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Registros recuperados: 67
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A glass bead protocol for recovery of host cell free Ehrlichia canis and quantification by Sybr-green real-time PCR Biocell
Cardozo,G. P.; Santos,E. V.; Fachin,A. L.; França,S. C.; Marins,M..
E. canis infection of the canine cell line DH82 is a routine in studies with this bacteria. A protocol for isolation of host cell free bacteria was developed based on the use of glass beads. Improvement of infection with E. canis isolated by this method was detected by real-time PCR.
Tipo: Info:eu-repo/semantics/report Palavras-chave: Ehrlichia canis; Glass bead; Real-time PCR.
Ano: 2011 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S0327-95452011000100005
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A Multiplex real-time PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from Brazilian commercial poultry flocks BJM
Fraga,Aline Padilha; Vargas,Tatiana de; Ikuta,Nilo; Fonseca,André Salvador Kazantzi; Celmer,Álvaro José; Marques,Edmundo Kanan; Lunge,Vagner Ricardo.
Mycoplasma gallisepticum (MS) and Mycoplasma synoviae (MS) are important avian pathogens and cause economic losses to the poultry industry. Molecular biology techniques are currently used for a rapid detection of these pathogens and the adoption of control measures of the diseases. The aim of this study was to develop and validate a technique for simultaneous detection of MG and MS by multiplex real time polymerase chain reaction (PCR). The complete assay (Multiplex MGMS) was designed with primers and probes specific for each pathogen and developed to be carried out in a single tube reaction. Vaccines, MG and MS isolates and DNA from other Mycoplasma species were used for the development and validation of the method. Further, 78 pooled clinical samples...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Real-time PCR; Mycoplasma gallisepticum; Mycoplasma synoviae.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822013000200028
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A pan-European ring trial to validate an International Standard for detection of Vibrio cholerae , Vibrio parahaemolyticus and Vibrio vulnificus in seafoods ArchiMer
Hartnell, R. E.; Stockley, L.; Keay, W.; Rosec, J. -p.; Hervio-heath, Dominique; Van Den Berg, H.; Leoni, F.; Ottaviani, D.; Henigman, U.; Denayer, S.; Serbruyns, B.; Georgsson, F.; Krumova-valcheva, G.; Gyurova, E.; Blanco, C.; Copin, S.; Strauch, E.; Wieczorek, K.; Lopatek, M.; Britova, A.; Hardouin, G.; Lombard, B.; Veld, P. In'T; Leclercq, A.; Baker-austin, C..
Globally, vibrios represent an important and well-established group of bacterial foodborne pathogens. The European Commission (EC) mandated the Comite de European Normalisation (CEN) to undertake work to provide validation data for 15 methods in microbiology to support EC legislation. As part of this mandated work programme, merging of ISO/TS 21872–1:2007, which specifies a horizontal method for the detection of V. parahaemolyticus and V. cholerae, and ISO/TS 21872–2:2007, a similar horizontal method for the detection of potentially pathogenic vibrios other than V. cholerae and V. parahaemolyticus) was proposed. Both parts of ISO/TS 21872 utilized classical culture-based isolation techniques coupled with biochemical confirmation steps. The work also...
Tipo: Text Palavras-chave: Vibrios; Seafood; Prawns; Oysters; Biochemical methods; Real-time PCR; Conventional PCR.
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00425/53680/54525.pdf
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A Rapid and Reliable Method for Molecular Detection of Fusarium guttiforme, the Etiological Agent of Pineapple Fusariosis BABT
Carnielli-Queiroz,Lorena; Fernandes,Patricia Machado Bueno; Fernandes,Antônio Alberto Ribeiro; Ventura,José Aires.
