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[PMID]:29275279
[Au] Autor:Chen J; Park B
[Ad] Endereço:United States Department of Agriculture, Agricultural Research Service, U.S. National Poultry Research Center, 950 College Station Rd, Athens, GA 30605, United States.
[Ti] Título:Effect of immunomagnetic bead size on recovery of foodborne pathogenic bacteria.
[So] Source:Int J Food Microbiol;267:1-8, 2018 Feb 21.
[Is] ISSN:1879-3460
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Immunomagnetic separation (IMS) as a culture-free enrichment sample preparation technique has gained increasing popularity in the development of rapid detection methods for foodborne pathogens. While the use of magnetic nanoparticles in IMS is on the rise due to substantially larger surface area compared to conventional magnetic microparticles, the effects of immunomagnetic bead (IMB) size on pathogen cell recovery are not fully understood. In this study we used IMBs of different sizes (100, 500, and 1000nm diameters) to capture Salmonella Enteritidis, a common foodborne pathogen, from buffer solutions as well as food matrices (chicken carcass rinse and liquid egg white). The IMS recovery and non-specific binding rate were compared. The recoveries of Salmonella cells in buffers was highest using the 100nm IMBs (88-96%), followed by the 500nm (31-89%) and 1000nm (4.1-61%) IMBs, demonstrating a significant size effect. The non-specific binding rates of E. coli also increased as IMB size decreased. A 2-72% reduction in Salmonella recovery was observed in chicken carcass rinse and liquid egg white samples compared to in buffers, and this reduction was more significant using 500 and 1000nm IMBs. However, lower IMS recoveries (10-56%) was found in 100nm IMBs two months after preparation. Overall, magnetic nanoparticles yielded superior IMS efficiency to micrometer size IMBs and were less subjective to interference from food matrices. Nevertheless, their long term stability remains an obstacle towards successful use in IMS.
[Mh] Termos MeSH primário: Escherichia coli/isolamento & purificação
Microbiologia de Alimentos/métodos
Separação Imunomagnética/instrumentação
Separação Imunomagnética/métodos
Tamanho da Partícula
Salmonella enteritidis/isolamento & purificação
[Mh] Termos MeSH secundário: Animais
Galinhas/microbiologia
Clara de Ovo/microbiologia
Microbiologia de Alimentos/normas
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180305
[Lr] Data última revisão:
180305
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171225
[St] Status:MEDLINE


  2 / 3057 MEDLINE  
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[PMID]:29381292
[Au] Autor:Pansini M; Dell'Agli G; Marocco A; Netti PA; Battista E; Lettera V; Vergara P; Allia P; Bonelli B; Tiberto P; Barrera G; Alberto G; Martra G; Arletti R; Esposito S
[Ti] Título:Preparation and Characterization of Magnetic and Porous Metal-Ceramic Nanocomposites from a Zeolite Precursor and Their Application for DNA Separation.
[So] Source:J Biomed Nanotechnol;13(3):337-48, 2017 Mar.
[Is] ISSN:1550-7033
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:In this work, metal-ceramic nanocomposites were obtained through short (up to 2 h) thermal treatments at relatively moderate temperatures (750­800 °C) under a reducing atmosphere, using Fe-exchanged zeolite A as the precursor. The as-obtained materials were characterized by X-ray powder diffraction analysis, N2 adsorption at ­196 °C, and highresolution transmission electron microscopy. The results of these analyses showed that the nanocomposites consisted of a dispersion of metallic Fe nanoparticles within a porous ceramic matrix, mainly based on amorphous silica and alumina. These nanocomposites were magnetically characterized, and their magnetic response was studied. Finally, the obtained metal-ceramic nanocomposite materials were used in the separation of Escherichia coli DNA from a crude cell lysate. The results of the DNA separation experiments showed that the obtained materials could perform this type of separation.
