Consulta: subjectFacets:"Real time PCR"
Registros recuperados: 28
Data/hora: 17/05/2024 00:43:15

A cost-effective melting temperature assay for the detection of single-nucleotide polymorphism in the MBL2 gene of HIV-1-infected children
Provedor de dados: 56
Autores: Arraes,L.C.; de Souza,P.R.; Bruneska,D.; Castelo Filho,A.; de Souza Cavada,B.; de Lima Filho,J.L.; Crovella,S.
We report a fast (less than 3 h) and cost-effective melting temperature assay method for the detection of single-nucleotide polymorphisms in the MBL2 gene. The protocol, which is based on the Corbett Rotor Gene real time PCR platform and SYBR Green I chemistry, yielded, in the cohorts studied, sensitive (100%) and specific (100%) PCR amplification without the use of costly fluorophore-labeled probes or post-PCR manipulation. At the end of the PCR, the dissociation protocol included a slow heating from 60º to 95ºC in 0.2ºC steps, with an 8-s interval between steps. Melting curve profiles were obtained using the dissociation software of the Rotor Gene-3000 apparatus. Samples were analyzed in duplicate and in different PCR runs to test the reproducibility of...
Tipo: Info:eu-repo/semantics/other
Palavras-chave: Melting temperature assay; Real time PCR; MBL2; Polymorphisms; HIV-1-positive children.
Ano: 2006
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000600003

A relationship between antimicrobial peptide gene expression and capacity of a selected shrimp line to survive a Vibrio infection
Provedor de dados: 5
Autores: De Lorgeril, Julien; Gueguen, Yannick; Goarant, Cyrille; Goyard, Emmanuel; Mugnier, Chantal; Fievet, Julie; Piquemal, D; Bachere, Evelyne.
Understanding of antimicrobial defence mechanisms of penaeid shrimp should help in the design of efficient strategies for the management and disease control in aquaculture. In this study, we have specifically analysed the expression in circulating hemocytes of antimicrobial peptides (AMPs) encoding genes, such as PEN2 and PEN3, ALF, crustin, lysozyme and a putative cysteine-rich peptide. We evidenced a relationship between the level of expression of some AMPs and the successful response of the shrimp, Litopenaeus stylirostris, to circumvent a pathogenic Vibrio penaeicida infection. Additionally, significant differences in some AMP transcript amounts are evidenced between control, non-selected shrimp line and the third generation breeding of shrimp selected...
Tipo: Text
Palavras-chave: Real time PCR; Cysteine rich peptide; Crustin; Anti LPS factor; Lysozyme; Penaeidins; Immune response; Vibrio penaeicida; Penaeid; Decapoda; Crustacean.
Ano: 2008
URL: http://archimer.ifremer.fr/doc/2008/publication-4524.pdf

Analysis of gene expression in Kalanchoe daigremontiana leaves during plantlet formation under drought stress
Provedor de dados: 69
Autores: Zhong,Tianxiu; Zhu,Chen; Zeng,Huiming; Han,Liebao.
Background: Kalanchoe daigremontiana is an attractive model system for the study of the molecular mechanisms of somatic embryogenesis and organogenesis competence due to its formation of plantlets with adventitious roots on the leaf margins that are derived from somatic embryos. The suppression subtractive hybridization technique was used to investigate gene expression during asexual reproduction. Leaves from plants subjected to drought stress provided the source of &amp;#8216;Tester&amp;#8217; DNA, and leaves from plants grown under normal conditions provided the &amp;#8216;Driver&amp;#8217; DNA for subtractive hybridization. Results: A total of 481 high quality ESTs were generated, which clustered into 390 unigenes. Of these unigenes, 132 grouped into 12...
Tipo: Journal article
Palavras-chave: Bioinformatics analysis; Real time PCR; Suppression subtractive hybridization.
Ano: 2013
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600004