Abstract Pineapple (Ananas comosus var. comosus) fusariosis is an economically important fungal disease affecting the plant and its fruit. A rapid and reliable diagnosis is the base of integrated disease management practices. Fusariosis has resulted in quarantines for pineapple products in Central America, Africa and Asia. Difficulties diagnosing and correctly identifying the fungus Fusarium guttiforme, agent of the pineapple fusariosis, have led to the search for new methodologies, and for this we developed a new reliable molecular method to detect it. For diagnostic purposes, real-time PCR of elongation factor gene 1-α (ef1) was used to rapidly, specifically and sensitively diagnose F. guttiforme. A pathogenicity test was conducted with slips of the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Diagnostic; Diseases; Quarantine; Real-time PCR.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100219
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A real-time PCR genotyping assay to detect FAD2A SNPs in peanuts (Arachis hypogaea L.) Electron. J. Biotechnol.
Barkley,Noelle A; Wang,Ming Li; Pittman,Roy N.
The high oleic (C18:1) phenotype in peanuts has been previously demonstrated to result from a homozygous recessive genotype (ol1ol1ol2ol2) in two homeologous fatty acid desaturase genes (FAD2A and FAD2B) with two key SNPs. These mutant SNPs, specifically G448A in FAD2A and 442insA in FAD2B, significantly limit the normal function of the desaturase enzyme activity which converts oleic acid into linoleic acid by the addition of a second double bond in the hydrocarbon chain. Previously, a genotyping assay was developed to detect wild type and mutant alleles in FAD2B. A real-time PCR assay has now been developed to detect wild type and mutant alleles (G448A) in FAD2A using either seed or leaf tissue. This assay was demonstrated to be applicable for the...
Tipo: Journal article Palavras-chave: Fatty acid composition; Gas chromatography; Peanut (Arachis hypogaea L.); Real-time PCR; SNP genotyping.
Ano: 2011 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000100009
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An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef Ciênc. Tecnol. Aliment.
MERCANOGLU TABAN,Birce; AYTAC,Sait Aykut.
Abstract The aim of this study is to optimize a rapid and specific method for detection of E. coli O157:H7 (EHEC) from food samples with high background flora using an immunomagnetic separation-real-time polymerase chain reaction assay (IMS-RTiPCR). For this, EHEC cells were recovered from raw milk and ground meat samples that were artificially contaminated to the final concentrations of 101, 103 and 105 cfu EHEC/mL or g, after a non-selective pre-enrichment at 35 °C for 8 h following either with or without capturing by micro-sized beads. Then, EHEC cells were identified by RTiPCR. The study was also carried out without any enrichment to evaluate the enrichment efficiency of the assay. By comparing the assay with and without pre-enrichment, we showed that...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Escherichia coli 0157:H7; Immunomagnetic separation; Milk; Raw ground beef; Real-time PCR.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612019000400955
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Assessment of conventional PCR and real-time PCR compared to the gold standard method for screening Streptococcus agalactiae in pregnant women BJID
Ferreira,Michele Berger; de-Paris,Fernanda; Paiva,Rodrigo Minuto; Nunes,Luciana de Souza.
ABSTRACT Group B Streptococcus is a causative agent of invasive neonatal infections. Maternal colonization by Streptococcus agalactiae is a necessary condition for vertical transmission, with efficient screening of pregnant women playing an essential role in the prevention of neonatal infections. In this study, we aimed to compare the performance of conventional polymerase chain reaction and real-time PCR assays as screening methods for S. agalactiae in pregnant women against the microbiological culture method considered as the gold-standard. A total of 130 samples from pregnant women were analyzed for sensitivity, specificity, positive predictive value, and negative predictive value. Statistical analysis was performed using the SPSS software, version...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Streptococcus agalactiae; Pregnant women; Screening; Conventional PCR; Real-time PCR.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702018000600449
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Caractérisation et quantification moléculaires de l’écosystème microbien d’altération du saumon cru et des crevettes cuites ArchiMer
Mace, Sabrina.
Raw salmon and cooked shrimp sold in supermarket are usually found under vacuum or CO2 modified atmosphere packaging (MAP). This type of packaging inhibit growth of some bacteria, therefore, CO2 tolerant bacteria dominate spoilage microbiota of these products. The aim of this study is to improve knowledge about bacteria involved in spoilage. Spoilage microbial ecosystems of raw salmon and tropical cooked shrimp were described using classic miocrobiological and molecular methods like PCR-TTGE and pyrosequencing. Lactic acid bacteria, notably Lactococcus piscium, Carnobacterium maltaromaticum, Carnobacterium divergens, Lactobacillus sakei, and Gram negative bacteria like Photobacterium phosphoreum and Enterobacteriaceae (Serratia spp.) has been described as...
