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  1 / 1010 MEDLINE  
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[PMID]:28455127
[Au] Autor:Camini FC; da Silva Caetano CC; Almeida LT; da Costa Guerra JF; de Mello Silva B; de Queiroz Silva S; de Magalhães JC; de Brito Magalhães CL
[Ad] Endereço:Núcleo de Pesquisas em Ciências Biológicas, NUPEB, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
[Ti] Título:Oxidative stress in Mayaro virus infection.
[So] Source:Virus Res;236:1-8, 2017 05 15.
[Is] ISSN:1872-7492
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Mayaro virus (MAYV) is a neglected tropical arbovirus that causes a febrile syndrome that is sometimes accompanied by incapacitating arthritis/arthralgia. The pathogenesis of MAYV has not been completely defined and oxidative stress mediated by an increase in reactive oxygen species (ROS) and/or depletion of antioxidant defences has been found to contribute to several aspects of viral disease. To investigate whether MAYV induced oxidative stress in host cells, we monitored ROS production, oxidative stress markers and antioxidant defences at different time points after infection. Our results show that MAYV induced significant oxidative stress in infected HepG2 cells, as indicated by the increase of malondialdehyde (MDA) and protein carbonyl levels, and by a significant decrease of the reduced versus oxidized glutathione (GSH/GSSG) ratio. Generally, MAYV-infected HepG2 cells also showed an increase in antioxidant defences. We observed an increase in the superoxide dismutase (SOD) and catalase (CAT) activities and the total glutathione content. To determine whether similar effects occurred in other cell types, we evaluated the ROS, MDA and SOD activity levels in J774 cells after MAYV infection. Similar to our observations in HepG2 cells, the J774 cells showed an increase in ROS, MDA and total SOD activity following MAYV infection. Thus, since the cellular redox environment is influenced by the production and removal of ROS, we hypothesize that the overproduction of ROS was responsible for the oxidative stress in response to the MAYV infection despite the increase in the antioxidant status. This study is the first report on the involvement of oxidative stress during MAYV infection. Collectively, our data shed light on some mechanisms that are operational in host cells following exposure to MAYV.
[Mh] Termos MeSH primário: Infecções por Alphavirus/metabolismo
Alphavirus/fisiologia
Estresse Oxidativo
[Mh] Termos MeSH secundário: Alphavirus/genética
Infecções por Alphavirus/genética
Infecções por Alphavirus/virologia
Catalase/metabolismo
Glutationa/metabolismo
Células Hep G2
Seres Humanos
Malondialdeído/metabolismo
Oxirredução
Espécies Reativas de Oxigênio/metabolismo
Superóxido Dismutase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Reactive Oxygen Species); 4Y8F71G49Q (Malondialdehyde); EC 1.11.1.6 (Catalase); EC 1.15.1.1 (Superoxide Dismutase); GAN16C9B8O (Glutathione)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180226
[Lr] Data última revisão:
180226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170430
[St] Status:MEDLINE


  2 / 1010 MEDLINE  
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[PMID]:28744856
[Au] Autor:Chan YH; Ng LFP
[Ad] Endereço:Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR), Singapore.
[Ti] Título:Age has a role in driving host immunopathological response to alphavirus infection.
[So] Source:Immunology;152(4):545-555, 2017 12.
[Is] ISSN:1365-2567
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Alphaviruses are a group of arthropod-borne pathogens capable of causing a wide spectrum of clinical symptoms, ranging from milder symptoms like rashes, fever and polyarthralgia, to life-threatening encephalitis. This genus of viruses is prevalent globally, and can infect patients across a wide age range. Interestingly, disease severity of virus-infected patients is wide-ranging. Definitions of the pathogenesis of alphaviruses, as well as the host factors influencing disease severity, remain limited. The innate and adaptive immune systems are important host defences against alphavirus infections. Several reports have highlighted the roles of specific immune subsets in contributing to the immune pathogenesis of these viruses. However, immunosenescence, a gradual deterioration of the immune system brought about by the natural advancement of age, affects the functional roles of these immune subsets. This phenomenon compromises the host's ability to defend against alphavirus infection and pathogenesis. In addition, the lack of maturity in the immune system in newborns and infants also results in more severe disease outcomes. In this review, we will summarize the subtle yet diverse physiological changes in the immune system during aging, and how these changes underlie the differences in disease severity for common alphaviruses.