Comparative Analysis of C-repeat Binding Factors (CBFs) in Tomato and Arabidopsis
Provedor de dados: 52
Autores: Heidari,Parviz.
Abstract Cold stress is one of the limiting factors of plant production that plants use different mechanisms for cold tolerance. CBF genes play critical role to regulate the cold responsive genes. To better understand of CBF gene functions, the tomato-CBFs and Arabidopsis-CBFs were evaluated using bioinformatics tools, and finally the expression patterns of SlCBF1 gene were analyzed under 10 and 4˚C in two contrasting tomato species (Solanum lycopersicum and S. habrochaites). The different cis regulatory elements were observed in promoter region of SlCBF1 and AtCBF1 genes, and ICE1, COR and HOS1 proteins exhibited high interaction with CBFs. The results of Real time PCR of SlCBF1 exhibited that under 10 and 4 ˚C, SlCBF1 was down regulated in cold sensitive...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Cold stress; CBF genes; Real time PCR; Tomato; Bioinformatics analysis.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100208

Comparison of nine DNA extraction methods for the diagnosis of bovine tuberculosis by real time PCR
Provedor de dados: 65
Autores: Moura,André; Hodon,Mikael Arrais; Soares Filho,Paulo Martins; Issa,Marina de Azevedo; Oliveira,Ana Paula Ferreira de; Fonseca Júnior,Antônio Augusto.
ABSTRACT: Bovine tuberculosis is an infectious disease with a high impact on the cattle industry, particularly in developing countries. PCR is a very sensitive method for detection of infectious agents, but the sensitivity of molecular diagnosis is largely dependent on the efficiency of the DNA extraction methods. The objective of this study was to evaluate DNA extraction methods for direct detection of  Mycobacterium bovis in bovine tissue. Nine commercial kits for DNA extraction were evaluated when combined with two real time PCRs. The DNeasy Blood &amp; Tissue Kit from QIAGEN showed better performance and sensitivity followed by the DNA Mini Kit RBC and FTA Elute Micro Card. Results suggested that, even when the analytical sensitivity of the qPCR is...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Bovine tuberculosis; DNA extraction; Real time PCR.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016000701223

Detección de Erwinia  chrysanthemi pv zeae (Sabet 1954) Victoria et al., 1975 en maíz en Morelos, México.
Provedor de dados: 32
Autores: Durán Peralta, Elisa.
La “pudrición del tallo del maíz” causada por Erwinia chrysanthemi pv zeae es una de las principales enfermedades del maíz (Zea mays L.) en los países tropicales y subtropicales. En el presente trabajo se realizó la  extracción  de ADN del patógeno y se realizó PCR  utilizando los iniciadores ADE1 y ADE2, diseñados para E. chrysanthemi (Burkh.) Young et al. 1978,   que amplifican un fragmento de 420pb del gen pelADE. Se demostró que estos también pueden ser utilizados para su detección por PCR en tiempo real con resultados similares a los citados en la literatura; sin embargo no pueden ser empleados para la cuantificación de la bacteria debido a la formación de estructuras secundarias. Mediante la caracterización fisiológica, bioquímica y análisis...
Palavras-chave: Zea mays L.; Erwinia chrysanthemi pv zeae; Pudrición del tallo del maíz; PCR en tiempo real; Gen pe/ADE; Corn stalk rot; Real time PCR; Gene Pe/ADE; Fitopatología; Maestría.
Ano: 2011
URL: http://hdl.handle.net/10521/555

Detección de Erwinia  chrysanthemi pv zeae (Sabet 1954) Victoria et al., 1975 en maíz en Morelos, México.
Provedor de dados: 32
Autores: Durán Peralta, Elisa.
La “pudrición del tallo del maíz” causada por Erwinia chrysanthemi pv zeae es una de las principales enfermedades del maíz (Zea mays L.) en los países tropicales y subtropicales. En el presente trabajo se realizó la  extracción  de ADN del patógeno y se realizó PCR  utilizando los iniciadores ADE1 y ADE2, diseñados para E. chrysanthemi (Burkh.) Young et al. 1978,   que amplifican un fragmento de 420pb del gen pelADE. Se demostró que estos también pueden ser utilizados para su detección por PCR en tiempo real con resultados similares a los citados en la literatura; sin embargo no pueden ser empleados para la cuantificación de la bacteria debido a la formación de estructuras secundarias. Mediante la caracterización fisiológica, bioquímica y análisis...
Palavras-chave: Zea mays L.; Erwinia chrysanthemi pv zeae; Pudrición del tallo del maíz; PCR en tiempo real; Gen pe/ADE; Corn stalk rot; Real time PCR; Gene Pe/ADE; Fitopatología; Maestría.
Ano: 2011
URL: http://hdl.handle.net/10521/555