Tipo: Text Palavras-chave: Produits de la mer; Microbiote; PCR-TTGE; Pyroséquençage; Potentiel d’altération; P. phosphoreum; PCR en temps-réel; Seafood products; Microbiota; PCR-TTGE; Pyrosequencing; Spoilage potential; P. phosphoreum; Real-time PCR.
Ano: 2013 URL: https://archimer.ifremer.fr/doc/00166/27773/25963.pdf
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Cellular and molecular responses of haemocytes from Ostrea edulis during in vitro infection by the parasite Bonamia ostreae ArchiMer
Morga, Benjamin; Renault, Tristan; Faury, Nicole; Chollet, Bruno; Arzul, Isabelle.
Bonamia ostreae is a protozoan, affiliated to the order Haplosporidia and to the phylum Cercozoa. This parasite is intracellular and infects haemocytes, cells notably involved in oyster defence mechanisms. Bonamiosis due to the parasite B. ostreae is a disease affecting the flat oyster, Ostrea edulis. The strategies used by protozoan parasites to circumvent host defence mechanisms remain largely unknown in marine bivalve molluscs. In the present work, in vitro experiments were carried out in order to study the interactions between haemocytes from O. edulis and purified parasite. B. ostreae. We monitored cellular and molecular responses of oyster haemocytes by light microscopy, flow cytometry and real-time PCR 1, 2, 4 and 8 h p.i. Light microscopy was used...
Tipo: Text Palavras-chave: Bonamia ostreae; Protozoan; Ostrea edulis; Haemocytes; Real-time PCR; Flow cytometry; Super oxide dismutase.
Ano: 2011 URL: http://archimer.ifremer.fr/doc/00037/14874/18001.pdf
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Comparative analysis of conventional PCR and real-time PCR to diagnose shrimp WSD BJM
Leal,C.A.G.; Carvalho-Castro,G.A.; Cottorello,A.C.; Leite,R.C.; Figueiredo,H.C.P..
The aims of this study were to standard and optimize a qPCR protocol with FAM-BHQ1 probe, and to compare its sensitivity against TaqMan qPCR and PCR methods to diagnose shrimp WSD. The FAM-BHQ1 qPCR presented higher clinical sensitivity and showed to be a robust alternative to detect WSSV in clinical samples.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Clinical samples; Sensitivity; Real-time PCR; WSSV; WSD.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822013000300038
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Comparison of inflammatory microRNA expression in healthy and periodontitis tissues Biocell
Wa Lee,Young H; Am Na,Hee S; Jeong,So Y Eon; Jeong,Sung H Ee; Park,Hae R Youn; Chung,Jin.
MicroRNAs (miRNAs) are short RNA molecules that negatively regulate gene expression primarily by degrading target mRNA or inhibit the translation of protein product. Recently, many reports have shown the altered miRNA expression in various diseases. However, there are no reports on miRNA expression related to periodontitis. Thus, this study aimed to compare the miRNAs differentially expressed in healthy and chronic periodontitis tissues and to determine the miRNAs closely associated with chronic periodontitis. To find out the miRNAs differentially induced in healthy and chronic periodontitis tissues, miRNA microarray was carried out and the expression of miRNAs was confirmed by real-time PCR. According to miRNA microarray analyses, six miRNA genes, let-7a,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Microarray; Real-time PCR; Gingiva.
Ano: 2011 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S0327-95452011000200002
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Detection of genetically modified maize events in Brazilian maize-derived food products Ciênc. Tecnol. Aliment.
Branquinho,Maria Regina; Gomes,Déborah Márcia Vasconcelos; Ferreira,Renata Trotta Barroso; Lawson-Ferreira,Rafael; Cardarelli-Leite,Paola.