[Mh] Termos MeSH primário: Envelhecimento/imunologia
Infecções por Alphavirus/imunologia
Alphavirus/imunologia
[Mh] Termos MeSH secundário: Fatores Etários
Envelhecimento/patologia
Infecções por Alphavirus/mortalidade
Infecções por Alphavirus/patologia
Animais
Seres Humanos
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Em] Mês de entrada:1711
[Cu] Atualização por classe:180220
[Lr] Data última revisão:
180220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170727
[St] Status:MEDLINE
[do] DOI:10.1111/imm.12799


  3 / 1010 MEDLINE  
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[PMID]:29040304
[Au] Autor:Varjak M; Donald CL; Mottram TJ; Sreenu VB; Merits A; Maringer K; Schnettler E; Kohl A
[Ad] Endereço:MRC-University of Glasgow Centre for Virus Research, Glasgow, Scotland, United Kingdom.
[Ti] Título:Characterization of the Zika virus induced small RNA response in Aedes aegypti cells.
[So] Source:PLoS Negl Trop Dis;11(10):e0006010, 2017 Oct.
[Is] ISSN:1935-2735
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:RNA interference (RNAi) controls arbovirus infections in mosquitoes. Two different RNAi pathways are involved in antiviral responses: the PIWI-interacting RNA (piRNA) and exogenous short interfering RNA (exo-siRNA) pathways, which are characterized by the production of virus-derived small RNAs of 25-29 and 21 nucleotides, respectively. The exo-siRNA pathway is considered to be the key mosquito antiviral response mechanism. In Aedes aegypti-derived cells, Zika virus (ZIKV)-specific siRNAs were produced and loaded into the exo-siRNA pathway effector protein Argonaute 2 (Ago2); although the knockdown of Ago2 did not enhance virus replication. Enhanced ZIKV replication was observed in a Dcr2-knockout cell line suggesting that the exo-siRNA pathway is implicated in the antiviral response. Although ZIKV-specific piRNA-sized small RNAs were detected, these lacked the characteristic piRNA ping-pong signature motif and were bound to Ago3 but not Piwi5 or Piwi6. Silencing of PIWI proteins indicated that the knockdown of Ago3, Piwi5 or Piwi6 did not enhance ZIKV replication and only Piwi4 displayed antiviral activity. We also report that the expression of ZIKV capsid (C) protein amplified the replication of a reporter alphavirus; although, unlike yellow fever virus C protein, it does not inhibit the exo-siRNA pathway. Our findings elucidate ZIKV-mosquito RNAi interactions that are important for understanding its spread.
[Mh] Termos MeSH primário: Aedes/virologia
Interações Hospedeiro-Patógeno
Proteínas de Insetos/genética
Interferência de RNA
RNA Interferente Pequeno
Zika virus/fisiologia
[Mh] Termos MeSH secundário: Aedes/citologia
Alphavirus/genética
Animais
Proteínas do Capsídeo/genética
Linhagem Celular
Proteínas de Insetos/metabolismo
Replicação Viral
Vírus da Febre Amarela/genética
Zika virus/genética
Infecção pelo Zika virus/transmissão
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Capsid Proteins); 0 (Insect Proteins); 0 (RNA, Small Interfering)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171119
[Lr] Data última revisão:
171119
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171018
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pntd.0006010


  4 / 1010 MEDLINE  
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[PMID]:29040262
[Au] Autor:Grossi-Soyster EN; Cook EAJ; de Glanville WA; Thomas LF; Krystosik AR; Lee J; Wamae CN; Kariuki S; Fèvre EM; LaBeaud AD
[Ad] Endereço:Departments of Pediatrics, Infectious Disease Division, Stanford University School of Medicine, Stanford, California, United States of America.
[Ti] Título:Serological and spatial analysis of alphavirus and flavivirus prevalence and risk factors in a rural community in western Kenya.
[So] Source:PLoS Negl Trop Dis;11(10):e0005998, 2017 Oct.