[Mh] Termos MeSH primário: DNA Bacteriano/isolamento & purificação
DNA Bacteriano/efeitos da radiação
Separação Imunomagnética/métodos
Nanocompostos/química
Nanocompostos/ultraestrutura
Ultrafiltração/métodos
Zeolitas/química
[Mh] Termos MeSH secundário: DNA Bacteriano/química
Campos Magnéticos
Teste de Materiais
Ligas Metalo-Cerâmicas/química
Nanocompostos/efeitos da radiação
Nanoporos/ultraestrutura
Tamanho da Partícula
Porosidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Bacterial); 0 (Metal Ceramic Alloys); 1318-02-1 (Zeolites)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180223
[Lr] Data última revisão:
180223
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180131
[St] Status:MEDLINE


  3 / 3057 MEDLINE  
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[PMID]:29237735
[Au] Autor:Dougherty JD
[Ad] Endereço:Departments of Genetics and Psychiatry, Washington University School of Medicine, St. Louis, Missouri 63110 jdougherty@genetics.wustl.edu.
[Ti] Título:The Expanding Toolkit of Translating Ribosome Affinity Purification.
[So] Source:J Neurosci;37(50):12079-12087, 2017 Dec 13.
[Is] ISSN:1529-2401
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Translating ribosome affinity purification is a method initially developed for profiling mRNA from genetically defined cell types in complex tissues. It has been applied both to identify target molecules in cell types that are important for controlling a variety of behaviors in the brain, and to understand the molecular consequences on those cells due to experimental manipulations, ranging from drugs of abuse to disease-causing mutations. Since its inception, a variety of methodological advances are opening new avenues of investigation. These advances include a variety of new methods for targeting cells for translating ribosome affinity purification by features such as their projections or activity, additional tags and mouse reagents increasing the flexibility of the system, and new modifications of the method specifically focused on studying the regulation of translation. The latter includes methods to assess cell type-specific regulation of translation in specific subcellular compartments. Here, I provide a summary of these recent advances and resources, highlighting both new experimental opportunities and areas for future technical development.
[Mh] Termos MeSH primário: Fracionamento Celular/métodos
Perfilação da Expressão Gênica/métodos
Ensaios de Triagem em Larga Escala/métodos
Separação Imunomagnética/métodos
Neuroglia/ultraestrutura
Neurônios/ultraestrutura
Biossíntese de Proteínas
Ribossomos
[Mh] Termos MeSH secundário: Marcadores de Afinidade
Animais
Encéfalo/citologia
Fracionamento Celular/tendências
Linhagem Celular
Cromossomos Artificiais Bacterianos
Dependovirus/genética
Eletroporação
Imunofluorescência
Previsões
Genes Reporter
Vetores Genéticos/genética
Proteínas de Fluorescência Verde/análise
Proteínas de Fluorescência Verde/genética
Camundongos
Neuroglia/metabolismo
Neurônios/classificação
Neurônios/metabolismo
Fases de Leitura Aberta/genética
RNA Mensageiro/genética
RNA Mensageiro/isolamento & purificação
Proteínas Recombinantes de Fusão/análise
Proteínas Ribossômicas/análise
Proteínas Ribossômicas/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Affinity Labels); 0 (RNA, Messenger); 0 (Recombinant Fusion Proteins); 0 (Ribosomal Proteins); 147336-22-9 (Green Fluorescent Proteins)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180130
[Lr] Data última revisão:
180130
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171215
[St] Status:MEDLINE
[do] DOI:10.1523/JNEUROSCI.1929-17.2017


  4 / 3057 MEDLINE  
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[PMID]:28783006
[Au] Autor:Robert C; Huet AC; Suárez-Pantaleón C; Brasseur A; Delahaut P; Gillard N
[Ad] Endereço:a Health Department , CER Groupe , Marloie , Belgium.
[Ti] Título:Development of a confirmatory method for detecting recombinant bovine somatotropin in plasma by immunomagnetic precipitation followed by ultra-high performance liquid chromatography coupled to tandem mass spectrometry.
[So] Source:Food Addit Contam Part A Chem Anal Control Expo Risk Assess;34(11):1925-1934, 2017 Nov.
