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1.
Viruses ; 13(2)2021 01 26.
Artigo em Inglês | MEDLINE | ID: mdl-33530573

RESUMO

Sapovirus is a common cause of acute gastroenteritis in all age groups. Sapovirus infections are seldom investigated in Spain, and its epidemiology in the country is not well known. The use of molecular diagnostic procedures has allowed a more frequent detection of sapoviruses in patients with diarrhea. A total of 2545 stool samples from patients with acute gastroenteritis attended from June 2018 to February 2020 at the Clinic University Hospital in Valencia, Spain, were analyzed by reverse transcription (RT) and real-time multiplex PCR (RT-PCR) to investigate the etiology of enteric infections. Sapovirus was the second enteric virus detected with a positive rate of 8%, behind norovirus (12.2%) and ahead of rotavirus (7.1%), astrovirus (4.9%) and enteric adenoviruses (2.9%). Most sapovirus infections occurred in infants and young children under 3 years of age (74%) with the highest prevalence in autumn and early winter. Coinfections were found in 25% of the patients with sapovirus diarrhea, mainly with other enteric viruses. Genotyping demonstrated the circulation of seven different genotypes during the study period, with a predominance of genotypes GI.1, GI.2, and GII.1. Phylogenetic analysis showed that genogroup GII strains form a cluster separated from genogroup GI and GV, being genotype GV.1 strains related to genotype GI.1 and GI.2 strains.


Assuntos
Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Gastroenterite/epidemiologia , Gastroenterite/virologia , Sapovirus/genética , Fatores Etários , Infecções por Caliciviridae/diagnóstico , Coinfecção/epidemiologia , Coinfecção/microbiologia , Coinfecção/virologia , Diarreia/diagnóstico , Diarreia/epidemiologia , Diarreia/virologia , Feminino , Gastroenterite/diagnóstico , Variação Genética , Genótipo , Humanos , Masculino , Epidemiologia Molecular , Reação em Cadeia da Polimerase Multiplex , Filogenia , Prevalência , RNA Viral/genética , Sapovirus/classificação , Sapovirus/isolamento & purificação , Estações do Ano , Espanha/epidemiologia
2.
Talanta ; 225: 121978, 2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33592726

RESUMO

In modern times, viruses still threaten people's lives. Among them, norovirus was the main pathogenic factor in the cause of gastroenteritis and foodborne illness, of which the GII.4 and GII.17 genotypes are prevalent in China and most parts of the world. A simple and low-cost platform for rapid and accurate norovirus detection remains a major challenge. After the cell-free system and paper-based chromogenic system were optimized, a rapid and specific norovirus detection method was established based on norovirus-specific sequences in combination with toehold switch elements. The development of a visible color change during detection eliminates the need for any complicated instruments. We validated this strategy and its specificity in differentiating GII.4, GII.17, Zika virus, and human coronavirus HKU1. The results showed that the optimized detection system not only provided a simple and rapid detection method for the sufficient differentiation of the two norovirus genotypes but also showed high specificity and no cross-reactivity with other viruses. Using nucleic acid isothermal amplification, this assay showed a limit of detection of 0.5 pM for the GII.4 genotype and 2.6 fM for the GII.17 genotype in reactions that could be observed directly with the naked eye. Our results suggested that this paper-based colorimetric method could serve as a simple and low-cost visual detection method for pathogens in clinical samples, especially in remote or rural areas.


Assuntos
Infecções por Caliciviridae/diagnóstico , Colorimetria/métodos , Gastroenterite/diagnóstico , Infecções por Caliciviridae/virologia , Colorimetria/economia , Colorimetria/instrumentação , Análise Custo-Benefício , Gastroenterite/virologia , Genótipo , Humanos , Norovirus/genética , Norovirus/fisiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Papel , RNA Viral/genética , Sensibilidade e Especificidade
3.
BMJ Case Rep ; 14(1)2021 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-33462001

RESUMO

A 75-year-old man was admitted with a 3-month history of worsening diarrhoea and weight loss. He was on long-term immunosuppression following cardiac transplantation. Investigations revealed herpes simplex oesophagitis and stool samples were positive for norovirus. Treatment with acyclovir and nitazoxanide resulted in a complete resolution of symptoms. Norovirus is a common cause of infectious gastroenteritis, but immunosuppressed patients may present with chronic diarrhoea rather than an acute illness. This case highlights the importance of a low clinical threshold for testing for norovirus infection in immunocompromised patients.


