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2.
Lancet Glob Health ; 9(4): e469-e478, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33740408

RESUMO

BACKGROUND: Lassa fever is a viral haemorrhagic fever endemic in parts of west Africa. New treatments are needed to decrease mortality, but pretrial reference data on the disease characteristics are scarce. We aimed to document baseline characteristics and outcomes for patients hospitalised with Lassa fever in Nigeria. METHODS: We did a prospective cohort study (LASCOPE) at the Federal Medical Centre in Owo, Nigeria. All patients admitted with confirmed Lassa fever were invited to participate and asked to give informed consent. Patients of all ages, including newborn infants, were eligible for inclusion, as were pregnant women. All participants received standard supportive care and intravenous ribavirin according to Nigeria Centre for Disease Control guidelines and underwent systematic biological monitoring for 30 days. Patients' characteristics, care received, mortality, and associated factors were recorded using standard WHO forms. We used univariable and multivariable logistic regression models to investigate an association between baseline characteristics and mortality at day 30. FINDINGS: Between April 5, 2018, and March 15, 2020, 534 patients with confirmed Lassa fever were admitted to hospital, of whom 510 (96%) gave consent and were included in the analysis. The cohort included 258 (51%) male patients, 252 (49%) female patients, 426 (84%) adults, and 84 (16%) children (younger than 18 years). The median time between first symptoms and hospital admission was 8 days (IQR 7-13). At baseline, 176 (38%) of 466 patients had a Lassa fever RT-PCR cycle threshold (Ct) lower than 30. From admission to end of follow-up, 120 (25%) of 484 reached a National Early Warning Score (second version; NEWS2) of 7 or higher, 67 (14%) of 495 reached a Kidney Disease-Improving Global Outcome (KDIGO) stage of 2 or higher, and 41 (8%) of 510 underwent dialysis. All patients received ribavirin for a median of 10 days (IQR 9-13). 62 (12%) patients died (57 [13%] adults and five [6%] children). The median time to death was 3 days (1-6). The baseline factors independently associated with mortality were the following: age 45 years or older (adjusted odds ratio 16·30, 95% CI 5·31-50·30), NEWS2 of 7 or higher (4·79, 1·75-13·10), KDIGO grade 2 or higher (7·52, 2·66-21·20), plasma alanine aminotransferase 3 or more times the upper limit of normal (4·96, 1·69-14·60), and Lassa fever RT-PCR Ct value lower than 30 (4·65, 1·50-14·50). INTERPRETATION: Our findings comprehensively document clinical and biological characteristics of patients with Lassa fever and their relationship with mortality, providing prospective estimates that could be useful for designing future therapeutic trials. Such trials comparing new Lassa fever treatments to a standard of care should take no more than 15% as the reference mortality rate and consider adopting a combination of mortality and need for dialysis as the primary endpoint. FUNDING: Institut National de la Santé et de la Recherche Médicale, University of Oxford, EU, UK Department for International Development, Wellcome Trust, French Ministry of Foreign Affairs, Agence Nationale de Recherches sur le SIDA et les hépatites virales, French National Research Institute for Sustainable Development.


Assuntos
Febre Lassa/mortalidade , Vírus Lassa/isolamento & purificação , Cuidados Paliativos , Ribavirina/administração & dosagem , Administração Intravenosa , Adolescente , Adulto , Fatores Etários , Criança , Pré-Escolar , DNA Viral/isolamento & purificação , Feminino , Seguimentos , Mortalidade Hospitalar , Hospitalização/estatística & dados numéricos , Humanos , Lactente , Recém-Nascido , Estimativa de Kaplan-Meier , Febre Lassa/diagnóstico , Febre Lassa/terapia , Febre Lassa/virologia , Vírus Lassa/genética , Masculino , Pessoa de Meia-Idade , Nigéria/epidemiologia , Gravidez , Prognóstico , Estudos Prospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/estatística & dados numéricos , Fatores de Risco , Índice de Gravidade de Doença , Fatores de Tempo , Resultado do Tratamento , Adulto Jovem
4.
Biotechniques ; 70(3): 149-159, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33512248

RESUMO

One goal of microbial ecology researchers is to capture the maximum amount of information from all organisms in a sample. The recent COVID-19 pandemic, caused by the RNA virus SARS-CoV-2, has highlighted a gap in traditional DNA-based protocols, including the high-throughput methods the authors previously established as field standards. To enable simultaneous SARS-CoV-2 and microbial community profiling, the authors compared the relative performance of two total nucleic acid extraction protocols with the authors' previously benchmarked protocol. The authors included a diverse panel of environmental and host-associated sample types, including body sites commonly swabbed for COVID-19 testing. Here the authors present results comparing the cost, processing time, DNA and RNA yield, microbial community composition, limit of detection and well-to-well contamination between these protocols.


