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Pesquisa : D13.444.600.223 [Categoria DeCS]
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  1 / 22154 MEDLINE  
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[PMID]:29216103
[Au] Autor:Wu Z; Liao R; Sun X; Zu D; Liu W; Tan H; Sun S
[Ti] Título:Digital quantification of DNA by mapping polarization degree related with coding gold nanorods.
[So] Source:Appl Opt;56(33):9301-9307, 2017 Nov 20.
[Is] ISSN:1539-4522
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The development of highly sensitive and low-cost methods for detecting DNA is of critical importance. Here, we describe a strategy for the highly sensitive and low-cost digital detection of target DNA. Individual DNA molecules were encoded with a single gold nanorod (Au NR), which was then separated and enriched using the magnetic immune-separation process, followed by dehybridization and dispersion into a buffer solution and immobilization on glass slides for polarized dark-field microscopic imaging. With the imaging we can get the first three data sets of the Stokes vector, and the experimental degree of the linear polarization of the light scattered by the Au NR was obtained. Using the Monte Carlo simulation program, the Muller matrix of the Au NRs was simulated and the simulated degree of the linear polarization was calculated to be 0.58. Based on the experimental and simulated degree of the linear polarization, the Au NRs were identified and quantified with an in-house Matlab program, and the concentration of the target DNA at the femtomolar level was therefore achieved.
[Mh] Termos MeSH primário: DNA Viral/análise
Ouro
Vírus da Hepatite B/genética
Luz
Nanotubos
Espalhamento de Radiação
[Mh] Termos MeSH secundário: Sondas de DNA
Microscopia Eletrônica de Varredura
Método de Monte Carlo
Nanotubos/ultraestrutura
Software
Ressonância de Plasmônio de Superfície
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes); 0 (DNA, Viral); 7440-57-5 (Gold)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180206
[Lr] Data última revisão:
180206
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171208
[St] Status:MEDLINE
[do] DOI:10.1364/AO.56.009301


  2 / 22154 MEDLINE  
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[PMID]:29325245
[Au] Autor:Gong JY; Zhang YZ; Zhang JD; Zhang W; Li JQ; Ru K; Liu EB
[Ad] Endereço:Department of Pathology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Tianjin 300020, China.
[Ti] Título:[Clinical characteristics of high-grade B-cell lymphomas with rearrangement of MYC, bcl-6 and bcl-2].
[So] Source:Zhonghua Bing Li Xue Za Zhi;47(1):14-18, 2018 Jan 08.
[Is] ISSN:0529-5807
[Cp] País de publicação:China
[La] Idioma:chi
[Ab] Resumo:To investigate the clinicopathologic features of patients with high-grade B-cell lymphomas (HGBL) that have rearrangements of MYC, bcl-6 and bcl-2. One hundred and fifty-eight B-cell lymphomas patients from Institute of Hematology and Blood Diseases Hospital from January 2016 to April 2017 were detected by fluorescence in situ hybridization (FISH) with double color split-apart probes. Among 158 B-cell lymphomas, 3 cases with MYC, bcl-2 and bcl-6 rearrangements were identified, 1 of which also had CCND1/IgH translocation. All three patients were of older age, with poor prognostic parameters, multiple organs involvements, elevated LDH and advanced-tumor stage. Two of the three patients were treated with high-intensity chemotherapy and had no remission with an overall survival of 9 months and 11 months respectively. One patient had follow-up with no treatment. Histologically, all three cases showed a spectrum of morphologic features. Although initially categorized as lymphoblastic lymphoma, diffuse large lymphoma and mantle cell lymphoma respectively, two cases were associated with germinal center B-cell (GCB) immunophenotype and 1 case with non-GCB immunophenotype. They had a high proliferation index as assessed by immunostaining for Ki-67 (60%-90%). MYC(+) bcl-2(+) bcl-6(+) HGBL is an aggressive disease with multiple organ involvement, high serum LDH levels, advanced stage disease, poor prognosis and shorter patient survival. The diagnosis should be made by histopathology combined with FISH analysis. Its separation from other types of B cell large cell lymphoma is of clinical importance.