Different methods of real-time PCR for detection of pseudorabies virus
Provedor de dados: 65
Autores: Nonaka,Carolina Kymie Vasques; Fonseca Junior,Antônio Augusto; Guedes,Estefânia Oliveira; D´Ambros,Régia Maria; Lima,Graciela Kunrath; Camargos,Marcelo Fernandes; Heinemann,Marcos Bryan.
ABSTRACT: Pseudorabies (PR) is a highly contagious viral disease of great animal health and economic importance in swine industry. The aim of this study was to evaluate different genomic regions, real-time PCR chemistries and equipment for the molecular diagnosis of PR. Eight primer pairs targeting four genes (gB, gC, gE, gD), three different qPCR chemistries (SybrGreen, hydrolysis probes and plexor) and two equipment (ABI7500, Rotorgene 3000) were evaluated. Oligonucleotides targeting gB using hydrolysis probes showed the best performance after evaluating efficiency (99%), the detection limit (10-1.5 TCID50 mL-1) and diagnostic sensitivity and; therefore, those primers were selected for performance verification factors such as repeatability,...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Real time PCR; Pseudorabies; Diagnose.
Ano: 2017
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782017000300452

Differential gene expression and mitotic cell analysis of the drought tolerant soybean (Glycine max L. Merrill Fabales, Fabaceae) cultivar MG/BR46 (Conquista) under two water deficit induction systems
Provedor de dados: 74
Autores: Martins,Polyana K.; Jordão,Berenice Q.; Yamanaka,Naoki; Farias,José R.B.; Beneventi,Magda A.; Binneck,Eliseu; Fuganti,Renata; Stolf,Renata; Nepomuceno,Alexandre L.
Drought cause serious yield losses in soybean (Glycine max), roots being the first plant organ to detect the water-stress signals triggering defense mechanisms. We used two drought induction systems to identify genes differentially expressed in the roots of the drought-tolerant soybean cultivar MG/BR46 (Conquista) and characterize their expression levels during water deficit. Soybean plants grown in nutrient solution hydroponically and in sand-pots were submitted to water stress and gene expression analysis was conducted using the differential display (DD) and real time polymerase chain reaction (PCR) techniques. Three differentially expressed mRNA transcripts showed homology to the Antirrhinum majus basic helix-loop-helix transcription factor bHLH, the...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Cell division; Differential display; Gene expression; Real time PCR; Soybean; Water stress.
Ano: 2008
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000300019

Early remodeling of rat cardiac muscle induced by swimming training
Provedor de dados: 56
Autores: Verzola,R.M.M.; Mesquita,R.A.; Peviani,S.; Ramos,O.H.P.; Moriscot,A.S.; Perez,S.E.A.; Selistre-de-Araújo,H.S.
The aim of the present investigation was to study the effect of acute swimming training with an anaerobic component on matrix metallopeptidase (MMP) activity and myosin heavy chain gene expression in the rat myocardium. Animals (male Wistar rats, weighing approximately 180 g) were trained for 6 h/day in 3 sessions of 2 h each for 1 to 5 consecutive days (N = 5 rats per group). Rats swam in basins 47 cm in diameter and 60 cm deep filled with water at 33 to 35ºC. After the training period a significant increase (P &lt; 0.05) was observed in the heart weight normalized to body weight by about 22 and 35% in the groups that trained for 96 and 120 h, respectively. Blood lactate levels were significantly increased (P &lt; 0.05) in all groups after all training...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Myosin heavy chain expression; Training; Real time PCR; Rat myocardium; Muscle hypertrophy.
Ano: 2006
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000500009