The Brazilian government has approved many transgenic maize lines for commercialization and has established a threshold of 1% for food labeling, which underscores need for monitoring programs. Thirty four samples including flours and different types of nacho chips were analyzed by conventional and real-time PCR in 2011 and 2012. The events MON810, Bt11, and TC1507 were detected in most of the samples, and NK603 was present only in the samples analyzed in 2012. The authorized lines GA21, T25, and the unauthorized Bt176 were not detected. All positive samples in the qualitative tests collected in 2011 showed a transgenic content higher than 1%, and none of them was correctly labeled. Regarding the samples collected in 2012, all positive samples were...
Tipo: Info:eu-repo/semantics/article Palavras-chave: GMO; Real-time PCR; Transgenic maize; Food labeling.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612013000300002
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Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit Electron. J. Biotechnol.
Taski-Ajdukovic,Ksenija; Nagl,Nevena; Kovacev,Lazar; Curcic,Zivko; Danojevic,Dario.
Sugar beet is a significant industrial crop, often grown in the areas where summer drought can severely limit root yield and sugar content. In order to improve development of sugar beet cultivars with increased drought tolerance it is necessary to understand plant response to water stress at the genomic level. Since recent research efforts have focused on the molecular response of the plant in order to identify water deficit inducible genes, the aim of this investigation was to develop qRT-PCR methodology for the quantification of gene expression in sugar beet under conditions of water deficiency in vitro. Sugar beet genotypes, selected for different response to water deficit, were grown and multiplied in vitro. Axilary shoots were placed on...
Tipo: Journal article Palavras-chave: Beta vulgaris; Drought; In vitro; Real-time PCR.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000600010
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Development of TaqMan real-time PCR assays for monitoring Vibrio harveyi infection and a plasmid harbored by virulent strains in European abalone Haliotis tuberculata aquaculture ArchiMer
Schikorski, David; Renault, Tristan; Paillard, Christine; Bidault-toffin, A.; Tourbiez, Delphine; Saulnier, Denis.
The Gram-negative bacterium Vibrio harveyi is known to be highly pathogenic for the European abalone Haliotis tuberculata, which is a gastronomically important marine gastropod with a high commercial value. Since 1998, some particular bacterial strains are described as implicated in recurrent mortality outbreaks in French farm and field stocks of abalone. Recently, a 9.6kb plasmid named pVCR1, was shown to be harbored by one highly V. harveyi virulent ORM4 strain suggesting its involvement in virulence phenotype. Thus, we have developed in the present study two TaqMan real-time PCR assays allowing to (i) rapidly and specifically detect, by a duplex procedure and in less than 2 h, both V. harveyi and the presence of plasmid pVCR1 from unidentified bacterial...
Tipo: Text Palavras-chave: Vibrio harveyi; Haliotis tuberculata; Real-time PCR; Plasmid; Marine pathogen; Molecular diagnostic.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00124/23540/21383.pdf
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Diagnóstico direto do complexo Mycobacterium tuberculosis em tecidos de bovinos e bubalinos por nested-PCR. Infoteca-e
ARAÚJO, C. P. de; ARAUJO, F. R..
2015
Tipo: Boletim de Pesquisa e Desenvolvimento (INFOTECA-E) Palavras-chave: Tuberculose bovina e bubalina; Nested-PCR; Real-time PCR; Tecido; Inspeção sanitária.
Ano: 2015 URL: http://www.infoteca.cnptia.embrapa.br/handle/doc/1029171
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Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR Repositório Alice
ARAÚJO, C. P.; OSÓRIO, A. L. A. R.; JORGE, K. S. G.; RAMOS, C. A. N.; SOUZA FILHO, A. F.; VIDAL, C. E. S.; VARGAS, A. P. C.; ROXO, E; ROCHA, A. S.; SUFFYS, P. N.; FONSECA JÚNIOR, A. A.; SILVA, M. R.; BARBOSA NETO, J. D.; CERQUEIRA, V. D.; ARAUJO, F. R..
2014
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: Bovine and bubaline tuberculosis; Nested-PCR; Real-time PCR; Tissue; Sanitary inspection; Bovine and bubaline tuberculosis; Nested-PCR; Real-time PCR; Sanitary inspec - tion; Tissue.