[Mh] Termos MeSH primário: Rearranjo Gênico
Genes myc
Linfoma Difuso de Grandes Células B/genética
Proteínas Proto-Oncogênicas c-bcl-2/genética
Proteínas Proto-Oncogênicas c-bcl-6/genética
[Mh] Termos MeSH secundário: Linfócitos B
Ciclina D1/genética
Sondas de DNA
Centro Germinativo
Seres Humanos
Imunofenotipagem
Hibridização in Situ Fluorescente
Linfoma Difuso de Grandes Células B/tratamento farmacológico
Linfoma Difuso de Grandes Células B/patologia
Gradação de Tumores
Prognóstico
Translocação Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (CCND1 protein, human); 0 (DNA Probes); 0 (Proto-Oncogene Proteins c-bcl-2); 0 (Proto-Oncogene Proteins c-bcl-6); 136601-57-5 (Cyclin D1)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180202
[Lr] Data última revisão:
180202
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180112
[St] Status:MEDLINE
[do] DOI:10.3760/cma.j.issn.0529-5807.2018.01.004


  3 / 22154 MEDLINE  
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[PMID]:27775726
[Au] Autor:Shim TS; Estephan ZG; Qian Z; Prosser JH; Lee SY; Chenoweth DM; Lee D; Park SJ; Crocker JC
[Ad] Endereço:Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
[Ti] Título:Shape changing thin films powered by DNA hybridization.
[So] Source:Nat Nanotechnol;12(1):41-47, 2017 01.
[Is] ISSN:1748-3395
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Active materials that respond to physical and chemical stimuli can be used to build dynamic micromachines that lie at the interface between biological systems and engineered devices. In principle, the specific hybridization of DNA can be used to form a library of independent, chemically driven actuators for use in such microrobotic applications and could lead to device capabilities that are not possible with polymer- or metal-layer-based approaches. Here, we report shape changing films that are powered by DNA strand exchange reactions with two different domains that can respond to distinct chemical signals. The films are formed from DNA-grafted gold nanoparticles using a layer-by-layer deposition process. Films consisting of an active and a passive layer show rapid, reversible curling in response to stimulus DNA strands added to solution. Films consisting of two independently addressable active layers display a complex suite of repeatable transformations, involving eight mechanochemical states and incorporating self-righting behaviour.
[Mh] Termos MeSH primário: Sondas de DNA/química
DNA/química
Ouro/química
Membranas Artificiais
Nanopartículas Metálicas/química
Análise de Sequência com Séries de Oligonucleotídeos
[Mh] Termos MeSH secundário: Análise de Sequência com Séries de Oligonucleotídeos/instrumentação
Análise de Sequência com Séries de Oligonucleotídeos/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (DNA Probes); 0 (Membranes, Artificial); 7440-57-5 (Gold); 9007-49-2 (DNA)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180201
[Lr] Data última revisão:
180201
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161108
[St] Status:MEDLINE
[do] DOI:10.1038/nnano.2016.192


  4 / 22154 MEDLINE  
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[PMID]:27771441
[Au] Autor:Wang J; Liu H; Ma C; Wang J; Zhong L; Wu K
[Ad] Endereço:State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410013, China.
[Ti] Título:Label-free monitoring of DNA polymerase activity based on a thrombin-binding aptamer G-quadruplex.
[So] Source:Mol Cell Probes;32:13-17, 2017 04.
[Is] ISSN:1096-1194
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:We have developed a label-free assay for the detection of DNA polymerase activity based on a thrombin-binding aptamer (TBA) G-quadruplex. In the presence of DNA polymerase, the 3'-OH termini of the hairpin substrate are immediately elongated to replace the TBA, which can be recognized quickly by the ThT dye and results in an increase of fluorescence. This method is highly sensitive with a detection limit of 0.1 U/mL. It is simple and cost-effective without any requirement of labeling with a fluorophore-quencher pair. Furthermore, the proposed method can also be applied to analyze the inhibition of DNA polymerase, which clearly indicates that the proposed method can be applied for screening of potential DNA polymerase inhibitors.