Assuntos
DNA Viral/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Microbiota/genética , RNA Ribossômico 16S/isolamento & purificação , /genética , Animais , Biodiversidade , Gatos , Fracionamento Químico/métodos , Fezes/microbiologia , Fezes/virologia , Feminino , Alimentos e Bebidas Fermentados/microbiologia , Humanos , Limite de Detecção , Masculino , Metagenômica/métodos , Camundongos , Saliva/microbiologia , Saliva/virologia , Pele/microbiologia , Pele/virologia
5.
BMJ Case Rep ; 14(1)2021 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-33431534

RESUMO

A man in his late 30s presented with a several-day history of rectal pain, discharge and bleeding associated with systemic upset. Sexual history revealed receptive anal sex with several male partners in the 2 weeks preceding his clinic visit. Examination of the perianal area was unremarkable. Proctoscopy showed evidence of non-ulcerative proctitis. Microscopy for Gram stain showed pus cells plus extracellular Gram-negative diplococci. The patient was treated for presumptive gonorrhoea and chlamydial infection with ceftriaxone, azithromycin and doxycycline. The patient failed to improve with this treatment regimen. Rectal swab results at 48 hours confirmed the causative agent to be herpes simplex virus (HSV) type 2. The patient was recalled and treated successfully with valaciclovir. This case serves as a useful reminder to clinicians to consider HSV in the differential diagnosis of sexually transmitted proctitis, in the absence of perianal or anorectal ulceration.


Assuntos
Herpes Simples/diagnóstico , Herpesvirus Humano 2/isolamento & purificação , Proctite/diagnóstico , Doenças Sexualmente Transmissíveis/diagnóstico , Adulto , Antivirais/uso terapêutico , DNA Viral/isolamento & purificação , Diagnóstico Diferencial , Gonorreia/diagnóstico , Herpes Simples/tratamento farmacológico , Herpes Simples/transmissão , Herpes Simples/virologia , Herpesvirus Humano 2/genética , Humanos , Doenças Inflamatórias Intestinais/diagnóstico , Mucosa Intestinal/virologia , Masculino , Proctite/tratamento farmacológico , Proctite/virologia , Reto/virologia , Comportamento Sexual , Doenças Sexualmente Transmissíveis/tratamento farmacológico , Doenças Sexualmente Transmissíveis/transmissão , Doenças Sexualmente Transmissíveis/virologia , Valaciclovir/uso terapêutico
6.
J Infect Public Health ; 14(2): 187-192, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33486374

RESUMO

Hepatitis B virus (HBV) is a global human pathogen that can cause life-threatening liver disease including persistent hepatitis, cirrhosis of the liver and hepatocellular cancer. The aim of this present study was to investigate the infection of HBV epidemiology and also examine the HBsAg genotype distribution in the southwestern region of Jizan city in the Saudi Arabia. Since 2015-2018, epidemiological study has been conducted within the city premises of Jazan region. HBsAg genotyping with the molecular biology grade techniques was performed in 50 subjects. In this study, 1888 subjects in the Jazan city of the Southwestern region were diagnosed as positive for HBV. The mean age of the participants was found to be 44.5 ± 16.9 years. Over 68% of males and 31% of females participated in this epidemiological study. Approximately, 88% of Saudi nationalities were recruited with 12% involving non-Saudi subjects Genotyping analysis confirmed that 90% of subjects confirmed genotype-D and 10% confirmed genotypes A, H and E. Anova analysis could find no significant association between the genotype analysis and the clinical data (p > 0.05). The prevalence of HBV in the Jazan region was found to be high based on the clinical data from epidemiological studies conducted in the Southwestern region. Since 2015-2018, a total of 1888 samples have been found to be positive at Jazan premises. The HBsAg genotyping studies confirm in this study 90% of the genotyping-D was documented.