Assuntos
Infecções por Caliciviridae/imunologia , Diarreia/virologia , Gastroenterite/imunologia , Hospedeiro Imunocomprometido , Norovirus/isolamento & purificação , Complicações Pós-Operatórias/imunologia , Perda de Peso , Idoso , Infecções por Caliciviridae/complicações , Infecções por Caliciviridae/diagnóstico , Doença Crônica , Diarreia/imunologia , Gastroenterite/complicações , Gastroenterite/diagnóstico , Transplante de Coração , Humanos , Masculino , Complicações Pós-Operatórias/diagnóstico , Perda de Peso/imunologia
4.
BMC Infect Dis ; 21(1): 54, 2021 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-33435906

RESUMO

BACKGROUND: An outbreak of acute gastroenteritis occurred in a kindergarten located Shenzhen City on March 4, 2018. We were invited to investigate to the risk factors associated with this outbreak. METHODS: We conducted retrospective cohort-studies on three different groups of subjects in order to figure out the difference of incidence of acute gastroenteritis among subjects of different activities on March 2: group one consisted of people who attended the Lantern festival activities; group two consisted of children and employees who ate breakfast and bread provided by the kindergarten; and groups three consisted of children and employees who did not eat breakfast or bread provided by the kindergarten. Fecal, anal swabs, dishware swabs and hand swabs specimens were collected in the study. Bacteria known to cause acute gastroenteritis were cultured. Viruses associated with acute gastroenteritis were tested using real-time PCR. Capsid gene fragment of 557 bp of norovirus was amplified and sequenced. The phylogenetic tree was constructed with MEGA 7.0 using neighbor-joining method based on capsid gene fragment of norovirus. RESULTS: A total of 143 suspected cases were identified in this outbreak. Diarrhea happened more often in adults than in children while emesis and bellyache were more frequently found in children than in adults. Higher AGE incidence was observed in group 2, children and employees who had breakfast in the kindergarten on March 2, as well as in group 3, and among employees who eating bread involved in breakfast provided on March 2. Five anal swab specimens were positive for norovirus. All noroviruses belongs to group II.3 and have an identity more than 99%. CONCLUSION: A chef, as an asymptomatic carrier with norovirus, was the infectious resource in this outbreak. He contaminated breakfast food provided on March 2. Although morning check is implemented in kindergartens of China, employees are often excluded in morning check. Our finding highlights the importance of morning check covering employees and periodical training for cooks.


Assuntos
Desjejum , Infecções por Caliciviridae/epidemiologia , Portador Sadio/virologia , Surtos de Doenças , Manipulação de Alimentos , Gastroenterite/epidemiologia , Norovirus/genética , Escolas Maternais , Adulto , Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/prevenção & controle , Infecções por Caliciviridae/virologia , Criança , Pré-Escolar , China/epidemiologia , Diarreia/epidemiologia , Diarreia/virologia , Fezes/virologia , Feminino , Microbiologia de Alimentos/métodos , Gastroenterite/diagnóstico , Gastroenterite/prevenção & controle , Gastroenterite/virologia , Humanos , Incidência , Masculino , Filogenia , Quarentena/métodos , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Fatores de Risco , Vômito/epidemiologia , Vômito/virologia
5.
Arch Virol ; 165(12): 2767-2776, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32949263

RESUMO

Human norovirus is the leading cause of viral gastroenteritis worldwide. Rapid detection facilitates management of disease outbreaks, but field diagnosis is difficult to achieve due to the lack of reliable and portable methods. Recombinase polymerase amplification (RPA) is a robust isothermal amplification method that is capable of rapidly amplifying and detecting nucleic acids using simple equipment. In this study, RPA combined with lateral flow (LF) strips specific for human genogroup II (GII) noroviruses was established and evaluated. The assay specifically detects purified GII noroviruses as well as RNA in boiled human stool samples, with a sensitivity of 50 norovirus genome copies per reaction. The whole detection procedure of the one-step RT-RPA-LF is completed within 20 min, which is eight times faster than that of the standard real-time RT-PCR. The RT-RPA-LF method described here is suitable for rapid field diagnosis of all GII noroviruses in human stool samples.