[Mh] Termos MeSH primário: Aptâmeros de Nucleotídeos/metabolismo
DNA Polimerase Dirigida por DNA/metabolismo
Quadruplex G
Coloração e Rotulagem
[Mh] Termos MeSH secundário: Sequência de Bases
DNA Polimerase I/metabolismo
Sondas de DNA/metabolismo
Fluorescência
Tiazóis/metabolismo
Fatores de Tempo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Aptamers, Nucleotide); 0 (DNA Probes); 0 (Thiazoles); 145563-68-4 (thrombin aptamer); 2390-54-7 (thioflavin T); EC 2.7.7.- (DNA Polymerase I); EC 2.7.7.7 (DNA-Directed DNA Polymerase)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:180131
[Lr] Data última revisão:
180131
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161026
[St] Status:MEDLINE


  5 / 22154 MEDLINE  
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[PMID]:29048593
[Au] Autor:Krzywkowski T; Nilsson M
[Ad] Endereço:Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, SE-171 65 Solna, Sweden.
[Ti] Título:Fidelity of RNA templated end-joining by chlorella virus DNA ligase and a novel iLock assay with improved direct RNA detection accuracy.
[So] Source:Nucleic Acids Res;45(18):e161, 2017 Oct 13.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Ligation-based nucleic acid detection methods are primarily limited to DNA, since they exhibit poor performance on RNA. This is attributed to reduced end-joining efficiency and/or fidelity of ligases. Interestingly, chlorella virus DNA ligase (PBCV-1 DNA ligase) has recently been shown to possess high RNA-templated DNA end-joining activity; however, its fidelity has not yet been systematically evaluated. Herein, we characterized PBCV-1 ligase for its RNA-templated end-joining fidelity at single base mismatches in 3' and 5' DNA probe termini and found an overall limited end-joining fidelity. To improve the specificity in PBCV-1 ligase-driven RNA detection assays, we utilized structure-specific 5' exonucleolytic activity of Thermus aquaticus DNA polymerase, used in the invader assay. In the iLock (invader padLock) probe assay, padlock probe molecules are activated prior ligation thus the base at the probe ligation junction is read twice in order to aid successful DNA ligation: first, during structure-specific invader cleavage and then during sequence-specific DNA ligation. We report two distinct iLock probe activation mechanisms and systematically evaluate the assay specificity, including single nucleotide polymorphisms on RNA, mRNA and miRNA. We show significant increase in PBCV-1 ligation fidelity in the iLock probe assay configuration for RNA detection.
[Mh] Termos MeSH primário: Técnicas Biossensoriais/métodos
Reparo do DNA por Junção de Extremidades
DNA Ligases/metabolismo
Sondas de DNA/metabolismo
RNA/análise
Moldes Genéticos
Proteínas Virais/metabolismo
[Mh] Termos MeSH secundário: Pareamento Incorreto de Bases/fisiologia
Técnicas Biossensoriais/normas
Sondas de DNA/química
Limite de Detecção
MicroRNAs/genética
MicroRNAs/metabolismo
Polimorfismo de Nucleotídeo Único/fisiologia
RNA/genética
RNA/metabolismo
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
Reação em Cadeia da Polimerase em Tempo Real/métodos
Sensibilidade e Especificidade
Especificidade por Substrato
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes); 0 (MicroRNAs); 0 (RNA, Messenger); 0 (Viral Proteins); 63231-63-0 (RNA); EC 6.5.1.- (Chlorella virus DNA ligase); EC 6.5.1.- (DNA Ligases)
[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:171020
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx708


  6 / 22154 MEDLINE  
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[PMID]:28802177
[Au] Autor:Saeed AA; Sánchez JLA; O'Sullivan CK; Abbas MN
[Ad] Endereço:Electroanal. Lab., Applied Organic Chemistry Department, National Research Centre, El Bohouth St., Dokki, 12622 Giza, Egypt.