Assuntos
DNA Viral/isolamento & purificação , Vírus da Hepatite B/genética , Hepatite B/epidemiologia , Hepatopatias/epidemiologia , Hepatopatias/virologia , Adulto , Estudos Transversais , DNA Viral/genética , Feminino , Genótipo , Hepatite B/sangue , Hepatite B/diagnóstico , Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Arábia Saudita/epidemiologia
7.
Vet Res Commun ; 45(1): 31-40, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33392909

RESUMO

Gastrointestinal disorders caused by enteric viruses are frequently reported in dogs worldwide, with significant mortality rates in unvaccinated individuals. This study reports the identification and molecular characterization of Canine parvovirus (CPV-2), Canine coronavirus (CcoV), Canine astrovirus (AstV), and Canine calicivirus (CcaV) in a panel of dogs showing severe enteric clinical signs sampled in a typical Mediterranean environment (Sardinia, Italy). At least one of these viral species was detected in 92.3% samples. CPV-2 was the most frequently detected virus (87.2%), followed by AsTv (20.5%), CCoV-IIa (18%), and CCoV-I (10.3%). CCoV-IIb and CaCV were not detected in any sample. Single infection was detected in 24 samples (66.7%), mainly related to CPV-2 (91.7%). Coinfections were present in 33.3% samples with constant detection of CPV-2. Canine coronavirus was present only in coinfected animals. The VP2 sequence analysis of CPV-2 positive samples confirmed the presence of all variants, with CPV-2b most frequently detected. Phylogeny based on the CcoV-IIa spike protein (S) gene allowed to identify 2 different clades among Sardinian isolates but failed to distinguish enteric from pantropic viruses. Study on presence and prevalence of enteroviruses in dogs increase our knowledge about the circulation of these pathogens in the Mediterranean area and highlight the need for dedicated routine vaccine prophylaxis. Molecular analyses of enteric viruses are fundamental to avoid failure of vaccines caused by frequent mutations observed in these enteroviruses.


Assuntos
Infecções por Astroviridae/veterinária , Infecções por Caliciviridae/veterinária , Infecções por Coronavirus/veterinária , Doenças do Cão/virologia , Infecções por Parvoviridae/veterinária , Animais , Astroviridae/genética , Astroviridae/isolamento & purificação , Infecções por Astroviridae/epidemiologia , Infecções por Astroviridae/virologia , Caliciviridae/genética , Caliciviridae/isolamento & purificação , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Coronavirus/genética , Coronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , DNA Viral/isolamento & purificação , Doenças do Cão/epidemiologia , Cães , Fezes/virologia , Feminino , Itália/epidemiologia , Masculino , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Parvovirus/genética , Parvovirus/isolamento & purificação , Filogenia
8.
BMJ Case Rep ; 13(12)2020 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-33370940

RESUMO

A 17-year-old Caucasian male presented to ENT with angular stomatitis, oral ulceration and cervical lymphadenopathy. Over the subsequent 18 months he developed recurrent upper respiratory tract infections, pyrexia of unknown origin, oral ulceration and maxillary sinus osteomyelitis. Extensive investigation ensued from various specialties. Positive investigations included a mild but persistently elevated serum Epstein-Barr virus PCR; however, no unifying diagnosis was elicited. It is noteworthy that a significant factor contributing to a delay in his diagnosis was poor compliance with invasive investigations. Ultimately, deteriorating liver function prompted liver biopsy which confirmed a diagnosis of chronic active Epstein-Barr virus infection (CAEBV). This enabled referral for curative treatment in the form of a stem cell transplant. CAEBV is extremely rare in Western countries. Due to fatal complications early diagnosis is critical for successful treatment. Our case highlights the need for regular clinical re-evaluation and a comprehensive multispecialty approach in such cases.