Assuntos
Infecções por Caliciviridae/diagnóstico , Norovirus/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Infecções por Caliciviridae/genética , Fezes/virologia , Humanos , Norovirus/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Recombinases/química , Sensibilidade e Especificidade
6.
Arch Virol ; 165(10): 2335-2340, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32719956

RESUMO

Sapoviruses are increasingly being recognized as pathogens associated with gastroenteritis in humans. Human sapoviruses are currently assigned to 18 genotypes (GI.1-7, GII.1-8, GIV.1, and GV.1-2) based on the sequence of the region encoding the major structural protein. In this study, we evaluated 11 polymerase chain reaction (PCR) assays using published and newly designed/modified primers and showed that four PCR assays with different primer combinations amplified all of the tested human sapovirus genotypes using either synthetic DNA or cDNA prepared from human sapovirus-positive fecal specimens. These assays can be used as improved broadly reactive screening tests or as tools for molecular characterization of human sapoviruses.


Assuntos
Infecções por Caliciviridae/virologia , Primers do DNA/química , Gastroenterite/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sapovirus/genética , Proteínas Estruturais Virais/genética , Sequência de Bases , Infecções por Caliciviridae/diagnóstico , Primers do DNA/genética , Fezes/virologia , Gastroenterite/diagnóstico , Expressão Gênica , Genótipo , Humanos , Tipagem Molecular/métodos , Filogenia , Sapovirus/classificação , Sapovirus/isolamento & purificação , Alinhamento de Sequência
7.
Int J Food Microbiol ; 333: 108787, 2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-32702583

RESUMO

Human noroviruses (HuNoVs) are the primary non-bacterial pathogens causing acute gastroenteritis worldwide. Here we reported a co-infection of HuNoVs with different genotypes during an outbreak of gastroenteritis in travelers. The aim was to trace the source and transmission patterns of the infections using next-generation sequencing (NGS). An investigation was conducted on a cross-border travel group who came back to China from Thailand for symptoms of gastroenteritis. Anal swabs were collected from 23 people and samples were analyzed using RT-qPCR. A total of 11 samples tested positive for HuNoVs. All samples tested negative for bacterial pathogens in the surveillance list. Positive samples for HuNoVs were further analyzed using NGS. Seven out of 11 positive samples were sequenced and 16 viral genome sequences for 10 different strains of HuNoVs were obtained. We demonstrated that the outbreak was associated with co-infection of multiple genotypes of HuNoVs and the source of infections was probably contaminated water or food. Besides, four different HuNoVs genotypes (GI.5[P12], GIX.1[GII·P15], GI.7[P7] and GII.8[P8]) were identified in one patient. Co-infection with both genogroup GI and GII, and co-infection with two different P types ([P10] and [P13]) of genotype GI.3 were identified in different patients. Findings from this study show that individuals can be simultaneously infected with multiple strains of HuNoVs and NGS can help investigating these issues. Further, this study shows that food and water are potential vehicles for transmission of multiple foodborne viruses.