[Is] ISSN:1935-2735
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Alphaviruses, such as chikungunya virus, and flaviviruses, such as dengue virus, are (re)-emerging arboviruses that are endemic in tropical environments. In Africa, arbovirus infections are often undiagnosed and unreported, with febrile illnesses often assumed to be malaria. This cross-sectional study aimed to characterize the seroprevalence of alphaviruses and flaviviruses among children (ages 5-14, n = 250) and adults (ages 15 ≥ 75, n = 250) in western Kenya. Risk factors for seropositivity were explored using Lasso regression. Overall, 67% of participants showed alphavirus seropositivity (CI95 63%-70%), and 1.6% of participants showed flavivirus seropositivity (CI95 0.7%-3%). Children aged 10-14 were more likely to be seropositive to an alphavirus than adults (p < 0.001), suggesting a recent transmission period. Alphavirus and flavivirus seropositivity was detected in the youngest participants (age 5-9), providing evidence of inter-epidemic transmission. Demographic variables that were significantly different amongst those with previous infection versus those without infection included age, education level, and occupation. Behavioral and environmental variables significantly different amongst those in with previous infection to those without infection included taking animals for grazing, fishing, and recent village flooding. Experience of recent fever was also found to be a significant indicator of infection (p = 0.027). These results confirm alphavirus and flavivirus exposure in western Kenya, while illustrating significantly higher alphavirus transmission compared to previous studies.
[Mh] Termos MeSH primário: Infecções por Alphavirus/epidemiologia
Infecções por Alphavirus/virologia
Infecções por Flavivirus/epidemiologia
Infecções por Flavivirus/virologia
População Rural
[Mh] Termos MeSH secundário: Adolescente
Adulto
Idoso
Idoso de 80 Anos ou mais
Alphavirus/classificação
Anticorpos Antivirais/sangue
Criança
Pré-Escolar
Feminino
Flavivirus/classificação
Seres Humanos
Quênia/epidemiologia
Masculino
Meia-Idade
Fatores de Risco
Testes Sorológicos
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Viral)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171110
[Lr] Data última revisão:
171110
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171018
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pntd.0005998


  5 / 1010 MEDLINE  
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[PMID]:28956592
[Au] Autor:Sousa IP; Carvalho CAM; Mendes YS; Weissmuller G; Oliveira AC; Gomes AMO
[Ad] Endereço:Instituto de Bioquímica Médica Leopoldo de Meis, Centro de Ciências da Saúde and ‡Instituto de Biofísica Carlos Chagas Filho, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro , Rio de Janeiro, Rio de Janeiro 21941-902, Brazil.
[Ti] Título:Fusion of a New World Alphavirus with Membrane Microdomains Involving Partially Reversible Conformational Changes in the Viral Spike Proteins.
[So] Source:Biochemistry;56(43):5823-5830, 2017 Oct 31.
[Is] ISSN:1520-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Alphaviruses are enveloped arboviruses mainly proposed to infect host cells by receptor-mediated endocytosis followed by fusion between the viral envelope and the endosomal membrane. The fusion reaction is triggered by low pH and requires the presence of both cholesterol and sphingolipids in the target membrane, suggesting the involvement of lipid rafts in the cell entry mechanism. In this study, we show for the first time the interaction of an enveloped virus with membrane microdomains isolated from living cells. Using Mayaro virus (MAYV), a New World alphavirus, we verified that virus fusion to these domains occurred to a significant extent upon acidification, although its kinetics was quite slow when compared to that of fusion with artificial liposomes demonstrated in a previous work. Surprisingly, when virus was previously exposed to acidic pH, a condition previously shown to inhibit alphavirus binding and fusion to target membranes as well as infectivity, and then reneutralized, its ability to fuse with membrane microdomains at low pH was retained. Interestingly, this observation correlated with a partial reversion of low pH-induced conformational changes in viral proteins and retention of virus infectivity upon reneutralization. Our results suggest that MAYV entry into host cells could alternatively involve internalization via lipid rafts and that the conformational changes triggered by low pH in the viral spike proteins during the entry process are partially reversible.
[Mh] Termos MeSH primário: Alphavirus/química
Lipossomos/química
Fusão de Membrana
Microdomínios da Membrana/química
Proteínas Virais de Fusão/química
Internalização do Vírus
[Mh] Termos MeSH secundário: Alphavirus/metabolismo
Concentração de Íons de Hidrogênio
Microdomínios da Membrana/metabolismo
Proteínas Virais de Fusão/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Liposomes); 0 (Viral Fusion Proteins)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171103
[Lr] Data última revisão:
171103
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170929
[St] Status:MEDLINE
[do] DOI:10.1021/acs.biochem.7b00650


  6 / 1010 MEDLINE  
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[PMID]:28949966
[Au] Autor:Herath TK; Ashby AJ; Jayasuriya NS; Bron JE; Taylor JF; Adams A; Richards RH; Weidmann M; Ferguson HW; Taggart JB; Migaud H; Fordyce MJ; Thompson KD
[Ad] Endereço:Institute of Aquaculture, University of Stirling, Stirling, United Kingdom.