[Ti] Título:DNA biosensors based on gold nanoparticles-modified graphene oxide for the detection of breast cancer biomarkers for early diagnosis.
[So] Source:Bioelectrochemistry;118:91-99, 2017 Dec.
[Is] ISSN:1878-562X
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Two different DNA (ERBB2c and CD24c) modified gold nanoparticles and graphene oxide loaded on glassy carbon electrodes were prepared for early detection of breast cancer markers by electrochemical detection of HER2. Comparative study of ERBB2c and CD24c for the detection was carried out. A "sandwich-type" detection strategy was employed in this electrochemical DNA biosensor and its response was measured by amperometric detection. The electrochemical signal enhancement achieved via gold nanoparticles and grapheme oxide system allowed for sensitive detection of the breast cancer biomarker ERBB2 and the control marker CD24. The modified graphene oxide was characterised using Raman spectroscopy, UV-visible spectroscopy, Fourier transform infrared spectroscopy transmission electron microscopy, scanning electron microscopy and energy-dispersive X-ray spectroscopy. The various steps involved in the modification of a glassy carbon electrode with graphene oxide, gold nanoparticles and DNA probes, target and reporter probe were electrochemically characterised using cyclic voltammetry and electrochemical impedance spectroscopy. Using amperometric detection of a horse radish peroxidase label, detection limits of 0.16nM and 0.23nM were obtained with sensitivity 378nA/nM and 219nA/nM for ERBB2 andCD24 respectively.
[Mh] Termos MeSH primário: Biomarcadores Tumorais/análise
Técnicas Biossensoriais/métodos
Neoplasias da Mama/diagnóstico
Sondas de DNA/química
Detecção Precoce de Câncer/métodos
Ouro/química
Grafite/química
[Mh] Termos MeSH secundário: Sequência de Bases
Calibragem
Sondas de DNA/genética
Eletroquímica
Seres Humanos
Nanopartículas Metálicas/química
Hibridização de Ácido Nucleico
Óxidos/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biomarkers, Tumor); 0 (DNA Probes); 0 (Oxides); 7440-57-5 (Gold); 7782-42-5 (Graphite)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170927
[Lr] Data última revisão:
170927
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170813
[St] Status:MEDLINE


  7 / 22154 MEDLINE  
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[PMID]:28800588
[Au] Autor:Strobl F; Ross JA; Stelzer EHK
[Ad] Endereço:Physical Biology / Physikalische Biologie (IZN, FB 15), Buchmann Institute for Molecular Life Sciences (BMLS), Cluster of Excellence Frankfurt-Macromolecular Complexes (CEF-MC), Goethe Universität-Frankfurt am Main (Campus Riedberg), Max-von-Laue-Straße 15, Frankfurt am Main-Germany.
[Ti] Título:Non-lethal genotyping of Tribolium castaneum adults using genomic DNA extracted from wing tissue.