Assuntos
Infecções por Caliciviridae/diagnóstico , Doenças Transmitidas por Alimentos/virologia , Gastroenterite/diagnóstico , Norovirus/genética , Norovirus/isolamento & purificação , Adolescente , Adulto , Sequência de Bases , Infecções por Caliciviridae/virologia , Criança , China/epidemiologia , Surtos de Doenças , Feminino , Gastroenterite/virologia , Genoma Viral/genética , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Metagenômica , Pessoa de Meia-Idade , Filogenia , Reação em Cadeia da Polimerase em Tempo Real , Tailândia , Viagem , Adulto Jovem
8.
Zhonghua Liu Xing Bing Xue Za Zhi ; 41(4): 547-551, 2020 Apr 10.
Artigo em Chinês | MEDLINE | ID: mdl-32344480

RESUMO

Objective: To analyze the epidemiological characteristics related to Norovirus-associated infectious diarrhea bud events in Shanghai, and to discuss the value of bud events surveillance, so as to help Norovirus-associated infectious diarrhea control, prevention and to improve the surveillance system on bud events. Methods: Data related to 142 Norovirus-associated infectious diarrhea bud events were gathered from 16 districts of Shanghai areas in 2018 and were analyzed. Results: There were 2 peaks, April and November for 142 Norovirus-associated infectious diarrhea bud events reported in 2018. 98.59% (140/142) of the bud events occurred in schools and kindergartens. 80.28% (114/142) of the bud events would involve 6-19 cases per each event. The median time duration between the onset date of the first case and the reporting date of the event was 2 days and the median event duration was 7 days. The bud event scale and regional incidence of bud events as well as the reporting timelines were influential factors on the duration of the bud events. Conclusions: Two peaks, spring and winter bud events of the Norovirus-associated infectious diarrhea were seen in Shanghai in 2018. Schools and kindergartens were to be prioritized locations for control and prevention of Norovirus-associated infectious diarrhea bud events. Surveillance system was proved to have contributed to the early detection, reporting and control of bud events.


Assuntos
Infecções por Caliciviridae/epidemiologia , Diarreia/epidemiologia , Diarreia/virologia , Norovirus/isolamento & purificação , Infecções por Caliciviridae/diagnóstico , China/epidemiologia , Diarreia/diagnóstico , Disenteria/epidemiologia , Humanos
9.
Epidemiol Infect ; 148: e52, 2020 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-32063241

RESUMO

Norovirus (NoV) infections occur very frequently yet are rarely diagnosed. In Denmark, NoV infections are not under surveillance. We aimed to collect and describe existing laboratory-based NoV data. National NoV laboratory data were collected for 2011-2018, including information on patient identification number, age and sex, requesting physician, analysis date and result. We defined positive patient-episodes by using a 30-day time window and performed descriptive and time series analysis. Diagnostic methods used were assessed through a survey. We identified 15 809 patient-episodes (11%) out of 142 648 tested patients with an increasing trend, 9366 in 2011 vs. 32 260 in 2018. This corresponded with a gradual introduction of polymerase chain reaction analysis in laboratories. The highest positivity rate was in patients aged <5 years (15%) or >85 years (17%). There was a large difference in test performance over five Danish geographical regions and a marked seasonal variation with peaks from December to February. This is the first analysis of national NoV laboratory data in Denmark. A future laboratory-based surveillance system may benefit public health measures by describing trend, burden and severity of seasons and possibly pinpoint hospital outbreaks.


Assuntos
Infecções por Caliciviridae , Surtos de Doenças , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/epidemiologia , Criança , Pré-Escolar , Dinamarca/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Estações do Ano , Adulto Jovem
10.
Eur J Clin Microbiol Infect Dis ; 39(1): 103-112, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31506730

RESUMO

The Hologic Panther Fusion® Open Access™ functionality allows implementation of laboratory-developed tests (LDTs), with fully automated sample extraction, real-time PCR, and result interpretation. We report the development and validation of a multiplex LDT for norovirus G1, norovirus G2, and rotavirus from stool samples on this system. The LDT was optimized for primer and probe sequences, salt concentration, and PCR annealing temperature. Reproducibility of the PCR and extraction process was assessed. Performance of the multiplex LDT assay was evaluated with external quality assessment (EQA) samples and compared to a commercial multiplex assay (Allplex™ GI-Virus Assay, Seegene) in clinical samples. Salt concentrations and annealing/extension temperature were optimized to 4 mM MgCl2, 70 mM KCl, 20 mM Tris, and 60 °C, respectively. The user-prepared part of the LDT PCR mix (containing salts, probes, and primers) was stable for ≥ 11 days onboard the instrument. We observed reproducible results of PCR and the extraction process. The LDT had a sensitivity comparable to or greater than the commercial Allplex™ assay and showed excellent linearity. Forty-five EQA samples yielded the expected result with the LDT. There was 100% concordance between LDT and Allplex™ results in 160 clinical samples. Results from the suspension and direct swab stool sample preparation methods were highly concordant in the LDT. We report the successful development and validation of a multiplex PCR LDT for detection of norovirus G1, norovirus G2, and rotavirus from stool samples on the Panther Fusion® system.