Assuntos
Infecções por Vírus Epstein-Barr/diagnóstico , Herpesvirus Humano 4/isolamento & purificação , Fígado/patologia , Equipe de Assistência ao Paciente , Adolescente , Biópsia , Doença Crônica/terapia , DNA Viral/sangue , DNA Viral/isolamento & purificação , Infecções por Vírus Epstein-Barr/sangue , Infecções por Vírus Epstein-Barr/terapia , Infecções por Vírus Epstein-Barr/virologia , Herpesvirus Humano 4/genética , Humanos , Fígado/virologia , Testes de Função Hepática , Masculino , Encaminhamento e Consulta , Transplante de Células-Tronco
9.
Artigo em Inglês | MEDLINE | ID: mdl-32899918

RESUMO

The incidence of enteric viruses in treated wastewater and their potential release into the environment or use for agriculture are very critical matters in public health. In our study, PCR (polymerase chain reaction) analysis of enteric viruses was performed on 59 samples of influents and effluents collected from Tubli wastewater treatment plant (Water Pollution Control Center (WPCC)) and Tubli Bay, where the effluents were discharged, in Kingdom of Bahrain during two sampling periods. Four clinically essential waterborne enteric viruses were examined: enterovirus (EV), hepatitis A virus (HAV), astroviruses (AV), and rotaviruses (RV) and compared to standard bacterial and bacteriophages indicators of fecal pollution. Detection rates of EV, AV, HAV, and RV in the influent samples were 100%, 75%, 12.5%, and 12.5%, respectively, while 50% of the effluent samples from Tubli WPCC contained only EV RNA. None of the tested enteric viruses could be detected in any of the samples collected directly from Tubli Bay. Effluent samples from Tubli plant did not show significant seasonal differences. Since detection of enteric viruses genome does not necessarily indicate infectivity, the infectivity of these viruses was evaluated through isolation and growth of indictor bacteria and bacteriophages. High concentration of fecal bacteriological indicators was detected in all effluents samples (100%): 3.20 × 103 cfu/mL for E. coli, 1.32 × 103 cfu/mL for Salmonella spp., and 1.92 × 103 cfu/mL for Shigella spp. E. coli and Salmonella specific bacteriophages were also detected in the effluent samples in high titers. The combined results of PCR and bacterial enumeration point to a probable public health risk via the use of these wastewaters in agriculture or their discharge into the sea. Continuous surveillance of viral and bacterial prevalence and their resistance to sewage disinfection procedures could contribute to a better control of risks associated with the recycling of effluent wastewater and its release into the environment.


Assuntos
DNA Viral/isolamento & purificação , Enterovirus/isolamento & purificação , Escherichia coli/isolamento & purificação , RNA Viral/isolamento & purificação , Esgotos/virologia , Águas Residuárias/microbiologia , Águas Residuárias/virologia , Bactérias/genética , Baías , Enterovirus/genética , Escherichia coli/genética , Humanos , RNA Viral/metabolismo , Vírus/genética , Água , Microbiologia da Água
11.
PLoS One ; 15(7): e0236175, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32697798

RESUMO

Adenoviruses cause upper respiratory infections, conjunctivitis, keratitis, and gastrointestinal illness. These can be fatal in immunocompromised individuals. Adenoviruses have also been engineered into viral vectors to deliver therapeutic genes or induce immunity as vaccine carriers. The success of ocular gene therapy is driven partly by the immunologic and biochemical influences of the intraocular environment. We have shown that versican and hyaluronan modulate adenoviral vector transgene expression through CD44 signaling. Herein we explored the role of these pathways on virus replication and viral protein expression of wild type adenovirus. We report that the addition of vitreous humor (which contains both versican and hyaluronan) increases viral hexon protein levels. Vitreous humor also increased wild type adenovirus DNA replication in vitro. Metalloproteinase and γ-secretase inhibitors, which inhibit CD44 proteolytic activation, blocked adenoviral replication in vitro. Similarly, protein kinase C and RhoA kinase inhibitors, both proteins associated with CD44 mediated pathways, also inhibited wild type adenoviral replication in vitro. Application of metalloproteinase and γ-secretase inhibitors to human conjunctival explants sharply decreased adenoviral vector gene expression. Our results demonstrate that pharmacologic delivery of these inhibitors is easily achievable. The inhibition of these enzymes should be explored as potential therapies of wild type adenoviral infections.