Etude des effets de polluants sur les paramètres hémocytaires de l'huître creuse, Crassostrea gigas - Interactions entre environnement, mécanismes de défense et maladies infectieuses
Provedor de dados: 5
Autores: Gagnaire, Beatrice.
Shellfish industry is mostly realized in estuary zones, which are subjected to pollutions due to anthropic activities. The harmful effects of pollutants on animals inhabiting these estuarine zones are poorly known. Among these animals, Pacific oyster, Crassostrea gigas, may represent a model because they are sedentary and they filter water intensively. Among all physiological functions possibly disturbed by pollutants, defence mechanisms are poorly studied in bivalves. Moreover, animals presenting impaired defence mechanisms may be more sensitive to infectious diseases. In this context, effects of pollutants on hemocyte functions and on sensitiveness to diseases were tested in C. gigas. After adjusting the protocols for the monitoring of hemocyte...
Tipo: Text
Palavras-chave: Real time PCR; Flow cytometry; Pesticides; Pollutants; Phagocytosis; Hemocytes; Crassostrea gigas; Pacific oyster; Pathology; Immunotoxicology; PCR en temps réel; Cytométrie de flux; Pesticides; Polluants; Phagocytose; Hémocytes; Crassostrea gigas; Huître creuse; Pathologie; Immunotoxicologie.
Ano: 2005
URL: http://archimer.ifremer.fr/doc/2005/these-730.pdf

Expression analysis for genes involved in arachidonic acid biosynthesis in Mortierella alpina CBS 754.68
Provedor de dados: 58
Autores: Samadlouie,Hamid-Reza; Hamidi-Esfahani,Zohreh; Alavi,Seyed-Mehdi; Varastegani,Boshra.
The time courses for production of fungal biomass, lipid, phenolic and arachidonic acid (ARA) as well as expression of the genes involved in biosynthesis of ARA and lipid were examined in Mortierella alpina CBS 754.68. A significant increase in the arachidonic acid content in lipids that coincided with reduced levels of lipid was obtained. Reduced gene expression occurred presumably due to the steady reduction of carbon and nitrogen resources. However, these energy resources were inefficiently compensated by the breakdown of the accumulated lipids that in turn, induced up-regulated expression of the candidate genes. The results further indicated that the expression of the GLELO encoding gene is a rate-limiting step in the biosynthesis of ARA in the early...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Mortierella alpina; Arachidonic acid; Lipid; Gene expression; Real time PCR.
Ano: 2014
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200010

First description of French V. tubiashii strains pathogenic to mollusk: I. Characterization of isolates and detection during mortality events
Provedor de dados: 5
Autores: Travers, Marie-agnes; Mersni Achour, Rachida; Haffner, Philippe; Tourbiez, Delphine; Cassone, Anne-laure; Morga, Benjamin; Doghri, Ibtissem; Garcia, Celine; Renault, Tristan; Fruitier-arnaudin, Ingrid; Saulnier, Denis.
Nine dominant bacterial isolates were obtained from different batches of Crassostrea gigas spat experiencing high mortality rates in a French experimental hatchery/nursery in 2007. Using phenotypic analysis combined with multilocus sequence analysis, the isolates were shown to be genetically close to the Vibrio tubiashii type strain. Based on (1) analyses of the recA gene sequences; (2) the results of DNA–DNA hybridization assays between 07/118 T2 (LMG 27884 = CECT 8426), which is a representative strain, and the V. tubiashii type strain (69%); and (3) phenotypic traits, the bacteria were classified in a group close to American V. tubiashii strain. Its virulence (70% of mortalities) and the toxicity of the extracellular products of 07/118 T2 was...
Tipo: Text
Palavras-chave: Crassostrea gigas Pathogenicity Extracellular products Polyphasic approach; Real time PCR.
Ano: 2014
URL: http://archimer.ifremer.fr/doc/00189/30028/28743.pdf

Identification of a splicing coactivator gene that affects the production of ochratoxin a in Aspergillus carbonarius
Provedor de dados: 52
Autores: Lunardi,Lígia Uno; Guembarovski,Roberta Losi; Hanai,Luiz Ricardo; Cristiano,Valderi; Vieira,Maria Lucia Carneiro; Sartori,Daniele; Fungaro,Maria Helena Pelegrinelli.
Ochratoxin A is a mycotoxin produced by some fungi species. Among them, Aspergillus carbonarius is considered a powerful producer. Genes involved in the ochratoxin A biosynthesis pathway have been identified in some producer species. However, there are few studies that purpose to identify these genes in A. carbonarius. The use of insertion mutants to identify genes associated with certain properties has been increased in the literature. In this work, the region of T-DNA integration was investigated in one A. carbonarius ochratoxin-defective mutant previously obtained by Agrobacterium tumefaciens-mediated transformation, in order to find an association between interrupted gene and the biosynthesis of ochratoxin A. The integration occurred in a gene that...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Ochratoxigenic fungi; Mycotoxin; Real time PCR; Splicing coactivator protein.
Ano: 2009
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700018