Assuntos
Infecções por Caliciviridae/diagnóstico , Gastroenterite/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Norovirus/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecções por Rotavirus/diagnóstico , Rotavirus/isolamento & purificação , Automação Laboratorial/normas , Primers do DNA , Fezes/virologia , Gastroenterite/virologia , Genótipo , Humanos , RNA Viral/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
11.
Transbound Emerg Dis ; 67(2): 822-833, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31665828

RESUMO

The lagovirus rabbit haemorrhagic disease virus (RHDV) has been circulating in Australia since the mid-1990s when it was released to control overabundant rabbit populations. In recent years, the viral diversity of different RHDVs in Australia has increased, and currently four different types of RHDV are known to be circulating. To allow for ongoing epidemiological studies and impact assessments of these viruses on Australian wild rabbit populations, it is essential that serological tools are updated. To this end, reference sera were produced against all four virulent RHDVs (RHDV, RHDV2 and two different strains of RHDVa) known to be present in Australia and tested in a series of available immunological assays originally developed for the prototype RHDV, to assess patterns of cross-reactivity and the usefulness of these assays to detect lagovirus antibodies, either in a generic or specific manner. Enzyme-linked immunosorbent assays (ELISAs) developed to detect antibody isotypes IgM, IgA and IgG were sufficiently cross-reactive to detect antibodies raised against all four virulent lagoviruses. For the more specific detection of antibodies to the antigenically more different RHDV2, a competition ELISA was adapted using RHDV2-specific monoclonal antibodies in combination with Australian viral antigen. Archival serum banks from a long-term rabbit monitoring site where rabbits were sampled quarterly over a period of 6 years were re-screened using this assay and revealed serological evidence for the arrival of RHDV2 in this population at least 5 months prior to its initial detection in Australia in a dead rabbit in May 2015. The serological methods and reference reagents described here will provide valuable tools to study presence, prevalence and impact of RHDV2 on Australian rabbit populations; however, the discrimination of different antigenic variants of RHDVs as well as mixed infections at the serological level remains challenging.


Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Infecções por Caliciviridae/veterinária , Vírus da Doença Hemorrágica de Coelhos/imunologia , Animais , Austrália/epidemiologia , Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática/veterinária , Vírus da Doença Hemorrágica de Coelhos/isolamento & purificação , Coelhos , Estudos Retrospectivos
12.
Acta Med Acad ; 49(3): 278-280, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33781071

RESUMO

Norovirus is a substantial burden on the U.S. We compared norovirus outbreaks before and during COVID-19. There were fewer norovirus outbreaks during COVID-19 compared to a similar time period in 2019 (326 versus 638, P<0.001). CONCLUSION: COVID-19 public health interventions may be considered to decrease the burden of norovirus. This demonstrates the ability of more restrictive interventions to decrease other outbreaks of known or emerging viruses.


Assuntos
Infecções por Caliciviridae , Controle de Doenças Transmissíveis , Norovirus/isolamento & purificação , /epidemiologia , Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/prevenção & controle , Controle de Doenças Transmissíveis/métodos , Controle de Doenças Transmissíveis/organização & administração , Controle de Doenças Transmissíveis/estatística & dados numéricos , Transmissão de Doença Infecciosa/prevenção & controle , Humanos , Saúde Pública/métodos , Saúde Pública/estatística & dados numéricos , Estados Unidos/epidemiologia
13.
Sci Rep ; 9(1): 19576, 2019 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-31862970