[Ti] Título:Impact of Salmonid alphavirus infection in diploid and triploid Atlantic salmon (Salmo salar L.) fry.
[So] Source:PLoS One;12(9):e0179192, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:With increasing interest in the use of triploid salmon in commercial aquaculture, gaining an understanding of how economically important pathogens affect triploid stocks is important. To compare the susceptibility of diploid and triploid Atlantic salmon (Salmo salar L.) to viral pathogens, fry were experimentally infected with Salmonid alphavirus sub-type 1 (SAV1), the aetiological agent of pancreas disease (PD) affecting Atlantic salmon aquaculture in Europe. Three groups of fry were exposed to the virus via different routes of infection: intraperitoneal injection (IP), bath immersion, or cohabitation (co-hab) and untreated fry were used as a control group. Mortalities commenced in the co-hab challenged diploid and triploid fish from 11 days post infection (dpi), and the experiment was terminated at 17 dpi. Both diploid and triploid IP challenged groups had similar levels of cumulative mortality at the end of the experimental period (41.1% and 38.9% respectively), and these were significantly higher (p < 0.01) than for the other challenge routes. A TaqMan-based quantitative PCR was used to assess SAV load in the heart, a main target organ of the virus, and also liver, which does not normally display any pathological changes during clinical infections, but exhibited severe degenerative lesions in the present study. The median viral RNA copy number was higher in diploid fish compared to triploid fish in both the heart and the liver of all three challenged groups. However, a significant statistical difference (p < 0.05) was only apparent in the liver of the co-hab groups. Diploid fry also displayed significantly higher levels of pancreatic and myocardial degeneration than triploids. This study showed that both diploid and triploid fry are susceptible to experimental SAV1 infection. The lower virus load seen in the triploids compared to the diploids may possibly be related to differences in cell metabolism between the two groups, however, further investigation is necessary to confirm this and also to assess the outcome of PD outbreaks in other developmental stages of the fish when maintained in commercial production systems.
[Mh] Termos MeSH primário: Infecções por Alphavirus/virologia
Diploide
Salmo salar/virologia
Triploidia
[Mh] Termos MeSH secundário: Alphavirus/genética
Animais
Aquicultura
Rim/patologia
Fígado/patologia
Músculo Esquelético/patologia
Miocárdio/patologia
Pâncreas/patologia
RNA Viral/análise
RNA Viral/genética
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Salmo salar/genética
Carga Viral
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171031
[Lr] Data última revisão:
171031
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170927
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0179192


  7 / 1010 MEDLINE  
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[PMID]:28701392
[Au] Autor:Hellström K; Kallio K; Utt A; Quirin T; Jokitalo E; Merits A; Ahola T
[Ad] Endereço:Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.
[Ti] Título:Partially Uncleaved Alphavirus Replicase Forms Spherule Structures in the Presence and Absence of RNA Template.
[So] Source:J Virol;91(18), 2017 Sep 15.
[Is] ISSN:1098-5514
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Alphaviruses are positive-strand RNA viruses expressing their replicase as a polyprotein, P1234, which is cleaved to four final products, nonstructural proteins nsP1 to nsP4. The replicase proteins together with viral RNA and host factors form membrane invaginations termed spherules, which act as the replication complexes producing progeny RNAs. We have previously shown that the wild-type alphavirus replicase requires a functional RNA template and active polymerase to generate spherule structures. However, we now find that specific partially processed forms of the replicase proteins alone can give rise to membrane invaginations in the absence of RNA or replication. The minimal requirement for spherule formation was the expression of properly cleaved nsP4, together with either uncleaved P123 or with the combination of nsP1 and uncleaved P23. These inactive spherules were morphologically less regular than replication-induced spherules. In the presence of template, nsP1 plus uncleaved P23 plus nsP4 could efficiently assemble active replication spherules producing both negative-sense and positive-sense RNA strands. P23 alone did not have membrane affinity, but could be recruited to membrane sites in the presence of nsP1 and nsP4. These results define the set of viral components required for alphavirus replication complex assembly and suggest the possibility that it could be reconstituted from separately expressed nonstructural proteins. All positive-strand RNA viruses extensively modify host cell membranes to serve as efficient platforms for viral RNA replication. Alphaviruses and several other groups induce protective membrane invaginations (spherules) as their genome factories. Most positive-strand viruses produce their replicase as a polyprotein precursor, which is further processed through precise and regulated cleavages. We show here that specific cleavage intermediates of the alphavirus replicase can give rise to spherule structures in the absence of viral RNA. In the presence of template RNA, the same intermediates yield active replication complexes. Thus, partially cleaved replicase proteins play key roles that connect replication complex assembly, membrane deformation, and the different stages of RNA synthesis.