Assuntos
Infecções por Adenoviridae/tratamento farmacológico , Adenoviridae/efeitos dos fármacos , Antivirais/farmacologia , Vetores Genéticos/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Adenoviridae/fisiologia , Infecções por Adenoviridae/virologia , Administração Oftálmica , Amidas/farmacologia , Amidas/uso terapêutico , Antivirais/uso terapêutico , Túnica Conjuntiva/metabolismo , DNA Viral/genética , DNA Viral/isolamento & purificação , Diaminas/farmacologia , Diaminas/uso terapêutico , Dipeptídeos/farmacologia , Dipeptídeos/uso terapêutico , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Vetores Genéticos/fisiologia , Células HeLa , Humanos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/metabolismo , Ácidos Hidroxâmicos/farmacologia , Ácidos Hidroxâmicos/uso terapêutico , Indóis/farmacologia , Indóis/uso terapêutico , Maleimidas/farmacologia , Maleimidas/uso terapêutico , Metaloproteases/antagonistas & inibidores , Metaloproteases/metabolismo , Permeabilidade , Proteína Quinase C/antagonistas & inibidores , Proteína Quinase C/metabolismo , Proteólise/efeitos dos fármacos , Piridinas/farmacologia , Piridinas/uso terapêutico , Transdução de Sinais/efeitos dos fármacos , Tiazóis/farmacologia , Tiazóis/uso terapêutico , Versicanas/metabolismo , Proteínas Virais/genética , Proteínas Virais/metabolismo , Corpo Vítreo/metabolismo , Quinases Associadas a rho/metabolismo
13.
Forensic Sci Med Pathol ; 16(3): 457-462, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32578131

RESUMO

Death due to respiratory infection is commonly encountered at autopsy. With only one opportunity to obtain samples for identification of a causative agent, it is important to ensure that sampling regimes are optimized to provide the greatest detection, without the expense and redundancy that can arise from over-sampling. This study was performed retrospectively using data from Coronial autopsies over the period 2012-2019 from which swabs from the nasopharyngeal region, trachea and lung parenchyma, in addition to samples of lung tissue, had been submitted for multiplex PCR detection of respiratory pathogens. From 97 cases with all four samples, there were 24 with at least one positive result for viral infection. Some cases had multiple positive results and a total of 27 respiratory tract viruses were identified, of which rhinovirus, influenza A virus and respiratory syncytial virus were the most common. Seventeen of the 27 viral infections (63%) were identified in all four samples. However, in nearly all cases (96%) the nasopharyngeal swab detected the infective agent when the multiplex PCR panel had detected infection in any of the four sample types. A nasopharyngeal swab is considered to be an optimal sample for detection of respiratory tract viral infection. As the samples analyzed were acquired before the appearance of the COVID-19 virus, the applicability of this finding for COVID-19 screening is not established.


Assuntos
DNA Viral/isolamento & purificação , Pulmão/virologia , Reação em Cadeia da Polimerase Multiplex , Nasofaringe/virologia , Infecções Respiratórias/diagnóstico , Manejo de Espécimes , Virologia , Viroses/diagnóstico , Vírus/isolamento & purificação , Adulto , Idoso , Idoso de 80 Anos ou mais , Autopsia , Causas de Morte , DNA Viral/classificação , DNA Viral/genética , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Infecções Respiratórias/virologia , Estudos Retrospectivos , Viroses/virologia , Vírus/classificação , Vírus/genética
14.
Small ; 16(32): e2002169, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32578378

RESUMO

The ongoing global novel coronavirus pneumonia COVID-19 outbreak has engendered numerous cases of infection and death. COVID-19 diagnosis relies upon nucleic acid detection; however, currently recommended methods exhibit high false-negative rates and are unable to identify other respiratory virus infections, thereby resulting in patient misdiagnosis and impeding epidemic containment. Combining the advantages of targeted amplification and long-read, real-time nanopore sequencing, herein, nanopore targeted sequencing (NTS) is developed to detect SARS-CoV-2 and other respiratory viruses simultaneously within 6-10 h, with a limit of detection of ten standard plasmid copies per reaction. Compared with its specificity for five common respiratory viruses, the specificity of NTS for SARS-CoV-2 reaches 100%. Parallel testing with approved real-time reverse transcription-polymerase chain reaction kits for SARS-CoV-2 and NTS using 61 nucleic acid samples from suspected COVID-19 cases show that NTS identifies more infected patients (22/61) as positive, while also effectively monitoring for mutated nucleic acid sequences, categorizing types of SARS-CoV-2, and detecting other respiratory viruses in the test sample. NTS is thus suitable for COVID-19 diagnosis; moreover, this platform can be further extended for diagnosing other viruses and pathogens.