RESUMO

Norovirus is one of the most common causes of gastroenteritis. Following infection, anti-norovirus salivary immunoglobulin G (IgG) rises steeply within 2 weeks and remains elevated for several months; this immunoconversion can serve as an indicator of infection. We used a multiplex salivary immunoassay to study norovirus infections among 483 visitors to a Lake Michigan beach in 2015. Saliva was collected on the day of the beach visit (S1); after 10-14 days (S2); and after 30-40 days (S3). Luminex microspheres were coupled to recombinant antigens of genogroup I (GI) and II (GII) noroviruses and incubated with saliva. Immunoconversion was defined as at least 4-fold increase in anti-norovirus IgG antibody response from S1 to S2 and a 3-fold increase from S1 to S3. Ten (2.1%) immunoconverted to either GI (2) or GII (8) norovirus. Among those who immunoconverted, 40% reported at least one gastrointestinal symptom and 33% reported diarrhea, compared to 15% (p = 0.06) and 8% (p = 0.04) among those who did not immunoconvert, respectively. The two participants who immunoconverted to GI norovirus both swallowed water during swimming (p = 0.08). This study demonstrated the utility of a non-invasive salivary immunoassay to detect norovirus infections and an efficient approach to study infectious agents in large cohorts.


Assuntos
Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/virologia , Imunoensaio/métodos , Norovirus/patogenicidade , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Estudos Prospectivos , Adulto Jovem
15.
J Vet Diagn Invest ; 31(6): 909-912, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31650901

RESUMO

Swine diarrhea can be caused by multiple agents, including porcine epidemic diarrhea virus (PEDV), porcine sapelovirus (PSV), and porcine sapovirus (SaV). We designed a one-step triplex reverse-transcription PCR (RT-PCR) detection method including 3 pairs of primers that focused on the S1 gene of PEDV, a conserved gene of PSV, and the VP1 gene of SaV. The optimal concentrations of upstream and downstream primers in the triplex RT-PCR were 0.24 µM for PEDV, 0.15 µM for PSV, and 0.2 µM for SaV, and the optimal annealing temperature was 55.5°C. Triplex RT-PCR assessment of 402 piglet diarrhea samples was compared with conventional individual RT-PCR. Concordance rates in both tests for individual viruses were 100%, 97.6%, and 94.4% for PEDV, PSV, and SaV, respectively. PEDV, PSV, and SaV were detected in 57.2%, 10.4%, and 9.0% of the samples, respectively. The high sensitivity and specificity of this triplex RT-PCR-based detection method for PEDV, PSV, and SaV could allow rapid detection and analysis of mixed infections by these 3 viruses.


Assuntos
Infecções por Caliciviridae/veterinária , Infecções por Coronavirus/veterinária , Diarreia/veterinária , Reação em Cadeia da Polimerase Multiplex/veterinária , Infecções por Picornaviridae/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Doenças dos Suínos/diagnóstico , Animais , Infecções por Caliciviridae/diagnóstico , Infecções por Coronavirus/diagnóstico , Diarreia/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Picornaviridae/isolamento & purificação , Infecções por Picornaviridae/diagnóstico , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sapovirus/isolamento & purificação , Suínos , Doenças dos Suínos/virologia
16.
J Clin Virol ; 120: 51-56, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31563652

RESUMO

BACKGROUND: We undertook a prospective clinical study to evaluate PCR.Ai's (www.pcr.ai) accuracy and impact when automating the manual data-analysis and quality control steps associated with routine clinical pathogen testing using real-time PCR (qPCR). OBJECTIVES: We evaluated the impact of PCR.Ai when used as the final interpretation/verification step for routine in-house qPCR tests for respiratory pathogens and for norovirus for a total of 22,200 interpretations. STUDY DESIGN: We compared PCR.Ai to our existing manual interpretation, to determine accuracy and hands-on time savings. PCR.Ai was accurate. RESULTS AND CONCLUSIONS: There was 100% concurrence between validated respiratory virus and norovirus detection by our manual routine analysis method and PCR.Ai. Furthermore, there were significant routine savings with PCR.Ai of 45 min/respiratory run and 32 min/norovirus run. Our conclusion is that PCR.Ai is a highly accurate time-saving tool that reduces complexity of qPCR analysis and hence the need for specialists and hands-on time. It demonstrated capabilities to enable us to get results out more quickly with lower costs and less risk of errors.