[Mh] Termos MeSH primário: Alphavirus/enzimologia
Interações Hospedeiro-Patógeno
Biogênese de Organelas
Multimerização Proteica
RNA Replicase/metabolismo
Replicação Viral
[Mh] Termos MeSH secundário: Ligação Proteica
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 2.7.7.48 (RNA Replicase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170906
[Lr] Data última revisão:
170906
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170714
[St] Status:MEDLINE


  8 / 1010 MEDLINE  
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[PMID]:28629406
[Au] Autor:Bannai H; Nemoto M; Niwa H; Murakami S; Tsujimura K; Yamanaka T; Kondo T
[Ad] Endereço:Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi, 329-0412, Japan. bannai@equinst.go.jp.
[Ti] Título:Geospatial and temporal associations of Getah virus circulation among pigs and horses around the perimeter of outbreaks in Japanese racehorses in 2014 and 2015.
[So] Source:BMC Vet Res;13(1):187, 2017 Jun 19.
[Is] ISSN:1746-6148
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: We studied a recent epizootic of Getah virus infection among pigs in the southern part of Ibaraki Prefecture and the northern part of Chiba Prefecture, Japan, focusing on its possible association with outbreaks in racehorses in 2014 and 2015. The genomic sequence of a Getah virus strain from an infected pig was analyzed to evaluate the degree of identity with the strains from horses. RESULTS: Sera were collected from pigs from September to December 2012 to 2015 in south Ibaraki (380 pigs in 29 batches), and from September to December 2010 to 2015 in north Chiba (538 pigs in 104 batches). They were examined by using a virus-neutralizing test for Getah virus. Seropositivity rates in 2012-2013 in south Ibaraki and 2010-2012 in north Chiba ranged from 0% to 1.6%. In south Ibaraki, seropositivity rates in 2014 (28.8%) and 2015 (65.0%) were significantly higher than those in the previous years (P < 0.01); 4/5 batches had positive sera in 2014 and 7/7 in 2015. In north Chiba, seropositivity rates in 2013 (14.1%), 2014 (17.8%), and 2015 (48.0%) were significantly higher than those in the previous years (P < 0.01); 6/27 batches had positive sera in 2013, 3/9 in 2014, and 5/5 in 2015. Complete genome analysis revealed that the virus isolated from an infected pig had 99.89% to 99.94% nucleotide identity to the strains isolated from horses during the outbreaks in 2014 and 2015. CONCLUSIONS: Serological surveillance of Getah virus in pigs revealed that the virus was circulating in south Ibaraki and north Chiba in 2014 and 2015; this was concomitant with the outbreaks in racehorses. The Getah virus strain isolated from a pig was closely related to the ones from horses during the 2014 and 2015 outbreaks. To our knowledge, this is the first convincing case of simultaneous circulation of Getah virus both among pigs and horses in specific areas.
[Mh] Termos MeSH primário: Infecções por Alphavirus/veterinária
Alphavirus/isolamento & purificação
Doenças dos Cavalos/virologia
Doenças dos Suínos/virologia
[Mh] Termos MeSH secundário: Alphavirus/classificação
Infecções por Alphavirus/epidemiologia
Infecções por Alphavirus/transmissão
Animais
Cercopithecus aethiops
DNA Viral
Surtos de Doenças/veterinária
Genoma Viral
Doenças dos Cavalos/epidemiologia
Doenças dos Cavalos/transmissão
Cavalos
Japão/epidemiologia
Filogenia
RNA Viral/sangue
Análise de Sequência de DNA
Especificidade da Espécie
Suínos
Doenças dos Suínos/epidemiologia
Doenças dos Suínos/transmissão
Células Vero
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Viral); 0 (RNA, Viral)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170621
[St] Status:MEDLINE
[do] DOI:10.1186/s12917-017-1112-6


  9 / 1010 MEDLINE  
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Vasconcelos, Pedro Fernando da Costa
Texto completo SciELO Brasil
[PMID]:28591313
[Au] Autor:Naveca FG; Nascimento VAD; Souza VC; Nunes BTD; Rodrigues DSG; Vasconcelos PFDC
[Ad] Endereço:Fundação Oswaldo Cruz-Fiocruz, Instituto Leônidas e Maria Deane, Manaus, AM, Brasil.