Assuntos
Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Nanoporos , Técnicas de Amplificação de Ácido Nucleico/métodos , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , Betacoronavirus/classificação , Infecções por Coronavirus/epidemiologia , DNA Viral/genética , DNA Viral/isolamento & purificação , Genes Virais , Humanos , Limite de Detecção , Mutação , Nanotecnologia , Técnicas de Amplificação de Ácido Nucleico/estatística & dados numéricos , Pandemias , Pneumonia Viral/epidemiologia , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade
15.
Emerg Infect Dis ; 26(9): 2054-2063, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32558639

RESUMO

Since its emergence in Wuhan, China, in December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected ≈6 million persons worldwide. As SARS-CoV-2 spreads across the planet, we explored the range of human cells that can be infected by this virus. We isolated SARS-CoV-2 from 2 infected patients in Toronto, Canada; determined the genomic sequences; and identified single-nucleotide changes in representative populations of our virus stocks. We also tested a wide range of human immune cells for productive infection with SARS-CoV-2. We confirm that human primary peripheral blood mononuclear cells are not permissive for SARS-CoV-2. As SARS-CoV-2 continues to spread globally, it is essential to monitor single-nucleotide polymorphisms in the virus and to continue to isolate circulating viruses to determine viral genotype and phenotype by using in vitro and in vivo infection models.


Assuntos
Betacoronavirus , Infecções por Coronavirus/virologia , Leucócitos Mononucleares/virologia , Pneumonia Viral/virologia , Replicação Viral/genética , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , Betacoronavirus/fisiologia , DNA Viral/genética , DNA Viral/isolamento & purificação , Genótipo , Humanos , Cinética , Pandemias , Polimorfismo de Nucleotídeo Único , Sequenciamento Completo do Genoma
16.
PLoS One ; 15(6): e0235002, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32574197

RESUMO

Streptococcus agalactiae or Group B Streptococcus (GBS) is a leading cause of sepsis in neonates. As a preventative measure prophylactic antibiotic administration is common in pregnant women colonised with GBS, but antibiotic-resistance and adverse effects on neonatal microbiomes may result. Use of bacteriophages (phages) is one option for targeted therapy. To this end, four phages (LF1 -LF4) were isolated from wastewater. They displayed lytic activity in vitro against S. agalactiae isolates collected from pregnant women and neonates, with 190/246 isolates (77.2%) and 10/10 (100%) isolates susceptible to at least one phage, respectively. Phage genomes ranged from 32,205-44,768 bp and all phages were members of the Siphoviridae family. High nucleotide identity (99.9%) was observed between LF1 and LF4, which were closely related to a putative prophage of S. agalactiae. The genome organisation of LF2 differed, and it showed similarity to a different S. agalactiae prophage, while LF3 was more closely related to a Streptococcus pyogenes phage. Lysogenic gene presence (integrase, repressor and regulatory modules), was suggestive of temperate phages. In a therapeutic context, temperate phages are not ideal candidates, however, the broad host range activity of these phages observed on clinical isolates in vitro is promising for future therapeutic approaches including bioengineered phage or lysin applications.


Assuntos
Sepse Neonatal/terapia , Terapia por Fagos , Siphoviridae/genética , Fagos de Streptococcus/genética , Streptococcus agalactiae/virologia , DNA Viral/isolamento & purificação , Feminino , Genômica , Especificidade de Hospedeiro/genética , Humanos , Recém-Nascido , Lisogenia , Sepse Neonatal/microbiologia , Filogenia , Gravidez , Siphoviridae/isolamento & purificação , Fagos de Streptococcus/isolamento & purificação , Streptococcus agalactiae/isolamento & purificação , Streptococcus pyogenes/virologia
17.
Circ Heart Fail ; 13(7): e007103, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32586108