Assuntos
Infecções por Caliciviridae/diagnóstico , Norovirus/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Infecções Respiratórias/virologia , Automação , Humanos , Técnicas de Diagnóstico Molecular/normas , Norovirus/isolamento & purificação , Estudos Prospectivos , RNA Viral/análise , Sensibilidade e Especificidade
17.
Zhonghua Liu Xing Bing Xue Za Zhi ; 40(8): 883-888, 2019 Aug 10.
Artigo em Chinês | MEDLINE | ID: mdl-31484248

RESUMO

Objective: To investigate the epidemiologic characteristics of Norovirus among adult patients suffering from infectious diarrhea in Shanghai, so as to provide evidence for developing related strategies on prevention and control of the disease. Methods: Diarrheal outpatients were monitored at the 'Intestinal clinic' from 22 hospitals involved in the sentinel surveillance program in Shanghai. Information on demographic and epidemiologic features of the patients was collected while data and clinical, fecal specimens were collected and sent to the district CDC for Norovirus detection. Positive rates of Norovirus were also compared in various populations and seasons during 2013-2018. Multivariate logistic regression model was adopted to fit into the comparisons between non-Norovirus and Norovirus groups. Results: 19.28% of the 12 083 diarrheal cases were found to have carried the Norovirus, with GⅡgroup the most commonly identified genotype. Rates of detection was seen higher in males (20.78%) than in females (17.73%). 30-44 year-old were found having the highest positive rate (21.51%). The positive rates were found the highest (23.60%) in the year of 2015. All the above shown differences were statistically significant (P<0.05). Diarrheal patients affected with Norovirus would present watery stool (75.94%) and vomiting (35.84%). Data from the multivariate logistic analysis showed that factors as: being males, 30-44 years old, officials/clerks, in winter season and with histories of travelling etc., were related to higher risks on Norovirus infection. Conclusions: Infectious diarrhea caused by Norovirus occurred all year round, with seasonal peaks seen in winter and spring, in Shanghai. Specific prevention and control measures should be taken on Norovirus- caused infectious diarrhea, according to the difference on age, gender and season.


Assuntos
Infecções por Caliciviridae/epidemiologia , Diarreia/epidemiologia , Disenteria/epidemiologia , Fezes/virologia , Gastroenterite/epidemiologia , Norovirus/isolamento & purificação , Adulto , Infecções por Caliciviridae/diagnóstico , China/epidemiologia , Diarreia/diagnóstico , Diarreia/virologia , Disenteria/diagnóstico , Disenteria/virologia , Feminino , Gastroenterite/virologia , Humanos , Masculino , Norovirus/genética , Vigilância de Evento Sentinela
18.
Acta Virol ; 63(3): 328-332, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31507200

RESUMO

Acute gastroenteritis (AGE) is one of the most frequently occuring illnesses in children and adults worldwide. In February 2017, two AGE outbreaks occurred in two adjacent schools in Huzhou city, Zhenjiang province of China. We detected high percentages of recombinant norovirus GII.P16/GII.2 in one school and chicken anemia virus (CAV) in another school using next generation sequencing (NGS) and specific PCR. The results highlight the importance of continuous surveillance of GII.P16/GII.2, and suggest the need of further studies on whether CAV causes AGE. Keywords: acute gastroenteritis; norovirus; chicken anemia virus; Huzhou; School.