Identification of genes from flat oyster Ostrea edulis as suitable housekeeping genes for quantitative real time PCR
Provedor de dados: 5
Autores: Morga, Benjamin; Arzul, Isabelle; Faury, Nicole; Renault, Tristan.
Bonamia ostreae is an intrahaemocytic protozoan affecting Ostrea edulis. The parasite multiplies within haemocytes without being degraded and involves changes in cellular activities. Studies aiming at better understanding host response to a pathogen at the transcriptome levels are frequently based on the use of real time PCR assays, which require some reference genes. However, very few sequence data is available for O. edulis in public databases.  Subtracted cDNA libraries were constructed from the O. edulis haemocytes in order to identify genes involved in host reactions against the parasite and quantitative real time PCR assays were developed to study expression of these genes. In this context, identification of reference genes and study of their...
Tipo: Text
Palavras-chave: Real time PCR; Housekeeping genes; Haemocytes; Ostrea edulis; Bonamia ostreae.
Ano: 2010
URL: http://archimer.ifremer.fr/doc/00018/12880/9829.pdf

Molecular detection and quantification of the protozoan Bonamia ostreae in the flat oyster, Ostrea edulis
Provedor de dados: 5
Autores: Robert, Maeva; Garcia, Celine; Chollet, Bruno; Lopez-flores, Inmaculada; Ferrand, Sylvie; Francois, Cyrille; Joly, Jean-pierre; Arzul, Isabelle.
Bonamia ostreae is an intracellular protozoan which is recognized as a cause of mortality in European populations of flat oysters (Ostrea edulis). Based on the recent characterization of actin genes of B. ostreae, specific primers were designed for real-time PCR using SYBR® Green chemistry. Specificity was demonstrated by the unique melting temperature peak observed in positive samples and by the lack of amplification in samples of oysters infected by closely related parasites, including Bonamia exitiosa. A calibration curve using a cloned template was defined to estimate copy number. The assay had a 6 log- dynamic range, mean inter- and intra-assay variation coefficients of &lt;1% and a minimum detection limit of 50 gene copies per reaction. Using...
Tipo: Text
Palavras-chave: Heart imprints; Ostrea edulis; Quantification; Detection; Real time PCR; Bonamia ostreae.
Ano: 2009
URL: http://archimer.ifremer.fr/doc/2009/publication-6934.pdf

Morphology and expression of genes related to skeletal muscle growth in juveniles of pirarucu (Arapaima gigas, Arapaimatidae, Teleostei) - doi: 10.4025/actascianimsci.v35i3.18219
Provedor de dados: 2
Autores: Carani, Fernanda Regina; Universidade Estadual Paulista (UNESP); Duran, Bruno Oliveira da Silva; Universidade Estadual Paulista (UNESP); Paula, Tassiana Gutierrez de; Universidade Estadual Paulista (UNESP); Piedade, Warlen Pereira; Universidade Estadual Paulista (UNESP); Dal-Pai, Maeli; Universidade Estadual Paulista (UNESP).
Skeletal muscle growth in the pirarucu (Arapaima gigas) is highly interesting to fish farmers because it provides information about how the mechanism in muscle mass increase, characteristic of the species, is regulated. Pirarucu has specific muscle growth that highlights the species’s significance and commercial value. Current research evaluates the morphology and the growth-related gene expression in the red and white skeletal muscles of the pirarucu. Muscle samples were collected from the lateral anterior region and frozen in liquid nitrogen. Histological sections were performed and stained by HE for morphological analysis. Red and white muscle samples were used to determine MyoD, myogenin, and myostatin genes expression by Real-time Polymerase Chain...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: 5.06.03.00-0 muscle fibre morphology; Myogenic regulatory factors; Myostatin; Skeletal muscle growth; Arapaima gigas; Real time PCR.
Ano: 2013
URL: http://periodicos.uem.br/ojs/index.php/ActaSciAnimSci/article/view/18219