RESUMO

BACKGROUND: Polymerase chain reaction analyses of cardiac tissues have detected viral sequences in up to 67% of cases of myocarditis. However, viruses have not been implicated in giant cell myocarditis (GCM). Furthermore, efforts to detect viruses implicated in myocarditis have been unsuccessful in more accessible samples such as peripheral blood. METHODS: We used Virome Capture Sequencing for Vertbrate Viruses (VirCapSeq-VERT), a method that simultaneously screens for all known vertebrate viruses, to investigate viruses in 33 patients with myocarditis. We investigated peripheral blood mononuclear cells (n=24), plasma (n=27), endomyocardial biopsies (n=2), and cardiac tissue samples from explanted hearts (n=13). RESULTS: Nine patients (27%) had GCM and 4 patients (13%) had fulminant myocarditis. We found the following viruses in the blood of patients with myocarditis: Epstein Barr virus (n=11, 41%), human pegivirus (n=1, 4%), human endogenous retrovirus K (n=27, 100%), and anellovirus (n=15, 56%). All tissue samples from fulminant myocarditis (n=2) and GCM (n=13) contained human endogenous retrovirus K. CONCLUSIONS: No nucleic acids from viruses previously implicated in myocarditis or other human illnesses were detected in relevant amounts in cardiac tissue samples from GCM or in blood samples from other types of myocarditis. These findings do not exclude a role for viral infection in GCM but do suggest that if viruses are implicated, the mechanism is likely to be indirect rather than due to cytotoxic infection of myocardium.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Miocardite/virologia , Viroses/virologia , Vírus/isolamento & purificação , Adulto , Biópsia , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miocardite/patologia , RNA Viral/genética , RNA Viral/isolamento & purificação , Viroses/diagnóstico , Viroses/genética , Vírus/genética
18.
J Occup Health ; 62(1): e12112, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32515872

RESUMO

OBJECTIVE: The aim of the study was to determine and compare the prevalence of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) antibodies and DNA among nurses working in different profiles of healthcare activity. MATERIAL AND METHODS: The study population comprised 120 women (90 exposed healthcare workers and 30 controls). Blood samples were investigated using chemiluminescent microparticle immunoassays (CMIA) tests to detect the presence of EBV VCA IgM, IgG, and CMV IgM, IgG. Plasma CMV and EBV DNA levels were assessed using real-time polymerase chain reaction (PCR). RESULTS: CMV IgG antibodies were present in 87.80% nurses (86.70% in controls), EBV IgG were present in all the nurses studied and in the control group. No statistically significant differences were noted between the subgroups of nurses and the control group as regards IgG CMV, VCA IgG EBV. CMV IgM/EBV IgM antibodies were negative in all the nurses. CMV/EBV DNA was reported only in the study group. It was not found in any of control group participants. CONCLUSIONS: The positive PCR CMV/EBV markers only in the study group can be indicative of the exposure of nurses to these pathogens being greater than in other people not being professionally involved in patient care. In addition, it was observed that the level of CMV IgG antibodies as well as EBV VCA IgG antibodies tended to be linked to the age and the length of work of nurses working in pediatrics.


Assuntos
Anticorpos Antivirais/sangue , Citomegalovirus/imunologia , DNA Viral/isolamento & purificação , Herpesvirus Humano 4/imunologia , Enfermeiras e Enfermeiros , Exposição Ocupacional/análise , Adulto , Idoso , Estudos de Casos e Controles , Infecções por Citomegalovirus/epidemiologia , Infecções por Vírus Epstein-Barr/epidemiologia , Feminino , Humanos , Pessoa de Meia-Idade , Enfermeiras Pediátricas , Transplante de Órgãos , Prevalência , Enfermagem de Atenção Primária
19.
PLoS One ; 15(6): e0234518, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32525936