[Is] ISSN:1944-0057
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Recombinant bovine somatotropin (rbST), a synthetic growth hormone, is used to stimulate growth and enhance milk production in dairy cows. Both its use and the sale of dairy products from treated animals are prohibited in the European Union, as well as in Australia, Canada, Japan, and New Zealand, but authorised in several countries (e.g. Brazil, USA). Screening methods involve detecting anti-rbST antibodies (biomarkers) in treated cows. Confirmatory methods are required to prove rbST abuse. The major challenges in determining rbST are its potentially low levels, its high similarity to native bST, and matrix interferences. To overcome these obstacles, we have developed a method involving immunomagnetic precipitation followed by UHPLC-MS/MS for rbST detection. Briefly, protein G magnetic beads pre-coated with an in-house produced monoclonal antibody were added to plasma. Incubation at room temperature allowed rbST present in the sample to bind to the magnetic beads. After that, magnetic beads were isolated by centrifugation and thoroughly washed (PBS, PBS + 0.2% Tween 20). Finally, rbST was released by alkalinisation and the samples were trypsin digested prior to UHPLC-MS/MS analysis in the MRM mode. Validation was done in accordance with European Commission Decision 2002/657/CE. Matrix-matched calibration with internal standards was used. The decision limit (CCα) reached with this approach was 0.11 µg l .
[Mh] Termos MeSH primário: Hormônio do Crescimento/sangue
Separação Imunomagnética
[Mh] Termos MeSH secundário: Animais
Bovinos
Cromatografia Líquida de Alta Pressão
Proteínas Recombinantes/sangue
Espectrometria de Massas em Tandem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Recombinant Proteins); 9002-72-6 (Growth Hormone)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171016
[Lr] Data última revisão:
171016
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170808
[St] Status:MEDLINE
[do] DOI:10.1080/19440049.2017.1364429


  5 / 3057 MEDLINE  
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[PMID]:28665971
[Au] Autor:Joly P; Schaus T; Sass A; Dienelt A; Cheung AS; Duda GN; Mooney DJ
[Ad] Endereço:Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, United States of America.
[Ti] Título:Biophysical induction of cell release for minimally manipulative cell enrichment strategies.
[So] Source:PLoS One;12(6):e0180568, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The use of autologous cells harvested and subsequently transplanted in an intraoperative environment constitutes a new approach to promote regeneration. Usually cells are isolated by selection methods such as fluorescence- or magnetic- activated cell sorting with residual binding of the antibodies or beads. Thus, cell-based therapies would benefit from the development of new devices for cell isolation that minimally manipulate the target cell population. In the clinic, 5 to 10 percent of fractures do not heal properly and CD31+ cells have been identified as promising candidates to support bone regeneration. The aim of this project was to develop and prototype a simple system to facilitate the enrichment of CD31+ cells from whole blood. After validating the specificity of a commercially available aptamer for CD31, we combined this aptamer with traditional magnetic bead strategies, which led to enrichment of CD31+ cells with a purity of 91±10%. Subsequently, the aptamer was attached to agarose beads (Ø = 100-165 um) that were incorporated into a column-based system to enable capture and subsequent release of the CD31+ enriched cells. Different parameters were investigated to allow a biophysical-based cell release from beads, and a simple mixing was found sufficient to release initially bound cells from the optimized column without the need for any chemicals that promote disassociation. The system led to a significant enrichment of CD31+ cells (initial population: 63±9%, released: 87±3%) with excellent cell viability (released: 97±1%). The composition of the released CD31+ fraction indicated an enrichment of the monocyte population. The angiogenic and osteogenic potential of the released cell population were confirmed in vitro. These results and the simplicity of this system highlight the potential of such approach to enable cell enrichment strategies in intraoperative settings.
[Mh] Termos MeSH primário: Fenômenos Biofísicos
Monócitos/citologia
[Mh] Termos MeSH secundário: Meios de Cultivo Condicionados
Citometria de Fluxo
Células Endoteliais da Veia Umbilical Humana
Seres Humanos
Separação Imunomagnética
Monócitos/imunologia
Neovascularização Fisiológica
Osteogênese
Molécula-1 de Adesão Celular Endotelial de Plaquetas/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; VALIDATION STUDIES
[Nm] Nome de substância:
0 (Culture Media, Conditioned); 0 (Platelet Endothelial Cell Adhesion Molecule-1)
[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:170701
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0180568


  6 / 3057 MEDLINE  
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[PMID]:28652021
[Au] Autor:Li H; Meng QH; Noh H; Batth IS; Somaiah N; Torres KE; Xia X; Wang R; Li S
[Ad] Endereço:Department of Oncology, Affiliated Zhongshan Hospital of Dalian University, Dalian, China. Electronic address: Hli15@mdanderson.org.