Mycobacterium avium subsp. Paratuberculosis and the expression of selected virulence and pathogenesis genes in response to 6°c, 65°c and ph 2.0
Provedor de dados: 58
Autores: Pribylova,Radka; Kralik,Petr; Donnelly,Neysan; Matiasovic,Jan; Pavlik,Ivo.
The aim of this work was to study the expression of selected Mycobacterium avium subsp. paratuberculosis (MAP) genes connected with MAP virulence, adhesion and stress response. The temperature of 6°C and 65°C were chosen with regard to the food industry, storage conditions (refrigerator) and low-temperature pasteurization. A pH of 2.0, using lactic acid, was selected to mimic the natural environment of the stomach. Expression of selected genes was studied using real time reverse transcription PCR on three different MAP isolates. MAP isolates were chosen according to the number of their preceding cultivations. While isolates 8672 and 8819 were previously cultivated only once, MAP isolate 12146 went through four passages. Different expression profiles were...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: MAP; Johne's disease; Stress; Treatment; Real time PCR.
Ano: 2011
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000200049

Ostreid herpes virus 1 infection in families of the Pacific oyster, Crassostrea gigas, during a summer mortality outbreak: Differences in viral DNA detection and quantification using real-time PCR
Provedor de dados: 5
Autores: Sauvage, Christopher; Pepin, Jean-francois; Lapegue, Sylvie; Boudry, Pierre; Renault, Tristan.
Ostreid herpes virus 1 (OsHV-1) infections, notably reported in Europe and the USA, are closely associated with significant mortalities of the Pacific oyster, Crassostrea gigas, especially during its early stages of life. In summer 2006, we monitored mortality by strict daily verification of three full-sib families of oysters reared under common conditions. We quantified OsHV-1 using real-time PCR in dead and living individuals during and after a mortality event. Mortality events were severe and brief, but significantly different between tested families (cumulative mortality ranging from 1.2 to 49%). Real-time PCR assays revealed different viral DNA loads in dead individuals from different families (P &lt; 0.001). Moreover, the mean level of infection...
Tipo: Text
Palavras-chave: Viral DNA quantification; Real time PCR; OsHV 1; Mortality; Crassostrea gigas.
Ano: 2009
URL: http://archimer.ifremer.fr/doc/2009/publication-6265.pdf

Quantification of Vibrio penaeicida, the etiological agent of Syndrome 93 in New Caledonian shrimp, by real-time PCR using SYBR Green I chemistry
Provedor de dados: 5
Autores: Goarant, Cyrille; Merien, F.
Shrimp farming is a small but growing industry in New Caledonia. Since 1993, "Syndrome 93" has been affecting New Caledonian shrimp farming industry every cold season, causing severe epizootic mortalities in grow-out ponds and significant losses. Highly pathogenic strains of Vibrio penaeicida are considered the etiological agent of the disease in Litopenaeus stylirostris. On one hand, studies demonstrated that healthy shrimp may carry V penaeicida for weeks with a high overall prevalence, regardless of any seasonal pattern or temperature conditions. On the other hand, larvae are free of V penaeicida and are also resistant to experimental infection. V penaeicida is frequently detected in incoming water pumped from the bays, which was shown, by a molecular...
Tipo: Text
Palavras-chave: Vibriosis; Vibrio; Real time PCR; Quantification; Mariculture; Extraction techniques.
Ano: 2006
URL: http://archimer.ifremer.fr/doc/2006/publication-1903.pdf

