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| Registros recuperados: 28 | |
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| 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 |
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| 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 |
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| 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 |
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| 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 |
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| 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 |
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| 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 |
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| 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 |
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| 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 |
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| 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 <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 |
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| 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 |
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| 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 < 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 |
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| 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 |
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| Registros recuperados: 28 | |
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