[Ti] Título:Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses.
[So] Source:Mem Inst Oswaldo Cruz;112(7):510-513, 2017 Jul.
[Is] ISSN:1678-8060
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:We describe a sensitive method for simultaneous detection of Oropouche and Oropouche-like viruses carrying the Oropouche S segment, as well as the Mayaro virus, using a multiplexed one-step reverse transcription real-time polymerase chain reaction (RT-qPCR). A chimeric plasmid containing both Mayaro and Oropouche targets was designed and evaluated for the in vitro production of transcribed RNA, which could be easily used as a non-infectious external control. To track false-negative results due to PCR inhibition or equipment malfunction, the MS2 bacteriophage was also included in the multiplex assay as an internal positive control. The specificity of the multiplex assay was evaluated by Primer-Blast analysis against the entire GenBank database, and further against a panel of 17 RNA arboviruses. The results indicated an accurate and highly sensitive assay with amplification efficiency greater than 98% for both targets, and a limit of detection between two and 20 copies per reaction. We believe that the assay described here will provide a tool for Mayaro and Oropouche virus detection, especially in areas where differential diagnosis of Dengue, Zika and Chikungunya viruses should be performed.
[Mh] Termos MeSH primário: Infecções por Alphavirus/diagnóstico
Alphavirus/genética
Infecções por Bunyaviridae/diagnóstico
Orthobunyavirus/genética
[Mh] Termos MeSH secundário: Alphavirus/classificação
Infecções por Alphavirus/virologia
Infecções por Bunyaviridae/virologia
Seres Humanos
Reação em Cadeia da Polimerase Multiplex
Orthobunyavirus/classificação
Reprodutibilidade dos Testes
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170824
[Lr] Data última revisão:
170824
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170608
[St] Status:MEDLINE


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[PMID]:28500814
[Au] Autor:Hall-Mendelin S; Hewitson GR; Genge D; Burtonclay PJ; De Jong AJ; Pyke AT; van den Hurk AF
[Ad] Endereço:Public Health Virology, Forensic and Scientific Services, Department of Health, Queensland Government, Coopers Plains, Australia.
[Ti] Título:FTA Cards Facilitate Storage, Shipment, and Detection of Arboviruses in Infected Collected in Adult Mosquito Traps.
[So] Source:Am J Trop Med Hyg;96(5):1241-1243, 2017 May.
[Is] ISSN:1476-1645
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:AbstractThe utility of applying infected to Flinders Technology Associates (FTA ) cards for storage, transport, and detection of dengue, Zika, and Barmah Forest viruses was assessed in laboratory-based experiments. The mosquitoes had been removed from Gravid Traps maintained under conditions of high temperature and humidity. RNA of all viruses could be detected in infected mosquitoes on FTA cards either individually or in pools with uninfected mosquitoes, and stored for up to 28 days. Importantly, there was only a minimal decrease in RNA levels in mosquitoes between days 0 and 28, indicating that viral RNA was relatively stable on the cards. FTA cards thus provide a mechanism for storing potentially infected mosquitoes collected in the field and transporting them to a central diagnostic facility for virus detection.
[Mh] Termos MeSH primário: Aedes/virologia
Alphavirus/isolamento & purificação
Arbovirus/isolamento & purificação
Vírus da Dengue/isolamento & purificação
RNA Viral/isolamento & purificação
Fitas Reagentes
Zika virus/isolamento & purificação
[Mh] Termos MeSH secundário: Alphavirus/genética
Animais
Arbovirus/classificação
Arbovirus/genética
Vírus da Dengue/genética
Controle de Mosquitos
Estabilidade de RNA
Reprodutibilidade dos Testes
Manejo de Espécimes/normas
Zika virus/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral); 0 (Reagent Strips)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170803
[Lr] Data última revisão:
170803
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170514
[St] Status:MEDLINE
[do] DOI:10.4269/ajtmh.16-0981



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