[So] Source:PLoS One;12(8):e0182564, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The red flour beetle Tribolium castaneum has become the second most important insect model organism and is frequently used in developmental biology, genetics and pest-associated research. Consequently, the methodological arsenal increases continuously, but many routinely applied techniques for Drosophila melanogaster and other insect species are still unavailable. For example, a protocol for non-lethal genotyping has not yet been adapted but is particularly useful when individuals with known genotypes are required for downstream experiments. In this study, we present a workflow for non-lethal genotyping of T. castaneum adults based on extracting genomic DNA from wing tissue. In detail, we describe a convenient procedure for wing dissection and a custom method for wing digestion that allows PCR-based genotyping of up to fifty adults in less than an afternoon with a success rate of about 86%. The amount of template is sufficient for up to ten reactions while viability and fertility of the beetles are preserved. We prove the applicability of our protocol by genotyping the white / scarlet gene pair alleles from the black-eyed San Bernadino wild-type and white-eyed Pearl recessive mutant strains spanning four generations. Non-lethal genotyping has the potential to improve and accelerate many workflows: Firstly, during the establishment process of homozygous cultures or during stock keeping of cultures that carry recessively lethal alleles, laborious test crossing is replaced by non-lethal genotyping. Secondly, in genome engineering assays, non-lethal genotyping allows the identification of appropriate founders before they are crossed against wild-types, narrowing the efforts down to only the relevant individuals. Thirdly, non-lethal genotyping simplifies experimental strategies, in which genotype and behavior should be correlated, since the genetic configuration of potential individuals can be determined before the actual behavior assays is performed.
[Mh] Termos MeSH primário: Genótipo
Técnicas de Genotipagem
Tribolium/genética
Asas de Animais/química
[Mh] Termos MeSH secundário: Animais
Sondas de DNA/síntese química
Genes Letais
Reação em Cadeia da Polimerase
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171004
[Lr] Data última revisão:
171004
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170812
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0182564


  8 / 22154 MEDLINE  
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[PMID]:28761341
[Au] Autor:Xia N; Liu K; Zhou Y; Li Y; Yi X
[Ad] Endereço:Key Laboratory of New Optoelectronic Functional Materials, College of Chemistry and Chemical Engineering, Anyang Normal University, Anyang.
[Ti] Título:Sensitive detection of microRNAs based on the conversion of colorimetric assay into electrochemical analysis with duplex-specific nuclease-assisted signal amplification.
[So] Source:Int J Nanomedicine;12:5013-5022, 2017.
[Is] ISSN:1178-2013
[Cp] País de publicação:New Zealand
[La] Idioma:eng
[Ab] Resumo:miRNAs have emerged as new biomarkers for the detection of a wide variety of cancers. By employing duplex-specific nuclease for signal amplification and gold nanoparticles (AuNPs) as the carriers of detection probes, a novel electrochemical assay of miRNAs was performed. The method is based on conversion of the well-known colorimetric assay into electrochemical analysis with enhanced sensitivity. DNA capture probes immobilized on the electrode surface and ferrocene (Fc)-labeled DNA detection probes (denoted "Fc-DNA-Fc") presented in the solution induced the assembly of positively charged AuNPs on the electrode surface through the electrostatic interaction. As a result, a large number of Fc-DNA-Fc molecules were attached on the electrode surface, thus amplifying the electrochemical signal. When duplex-specific nuclease was added to recycle the process of miRNA-initiated digestion of the immobilized DNA probes, Fc-DNA-Fc-induced assembly of AuNPs on the electrode surface could not occur. This resulted in a significant fall in the oxidation current of Fc. The current was found to be inversely proportional to the concentration of miRNAs in the range of 0-25 fM, and a detection limit of 0.1 fM was achieved. Moreover, this work presents a new method for converting colorimetric assays into sensitive electrochemical analyses, and thus would be valuable for design of novel chemical/biosensors.
[Mh] Termos MeSH primário: Colorimetria/métodos
Técnicas Eletroquímicas/métodos
MicroRNAs/análise
Nanopartículas/química
[Mh] Termos MeSH secundário: Técnicas Biossensoriais/métodos
Sondas de DNA/química
Eletrodos
Compostos Ferrosos/química
Ouro/química
Seres Humanos
Limite de Detecção
Metalocenos
MicroRNAs/sangue
MicroRNAs/química
Ácidos Nucleicos Heteroduplexes/química
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes); 0 (Ferrous Compounds); 0 (Metallocenes); 0 (MicroRNAs); 0 (Nucleic Acid Heteroduplexes); 7440-57-5 (Gold); U96PKG90JQ (ferrocene)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170802
[St] Status:MEDLINE
[do] DOI:10.2147/IJN.S138656


  9 / 22154 MEDLINE  
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[PMID]:28737741
[Au] Autor:Rose JC; Stephany JJ; Valente WJ; Trevillian BM; Dang HV; Bielas JH; Maly DJ; Fowler DM
[Ad] Endereço:Department of Chemistry, University of Washington, Seattle, Washington, USA.