Quantitative differential expression of alpha and beta ryanodine receptor genes in PSE (Pale, Soft, Exudative) meat from two chicken lines: broiler and layer
Provedor de dados: 52
Autores: Oda,Sandra Helena Inoue; Nepomuceno,Alexandre Lima; Ledur,Mônica Corrêa; Oliveira,Maria Cristina Neves de; Marin,Silvana Regina Rockenbach; Ida,Elza Iouko; Shimokomaki,Massami.
Total RNA isolated from Pectoralis major muscle from PSE (L*24h&gt;53.0, pH&lt;5.8) and non-PSE (44&lt;"L*24h&gt;"53) meats of two phenotypically distinct chicken lines, broiler and layer, was used to investigate the α-ryr and β-ryr gene expression by real-time RT-PCR approach. Mean relative quantification (RQ) values were lower (p&lt;0.05) for β-ryr in PSE chickens from both lines when compared to non-PSE chickens, while there was no difference (p&gt;0.05) in α-ryr gene expression regardless of line studied. The β-ryr RQ results suggested that in PSE samples an alteration might occur in the regular ratio (1:1) of α-RyR/β-RyR normally found in avian muscles. These results provided the first evidence of PSE meat occurrence as a result of the differential...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Malignant hyperthermia; Poultry; Gene expression; Real time PCR.
Ano: 2009
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000600024

Rapid and sensitive detection of ostreid herpesvirus 1 in oyster samples by real-time PCR
Provedor de dados: 5
Autores: Pepin, Jean-francois; Riou, Antoine; Renault, Tristan.
Herpes and herpes-like virus infections have been reported in various marine mollusc species associated with high mortality rates. Following the characterisation and genome sequencing of ostreid herpesvirus 1 (OsHV-1), specific diagnostic tools have been developed based on conventional PCR techniques or in situ hybridisation. We have now developed a real-time PCR assay for rapid, sensitive and quantitative detection of OsHV-1, and compared it with a conventional PCR technique described previously. The new assay utilised SYBR® Green chemistry with specific primers C9/C10 targeting the C region. The melt curve analysis of OsHV-1 DNA or DNA extracted from infected material showed only one melting temperature peak (75.75 ± 0.1 °C). The assay had a detection...
Tipo: Text
Palavras-chave: SYBR® Green; Real time PCR; Crassostrea gigas; Ostreid herpesvirus 1; Herpesvirus.
Ano: 2008
URL: http://archimer.ifremer.fr/doc/2008/publication-3951.pdf

Real-time PCR assay for rapid detection and quantification of Vibrio aestuarianus in oyster and seawater: A useful tool for epidemiologic studies
Provedor de dados: 5
Autores: Saulnier, Denis; De Decker, Sophie; Haffner, Philippe.
Because Vibrio aestuarianus is known to cause serious infections in Pacific oyster Crassostrea gigas, a real-time PCR assay was developed targeting the dnaJ gene of this bacterium. Only V. aestuarianus strains isolated from C. gigas mortality events in different geographic areas and the reference strain tested positive, whereas no amplification products was obtained with type strains belonging to 23 other species of Vibrio. Sensitivity and reproducibility of the method were assessed using either seawater or oyster homogenate samples spiked with one V aestuarianus strain. All these samples were stored at -20 degrees C in order to mimic retrospective or grouped natural sample analysis without quantification bias due to prolonged freezing. Analysis of...
Tipo: Text
Palavras-chave: V. aestuarianus; Vibrio; Taqman; Real time PCR; Pathogen; Oyster; Crassostrea gigas.
Ano: 2009
URL: http://archimer.ifremer.fr/doc/2009/publication-6447.pdf

Sequence polymorphism-based identification and quantification of Vibrio nigripulchritudo at the species and subspecies level targeting an emerging pathogen for cultured shrimp in New Caledonia
Provedor de dados: 5
Autores: Goarant, Cyrille; Reynaud, Yann; Ansquer, Dominique; De Decker, Sophie; Merien, Fabrice.
In a previous study, we demonstrated the existence of an emerging cluster of Vibrio nigripulchritudo that proved to be associated with shrimp mortality events in New Caledonia. Using sequence polymorphisms evidenced in this previous MultiLocus Sequence Typing study, we developed two new quantitative PCR assays permitting the detection and quantification of V. nigripulchritudo at the genospecies level using SYBR Green I chemistry and at the emerging cluster level using Fluorescence Resonance Energy Transfer technology with hybridization probes.   The use of this molecular diagnostic tool evidenced the colonization of the shrimp pond ecosystem by the pathogenic cluster at least at the onset of the disease. This new tool will allow better investigation of the...
Tipo: Text
Palavras-chave: Shrimp; Cluster specific; Vibrio; Pathogen; Mariculture; Real time PCR.
Ano: 2007
URL: http://archimer.ifremer.fr/doc/2007/publication-2729.pdf