RESUMO

BACKGROUND/OBJECTIVE: Human papillomavirus (HPV) genotyping and cytology have been recommended for colposcopy triage, but it is unclear which combinations of high-risk HPV (hrHPV) types and cytology with various thresholds provide clinically useful information for the triage after primary HPV screening on self-collected samples. METHOD: Chinese Multi-site Screening Trial (CHIMUST) database focused on self-collected samples was reviewed using the results of Cobas4800 HPV assay. Absolute risks of each genotype for cervical intraepithelial neoplasia 2 or worse/ 3 or worse (CIN2+/CIN3+) were calculated. Triage of atypical squamous cells of undetermined significance (ASCUS) or worse cytology was used as the comparator, and diagnostic accuracy for paired comparisons between algorithms was obtained using McNemar's test. RESULTS: A total of 10, 498 women were included, the overall prevalence of hrHPV, HPV16, HPV18, and Other hrHPV genotypes were 13.7%, 2.4%, 0.8%, and 10.5%, respectively. HPV16-positive women had the highest absolute risk among various genotypes for CIN2+/CIN3+ whether in normal or abnormal cytology (ASCUS or worse) and among all age groups. When compared with the comparator, combining HPV16 positivity and/or high-grade squamous intraepithelial lesion (HSIL) or worse yielded higher specificity (97.7% vs. 97.0%, p<0.0001), similar sensitivity (90.7% vs. 96.3%, p = 0.256) for detection of CIN3+, and a decrease in colposcopy referral rate from 3.5% to 2.7%, similar results were found for CIN2+. Positivity for HPV16 and/or (ASCUS or worse), and positivity for (HPV16 and/or HPV18) and/or (ASCUS or worse) achieved favorable sensitivity compared with the comparator (80.6% and 81.3% vs. 70.1% respectively for CIN2+, p<0.0001; both 96.3% vs. 96.3% for CIN3+, p = 1.000), these algorithms would reduce the colposcopy referral rate to 5.0% and 5.6% respectively, compared with 13.7% of that for HPV alone. CONCLUSIONS: Triage of HPV-positive women on self-collected samples by combining HPV16 or HPV16/18 genotyping with different thresholds of cytology could provide tradeoffs in sensitivity for detecting cervical lesions and colposcopy referral rates, and tailor management in various circumstances of clinical practice.


Assuntos
Técnicas de Genotipagem/métodos , Programas de Rastreamento/métodos , Infecções por Papillomavirus/diagnóstico , Triagem/métodos , Neoplasias do Colo do Útero/diagnóstico , Esfregaço Vaginal/métodos , Adulto , Biópsia/estatística & dados numéricos , Colo do Útero/citologia , Colo do Útero/patologia , Colo do Útero/virologia , Colposcopia/estatística & dados numéricos , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Papillomavirus Humano 16/genética , Papillomavirus Humano 16/isolamento & purificação , Papillomavirus Humano 18/genética , Papillomavirus Humano 18/isolamento & purificação , Humanos , Programas de Rastreamento/estatística & dados numéricos , Pessoa de Meia-Idade , Infecções por Papillomavirus/prevenção & controle , Infecções por Papillomavirus/virologia , Encaminhamento e Consulta/estatística & dados numéricos , Medição de Risco/métodos , Sensibilidade e Especificidade , Manejo de Espécimes/métodos , Triagem/estatística & dados numéricos , Neoplasias do Colo do Útero/prevenção & controle , Neoplasias do Colo do Útero/virologia
20.
PLoS One ; 15(6): e0234438, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32525945

RESUMO

Shiga toxin-producing Escherichia coli (STECs) contamination of produce, as a result of contact with ruminant fecal material, has been associated with serious foodborne illness. Bacteriophages (phages) that infect STECs have primarily been reported to be of cattle origin. However, they likely exist in other environments or in animals that share habitats with cattle, such as goats. To explore the presence and diversity of phages specific to STEC O157 and the top six non-O157 STECs in goat-associated environments, environmental samples consisting of feces (goat and cattle) and soil samples were collected monthly for six months from an organic produce farm. A variety of phages belonging to the Myoviridae, Siphoviridae, and Podoviridae families were isolated from all goat fecal and half of the soil samples. The most commonly isolated phages belonged to Myoviridae and were lytic against STEC O103. The isolated phages had different host ranges, but collectively, showed lytic activity against O157 and the top six non-O157 STEC strains excluding O121. Two non-O157 STECs (O174: H21 and O-antigen-negative: H18) were isolated from soil and cattle feces, respectively. Although prior studies have reported that goats shed STEC into the environment, the findings of the current study suggest that goat feces may also contain lytic STEC-specific phages. The phages of goat origin have the capacity to infect STECs implicated in causing foodborne outbreaks, making them potential candidates for biocontrol pending additional characterization steps. Further work is needed to determine if the addition of goats to the farm environment could potentially reduce the presence of STECs.


Assuntos
Bacteriófagos/isolamento & purificação , Fezes/virologia , Cabras/microbiologia , Escherichia coli Shiga Toxigênica/virologia , Criação de Animais Domésticos/métodos , Animais , Bacteriófagos/genética , California , Bovinos/microbiologia , DNA Viral/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/prevenção & controle , Fazendas , Alimentos Orgânicos/microbiologia , Doenças Transmitidas por Alimentos/microbiologia , Doenças Transmitidas por Alimentos/prevenção & controle , Microbiologia do Solo
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