Ano: 2014 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1010353
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Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR BJM
Araújo,Cristina P.; Osório,Ana Luiza A.R.; Jorge,Klaudia S.G.; Ramos,Carlos A.N.; Souza Filho,Antonio F.; Vidal,Carlos E.S.; Vargas,Agueda P.C.; Roxo,Eliana; Rocha,Adalgiza S.; Suffys,Philip N.; Fonseca Júnior,Antônio A.; Silva,Marcio R.; Barbosa Neto,José D.; Cerqueira,Valíria D.; Araújo,Flábio R..
Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine and bubaline tuberculosis; Nested-PCR; Real-time PCR; Tissue; Sanitary inspection.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200035
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Duplex realtime PCR method for Epstein-Barr virus and human DNA quantification: its application for post-transplant lymphoproliferative disorders detection BJID
Fellner,María Dolores; Durand,Karina; Rodriguez,Marcelo; Irazu,Lucía; Alonio,Virginia; Picconi,María Alejandra.
INTRODUCTION: The quantification of circulating Epstein-Barr virus (EBV) DNA is used to monitor transplant patients as an early marker of Post-Transplant Lymphoproliferative Disorders (PTLD). So far no standardized methodology exists for such determination.OBJECTIVE: Our purpose was to develop and validate a real-time PCR assay to quantify EBV DNA in clinical samples from transplant recipients.METHODS: A duplex real-time PCR method was developed to amplify DNA from EBV and from a human gene. The EBV load was determined in peripheral blood mononuclear cells (PBMC), plasma and oropharyngeal tissue from 64 non-transplanted patients with lymphoid-hypertrophy (Non-Tx), 47 transplant recipients without PTLD (Tx), 54 recipients with PTLD (Tx-PTLD), and 66 blood...
Tipo: Info:eu-repo/semantics/article Palavras-chave: EBV; Real-time PCR; Viral load; PTLD.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702014000300271
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Early molecular markers predictive of dengue hemorrhagic fever Anais da ABC (AABC)
Calzavara-Silva,Carlos E.; Gomes,Ana L.V.; Maia,Rita C.C.; Acioli-Santos,Bartolomeu; Gil,Laura H.V.G.; Marques Jr.,Ernesto T.A..
The management of acute dengue patients during outbreaks is a challenging problem. Most of the dengue fever cases are benign, but some cases develop into a severe and possibly lethal vasculopathy, known as dengue hemorrhagic fever. Early symptoms of dengue and hemorrhagic fever are very similar. An early differential diagnosis is needed to predict which of these two clinical presentations is crucial to proper patient care and public health management. This study evaluates the predictive potential of specific mRNA expression markers of dengue hemorrhagic fever using quantitative real-time PCR assays. Six candidate "dengue hemorrhagic fever specific signature genes" were evaluated and all showed good correlation among their transcription levels at early days...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Real-time PCR; Dengue hemorrhagic fever; Molecular markers.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652009000400006
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Effect of cryopreservation on the efficiency of exogenous gene, genetic transformation and expression level of Arabidopsis thaliana Electron. J. Biotechnol.
Li,Zhongai; Du,Yaqiong; Wang,Zicheng.
Background: Cryopreservation refers to the storage of a living organism at ultra-low-temperature for long-term preservation of plant germplasm. The effect of cryopreservation on the efficiency of exogenous gene genetic transformation and expression level were studied herein. In this work, transgenic Arabidopsis thaliana were successfully conserved in vitro by cryopreservation methods. Results: The effects of osmotic stress due to cryoprotectants during pretreatment and of storage at -196ºC on the stability, the efficiency of genetic transformation and the expression level of exogenous gene were analyzed in Arabidopsis. The results showed that there had not any significant increasing in the efficiency of genetic transformation after cryopreservation, and...
Tipo: Journal article Palavras-chave: Arabidopsis thaliana; Cryopreservation; Real-time PCR; Transformation efficiency.
Ano: 2013 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600012
Registros recuperados: 67
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