The effect of environmental salinity on the proteome of the sea bass (Dicentrarchus labrax L.)
Provedor de dados: 5
Autores: Ky, Chin Long; De Lorgeril, Julien; Hirtz, C; Sommerer, N; Rossignol, M; Bonhomme, F.
The European sea bass, Dicentrarchus labrax L., tolerates a range of salinities from freshwater to hyper-saline. To study differences in protein expression, fish were reared in both freshwater and seawater. After 3-month acclimation, gill and intestine epithelia were collected and the soluble protein extracted. In all, 362 spots were differentially expressed in the gills and intestines of fishes reared in seawater compared to those from freshwater. Fifty differential protein spots were excised from a colloidal Coomassie-stained gel. Nine separate protein spots were identified unambiguously by mass spectrometry and database searching. Among the six proteins over-expressed in gill cells in seawater, five were cytoskeleton proteins and one was the aromatase...
Tipo: Text
Palavras-chave: Two dimensional gel electrophoresis; Sea bass; Salinity; Real time PCR; MALDI TOF mass spectrometry.
Ano: 2007
URL: http://archimer.ifremer.fr/doc/2007/publication-3952.pdf

Transcript levels of ten caste-related genes in adult diploid males of Melipona quadrifasciata (Hymenoptera, Apidae): a comparison with haploid males, queens and workers
Provedor de dados: 74
Autores: Borges,Andreia A.; Humann,Fernanda C.; Campos,Lucio A. Oliveira; Tavares,Mara G.; Hartfelder,Klaus.
In Hymenoptera, homozygosity at the sex locus results in the production of diploid males. In social species, these pose a double burden by having low fitness and drawing resources normally spent for increasing the work force of a colony. Yet, diploid males are of academic interest as they can elucidate effects of ploidy (normal males are haploid, whereas the female castes, the queens and workers, are diploid) on morphology and life history. Herein we investigated expression levels of ten caste-related genes in the stingless bee Melipona quadrifasciata, comparing newly emerged and 5-day-old diploid males with haploid males, queens and workers. In diploid males, transcript levels for dunce and paramyosin were increased during the first five days of adult...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Stingless bee; Real time PCR; Caste; Diploid male; Differential gene expression.
Ano: 2011
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000400025

Transcriptomic study of 39 ostreid herpesvirus 1 genes during an experimental infection
Provedor de dados: 5
Autores: Segarra, Amelie; Faury, Nicole; Pepin, Jean-francois; Renault, Tristan.
Massive mortality outbreaks have been reported in France since 2008 among Pacific oysters, Crassostrea gigas, with the detection of a particular OsHV-1 variant called μVar. Virus infection can be induced in healthy spat in experimental conditions allowing to better understand the disease process, including viral gene expression.  Although gene expression of other herpesviruses has been widely studied, we provide the first study following viral gene expression of OsHV-1 over time. In this context, an in vivo transcriptomic study targeting 39 OsHV-1 genes was carried out during an experimental infection of Pacific oyster spat. For the first time, several OsHV-1 mRNAs were detected by real-time PCR at 0 h, 2 h, 4 h, 18 h, 26 h and 42 h post injection. Several...
Tipo: Text
Palavras-chave: Crassostrea gigas; OsHV-1; Viral gene expression; Inhibitors of apoptosis; Real time PCR; Elongation factor.
Ano: 2014
URL: http://archimer.ifremer.fr/doc/00184/29549/27875.pdf

Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis
Provedor de dados: 58
Autores: Sales,Mariana L.; Fonseca Júnior,Antônio Augusto; Orzil,Lívia; Alencar,Andrea Padilha; Silva,Marcio Roberto; Issa,Marina Azevedo; Soares Filho,Paulo Martins; Lage,Andrey Pereira; Heinemann,Marcos Bryan.
Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Mycobacterium tuberculosis; Diagnosis; Real time PCR.
Ano: 2014
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400029