[Ti] Título:Rapidly inducible Cas9 and DSB-ddPCR to probe editing kinetics.
[So] Source:Nat Methods;14(9):891-896, 2017 Sep.
[Is] ISSN:1548-7105
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We developed a chemically inducible Cas9 (ciCas9) and a droplet digital PCR assay for double-strand breaks (DSB-ddPCR) to investigate the kinetics of Cas9-mediated generation and repair of DSBs in cells. ciCas9 is a rapidly activated, single-component Cas9 variant engineered by replacing the protein's REC2 domain with the BCL-xL protein and fusing an interacting BH3 peptide to the C terminus. ciCas9 can be tunably activated by a compound that disrupts the BCL-xL-BH3 interaction within minutes. DSB-ddPCR demonstrates time-resolved, highly quantitative, and targeted measurement of DSBs. Combining these tools facilitated an unprecedented exploration of the kinetics of Cas9-mediated DNA cleavage and repair. We find that sgRNAs targeting different sites generally induce cleavage within minutes and repair within 1 or 2 h. However, we observe distinct kinetic profiles, even for proximal sites, and this suggests that target sequence and chromatin state modulate cleavage and repair kinetics.
[Mh] Termos MeSH primário: Caspase 9/genética
Quebras de DNA de Cadeia Dupla
Sondas de DNA/genética
Edição de Genes/métodos
Técnicas de Sonda Molecular
Reação em Cadeia da Polimerase/métodos
[Mh] Termos MeSH secundário: Cinética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes); EC 3.4.22.- (Caspase 9)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:171024
[Lr] Data última revisão:
171024
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170725
[St] Status:MEDLINE
[do] DOI:10.1038/nmeth.4368


  10 / 22154 MEDLINE  
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[PMID]:28719613
[Au] Autor:Pal D; Boby N; Kumar S; Kaur G; Ali SA; Reboud J; Shrivastava S; Gupta PK; Cooper JM; Chaudhuri P
[Ad] Endereço:Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, India.
[Ti] Título:Visual detection of Brucella in bovine biological samples using DNA-activated gold nanoparticles.
[So] Source:PLoS One;12(7):e0180919, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Brucellosis is a bacterial disease, which, although affecting cattle primarily, has been associated with human infections, making its detection an important challenge. The existing gold standard diagnosis relies on the culture of bacteria which is a lengthy and costly process, taking up to 45 days. New technologies based on molecular diagnosis have been proposed, either through dip-stick, immunological assays, which have limited specificity, or using nucleic acid tests, which enable to identify the pathogen, but are impractical for use in the field, where most of the reservoir cases are located. Here we demonstrate a new test based on hybridization assays with metal nanoparticles, which, upon detection of a specific pathogen-derived DNA sequence, yield a visual colour change. We characterise the components used in the assay with a range of analytical techniques and show sensitivities down to 1000 cfu/ml for the detection of Brucella. Finally, we demonstrate that the assay works in a range of bovine samples including semen, milk and urine, opening up the potential for its use in the field, in low-resource settings.
[Mh] Termos MeSH primário: Técnicas Biossensoriais/métodos
Brucella/isolamento & purificação
Sondas de DNA/química
Ouro/química
Nanopartículas Metálicas/química
[Mh] Termos MeSH secundário: Animais
Bovinos
Limite de Detecção
Leite
Hibridização de Ácido Nucleico
Reprodutibilidade dos Testes
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes); 7440-57-5 (Gold)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170925
[Lr] Data última revisão:
170925
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170719
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0180919



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