[Ti] Título:Detection of circulating tumor cells from cryopreserved human sarcoma peripheral blood mononuclear cells.
[So] Source:Cancer Lett;403:216-223, 2017 Sep 10.
[Is] ISSN:1872-7980
[Cp] País de publicação:Ireland
[La] Idioma:eng
[Ab] Resumo:Circulating tumor cells (CTCs) enter the vasculature or lymphatic system after shedding from the primary tumor. CTCs may serve as "seed" cells for tumor metastasis. The utility of CTCs in clinical applications for sarcoma is not fully investigated, partly owing to the necessity for fresh blood samples and the lack of a CTC-specific antibody. To overcome these drawbacks, we developed a technique for sarcoma CTCs capture and detection using cryopreserved peripheral blood mononuclear cells (PBMCs) and our proprietary cell-surface vimentin (CSV) antibody 84-1, which is specific to tumor cells. This technique was validated by sarcoma cell spiking assay, matched CTCs comparison between fresh and cryopreserved PBMCs, and independent tumor markers in multiple types of sarcoma patient blood samples. The reproducibility was maximized when cryopreserved PBMCs were prepared from fresh blood samples within 2 h of the blood draw. In summary, as far as we are aware, ours is the first report to capture and detect CTCs from cryopreserved PBMCs. Further validation in other types of tumor may help boost the feasibility and utility of CTC-based diagnosis in a centralized laboratory.
[Mh] Termos MeSH primário: Biomarcadores Tumorais/sangue
Neoplasias Ósseas/sangue
Criopreservação
Separação Imunomagnética/métodos
Leucócitos Mononucleares/química
Células Neoplásicas Circulantes/química
Osteossarcoma/sangue
Vimentina/sangue
[Mh] Termos MeSH secundário: Neoplasias Ósseas/patologia
Linhagem Celular Tumoral
Imunofluorescência
Seres Humanos
Leucócitos Mononucleares/patologia
Células Neoplásicas Circulantes/patologia
Osteossarcoma/patologia
Valor Preditivo dos Testes
Prognóstico
Reprodutibilidade dos Testes
Fatores de Tempo
[Pt] Tipo de publicação:JOURNAL ARTICLE; VALIDATION STUDIES
[Nm] Nome de substância:
0 (Biomarkers, Tumor); 0 (Vimentin)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170926
[Lr] Data última revisão:
170926
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170628
[St] Status:MEDLINE


  7 / 3057 MEDLINE  
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[PMID]:28647456
[Au] Autor:Son K; Mukherjee M; McIntyre BAS; Eguez JC; Radford K; LaVigne N; Ethier C; Davoine F; Janssen L; Lacy P; Nair P
[Ad] Endereço:Department of Medicine, McMaster University, Hamilton, Ontario, Canada.
[Ti] Título:Improved recovery of functionally active eosinophils and neutrophils using novel immunomagnetic technology.
[So] Source:J Immunol Methods;449:44-55, 2017 Oct.
[Is] ISSN:1872-7905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Clinically relevant and reliable reports derived from in vitro research are dependent on the choice of cell isolation protocols adopted between different laboratories. Peripheral blood eosinophils are conventionally isolated using density-gradient centrifugation followed by immunomagnetic selection (positive/negative) while neutrophils follow a more simplified dextran-sedimentation methodology. With the increasing sophistication of molecular techniques, methods are now available that promise protocols with reduced user-manipulations, improved efficiency, and better yield without compromising the purity of enriched cell populations. These recent techniques utilize immunomagnetic particles with multiple specificities against differential cell surface markers to negatively select non-target cells from whole blood, greatly reducing the cost/time taken to isolate granulocytes. Herein, we compare the yield efficiencies, purity and baseline activation states of eosinophils/neutrophils isolated using one of these newer protocols that use immunomagnetic beads (MACSxpress isolation) vs. the standard isolation procedures. The study shows that the MACSxpress method consistently allowed higher yields per mL of peripheral blood compared to conventional methods (P<0.001, n=8, Wilcoxon paired test), with high isolation purities for both eosinophils (95.0±1.7%) and neutrophils (94.2±10.1%) assessed by two methods: Wright's staining and flow cytometry. In addition, enumeration of CD63 (marker for eosinophil activation) and CD66b (marker for neutrophil activation) cells within freshly isolated granulocytes, respectively, confirmed that conventional protocols using density-gradient centrifugation caused cellular activation of the granulocytes at baseline compared to the MACSxpress method. In conclusion, MACSxpress isolation kits were found to be superior to conventional techniques for consistent purifications of eosinophils and neutrophils that were suitable for activation assays involving degranulation markers.