Assuntos
Infecções por Caliciviridae , Vírus da Anemia da Galinha , Surtos de Doenças , Gastroenterite , Norovirus , Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Criança , China/epidemiologia , Infecções por Circoviridae/epidemiologia , Gastroenterite/diagnóstico , Gastroenterite/epidemiologia , Gastroenterite/virologia , Humanos , Prevalência
19.
J Hosp Infect ; 103(4): 441-446, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31494128

RESUMO

BACKGROUND: The Cepheid® GeneXpert® (GXP) can simultaneously test for norovirus (NV), Clostridium difficile (CD), influenza A/B (IFA/B) and respiratory syncytial virus (RSV). AIM: To compare centralized multiplex polymerase chain reaction (PCR) testing with localized GXP testing at a district general hospital. METHODS: From December 2017 to December 2018, samples received at Whipps Cross University Hospital (WCUH) were first tested at the local laboratory before transport centrally to the Royal London Hospital (RLH). At the RLH, a non-proprietary multiplex reverse transcriptase (RT) PCR assay was performed, which also tested for gastrointestinal or respiratory pathogens not tested for by the GXP. FINDINGS: A total of 1111 stool and respiratory samples were processed at both sites; 591 were respiratory and 520 were stool samples. Compared to centralized testing, the GXP gave sensitivity, specificity, and NPV all in excess of 97%, with the exception of RSV. The RSV assay had a sensitivity of 66.7% (95% confidence interval (CI) 24.1, 94.0) but an NPV of 99.7% (95% CI 98.6, 99.9). At the RLH, 65 (5.9%) additional respiratory or gastrointestinal viruses were detected, predominantly rhinovirus 35 (3.2%) and adenovirus 11 (1.0%). Compared to centralized testing, the median time saved for local respiratory and gastrointestinal sample testing was 19 h and 46 min and 17 h and 6 min, respectively. CONCLUSIONS: Local GXP testing compared to centralized multiplex PCR testing for IF, NV and CD, demonstrated sensitivities, specificities and NPV between 95% and 100%. Turnaround times were faster, enabling quicker infection prevention and control decision making. In our local setting (WCUH), the GXP demonstrated the potential to reduce NV and IFA/B outbreaks.


Assuntos
Infecções por Caliciviridae/diagnóstico , Infecções por Clostridium/diagnóstico , Assistência à Saúde/organização & administração , Influenza Humana/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções por Vírus Respiratório Sincicial/diagnóstico , Pesquisa sobre Serviços de Saúde , Hospitais Gerais , Humanos , Londres , Sensibilidade e Especificidade , Fatores de Tempo
20.
J Hosp Infect ; 103(4): 447-453, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31404566

RESUMO

BACKGROUND: The feasibility of introducing three separate Cepheid GeneXpert® assays was assessed: Xpert SA Nasal Complete, Xpert C. difficile, and Xpert Norovirus for point-of-care testing (POCT) on a ward in a district general hospital. AIM: To establish a seven-day/24 h POCT service for meticillin-resistant Staphylococcus aureus (MRSA), Clostridioides difficile, and norovirus operated solely by healthcare workers (HCWs). METHODS: The Cepheid GeneXpert assays performance characteristics were assessed by comparing the assays to traditional central laboratory methods in terms of clinical turnaround times, hands-on time, number of process steps, time to result and diagnostic accuracy. HCW feedback was collected to consider the potential added value of applying this technology to improve patient flow and clinical care. FINDINGS: In total 1170 tests were carried out over the 16-month study period. The assays significantly reduced hands-on time, process steps, and time to result for identification of all three micro-organisms. Overall agreement with central laboratory testing was >98% for all three assays. Staff members fed back that POCT had a positive impact in terms of clinical utility. CONCLUSION: Xpert SA Nasal Complete for MRSA detection, Xpert C. difficile, and Xpert Norovirus can be used as POCT solely by HCWs in a ward setting. Each assay was used throughout a seven-day/24 h period with potential positive impact on bed management and patient care.


Assuntos
Infecções por Caliciviridae/diagnóstico , Infecções por Clostridium/diagnóstico , Serviços de Diagnóstico/organização & administração , Pessoal de Saúde , Técnicas de Diagnóstico Molecular/métodos , Testes Imediatos , Infecções Estafilocócicas/diagnóstico , /isolamento & purificação , Humanos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Norovirus/isolamento & purificação , Estudos Prospectivos , Fatores de Tempo
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