016) at four sites. Viral analyses were performed for both ultracentrifuge-concentrated and unconcentrated samples. For site P4 (used for recreational purposes), QMRA was applied to estimate health risks associated with exposure to E. coli and HAdV-C and F. TC and E. coli were present throughout the collection period. EV and RV were not detected. HAdV-C were present in 8.51% (1.89E + 06 to 2.28E + 07 GC (Genomic Copies)/L) and 21.27% (2.36E + 05 to 1.29E + 07 GC/L) for unconcentrated and concentrated samples, respectively. For HAdV-F were 12.76% (2.77E + 07 to 3.31E + 08 GC/L) and 48.93% (1.10E + 05 to 4.50E + 08 GC/L) for unconcentrated and concentrated samples, respectively. For unconcentrated samples, infectivity for HAdV-C was detected in 37.20% (1st ICC-qPCR) and 25.58% (2nd ICC-qPCR). For HAdV-F, infectivity was detected in 6.97% (1st ICC-qPCR) and 6.97% (2nd ICC-qPCR). For concentrated samples, HAdV-C infectious was observed in 17.02% (1st ICC-qPCR) and in 8.51% (2nd ICC-qPCR). For HAdV-F, were present in 8.51% for both 1st and 2nd ICC-qPCR. Statistical analyzes showed significant difference between the collection sites when analyzed the molecular data of HAdV-F, data of TC and E. coli. Correlation tests showed direct correlation between HAdV-F with E. coli and TC. E. coli concentrations translated to the lowest estimates of infection risks (8.58E-05 to 2.17E-03). HAdV-F concentrations were associated with the highest infection risks at 9.99E-01 and for group C, 1.29E-01 to 9.99E-01. These results show that commonly used bacterial indicators for water quality may not infer health risks associated with viruses in recreational freshwaters.


Assuntos
Medição de Risco , Rios/microbiologia , Qualidade da Água , Adenovírus Humanos/isolamento & purificação , Brasil , Enterobacteriaceae/isolamento & purificação , Enterovirus/isolamento & purificação , Escherichia coli/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Recreação , Rios/virologia , Rotavirus/isolamento & purificação
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