[Mh] Termos MeSH primário: Eosinófilos/fisiologia
Separação Imunomagnética/métodos
Neutrófilos/fisiologia
[Mh] Termos MeSH secundário: Centrifugação com Gradiente de Concentração
Citometria de Fluxo
Seres Humanos
Contagem de Leucócitos
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171018
[Lr] Data última revisão:
171018
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170626
[St] Status:MEDLINE


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[PMID]:28551244
[Au] Autor:Zhang Y; Yan C; Yang H; Yu J; Wei H
[Ad] Endereço:Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei 430071, PR China; University of Chinese Academy of Sciences, Beijing 100039, PR China; Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, Schoo
[Ti] Título:Rapid and selective detection of E. coli O157:H7 combining phagomagnetic separation with enzymatic colorimetry.
[So] Source:Food Chem;234:332-338, 2017 Nov 01.
[Is] ISSN:0308-8146
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Mammal IgG antibodies are normally used in conventional immunoassays for E. coli O157:H7, which could lead to false positive results from the presence of protein A producing S. aureus. In this study, a natural specific bacteriophage was isolated and then conjugated with magnetic beads as a capture element in a sandwich format for the rapid and selective detection of E. coli O157:H7. To the best of our knowledge, it was the first time to utilize a natural bacteriophage to develop a phagomagnetic separation combined with colorimetric assay for E. coli O157:H7. The method has an overall time less than 2h with a detection limit of 4.9×10 CFU/mL. No interference from S. aureus was observed. Furthermore, the proposed method was successfully applied to detect E. coli O157:H7 in spiked skim milk. The proposed detection system provided a potential method for E. coli O157:H7 and other pathogenic bacteria in food samples.
[Mh] Termos MeSH primário: Colorimetria/métodos
Escherichia coli O157/isolamento & purificação
Contaminação de Alimentos/análise
Microbiologia de Alimentos/métodos
Separação Imunomagnética/métodos
[Mh] Termos MeSH secundário: Animais
Leite/microbiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170808
[Lr] Data última revisão:
170808
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170529
[St] Status:MEDLINE


  9 / 3057 MEDLINE  
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[PMID]:28530188
[Au] Autor:Busayapongchai P; Siri S
[Ad] Endereço:Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand.
[Ti] Título:Simple assay for screening phytoestrogenic compounds using the oestrogen receptor immobilised magnetite nanoparticles.
[So] Source:IET Nanobiotechnol;11(4):395-402, 2017 Jun.
[Is] ISSN:1751-8741
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:With increasing interests of phytoestrogens for their potential applications, a rapid and simple tool for screening these phytochemicals is still required. In this study, a simple assay to detect phytoestrogens was developed based on the competition binding between the tested samples and the fluorescently labelled oestrogen (E2) to the human ligand binding domain of oestrogen receptor (LBD-ER) that was immobilised on the magnetite nanoparticles (MNPs). The 40-kDa LBD-ER peptide was produced in an system. The synthesised 68.7-nm MNPs were silanised and subsequently covalently linked to the C-terminus of LBD-ER peptide. The LBD-ER immobilised MNPs demonstrated the specific binding for the standard E2 with the equilibrium dissociation constant of 9.56 nM and the binding capacity of 0.08 pmol/1 mg of the MNPs. The LBD-ER immobilised MNPs could evaluate oestrogenic activity of the extracts of and , the reported phytoestrogenic plants, but not progesterone (P4) and extract, the negative controls. The results of this work clearly demonstrated a potential assay for detecting phytoestrogens of crude plant extracts, which is simple and easily adapted to a high throughput format.
[Mh] Termos MeSH primário: Receptor beta de Estrogênio/química
Ensaios de Triagem em Larga Escala/métodos
Separação Imunomagnética/métodos
Nanopartículas de Magnetita/química
Fitoestrógenos/análise
Extratos Vegetais/análise
[Mh] Termos MeSH secundário: Absorção Fisico-Química
Imunoensaio/métodos
Nanopartículas de Magnetita/ultraestrutura
Teste de Materiais
Tamanho da Partícula
Fitoestrógenos/química
Extratos Vegetais/química
Mapeamento de Interação de Proteínas/métodos
Reprodutibilidade dos Testes
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Estrogen Receptor beta); 0 (Magnetite Nanoparticles); 0 (Phytoestrogens); 0 (Plant Extracts)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170817
[Lr] Data última revisão:
170817
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170523
[St] Status:MEDLINE
[do] DOI:10.1049/iet-nbt.2016.0139


  10 / 3057 MEDLINE  
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[PMID]:28499434
[Au] Autor:Stewart LD; Tort N; Meakin P; Argudo JM; Nzuma R; Reid N; Delahay RJ; Ashford R; Montgomery WI; Grant IR
[Ad] Endereço:Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, Northern Ireland, UK.
[Ti] Título:Development of a novel immunochromatographic lateral flow assay specific for Mycobacterium bovis cells and its application in combination with immunomagnetic separation to test badger faeces.
[So] Source:BMC Vet Res;13(1):131, 2017 May 12.
[Is] ISSN:1746-6148
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: The European badger is an important wildlife reservoir of Mycobacterium bovis implicated in the spread of bovine tuberculosis in the United Kingdom and Ireland. Infected badgers are known to shed M. bovis in their urine and faeces, which may contaminate the environment. To aid bovine tuberculosis control efforts novel diagnostic tests for detecting infected and shedding badgers are needed. We proposed development of a novel, rapid immunochromatographic lateral flow device (LFD) as a non-invasive test to detect M. bovis cells in badger faeces. Its application in combination with immunomagnetic separation (IMS) to detect Mycobacterium bovis cells in badger faeces is reported here. RESULTS: A novel prototype LFD for M. bovis cells was successfully developed, with unique specificity for M. bovis and a limit of detection 50% (LOD ) of 1.7 × 10 M. bovis cells/ml. When IMS was employed to selectively capture and concentrate M. bovis cells from badger faeces prior to LFD testing, the LOD of the IMS-LFD assay was 2.8 × 10 M. bovis cells/ml faecal homogenate. Faeces samples collected from latrines at badger setts in a region of endemic bovine tuberculosis infection were tested; 78 (18%) of 441 samples tested IMS-LFD assay positive, whereas 140 (32%) tested IMS-qPCR positive (Kappa agreement -0.009 ± 0.044, p = 0.838). Subsequently, when 130 faeces samples from live captured, or captive, badgers of known infection status (on the basis of StatPak, interferon-γ and/or culture results) were tested, the IMS-LFD assay had higher relative diagnostic specificity (Sp 0.926), but poorer relative diagnostic sensitivity (Se 0.081), than IMS-qPCR (Sp 0.706, Se 0.581) and IMS-culture (Sp 0.794, Se 0.436). CONCLUSIONS: The novel IMS-LFD assay, although very specific for M. bovis, has low analytical sensitivity (indicated by the LOD ) and would only detect badgers shedding high numbers of M. bovis (>10 cells/g) in their faeces. The novel LFD would, therefore, have limited value as a non-invasive test for badger TB surveillance purposes but it may have value for alternative veterinary diagnostic applications.
[Mh] Termos MeSH primário: Fezes/microbiologia
Imunocromatografia/veterinária
Separação Imunomagnética/veterinária
Mustelidae/microbiologia
Mycobacterium bovis/isolamento & purificação
[Mh] Termos MeSH secundário: Animais
Anticorpos Antibacterianos/análise
Separação Imunomagnética/métodos
Sensibilidade e Especificidade
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Bacterial)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170926
[Lr] Data última revisão:
170926
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170514
[St] Status:MEDLINE
[do] DOI:10.1186/s12917-017-1048-x



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