Consulta: subjectFacets:"Quantitative PCR"
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Data/hora: 07/06/2024 10:56:57

A quantitative PCR approach for determining the ribosomal DNA copy number in the genome of Agave tequila Weber
Provedor de dados: 69
Autores: Rubio-Piña,Jorge; Quiroz-Moreno,Adriana; Sánchez-Teyer,L. Felipe.
Background: Agave tequilana has a great economic importance in Mexico in order to produce alcoholic beverages and bioenergy. However, in this species the structure and organization of the rDNAs in the genome are limited, and it represents an obstacle both in their genetic research and improvement as well. rDNA copy number variations per eukaryotic genome have been considered as a source of genetic rearrangements. In this study, the copy number of 18S and 5S rDNAs in the A. tequilana genome was estimated, and an absolute quantitative qPCR assay and genome size was used. In addition, an association between the rDNAs copy number and physical mapping was performed to confirm our results. Results: The analysis were successfully applied to determine copy number...
Tipo: Journal article
Palavras-chave: Agave tequilana; Copy number; Fluorescent in situ hybridization; Ribosomal DNA; Quantitative PCR.
Ano: 2016
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000400002

Simultaneous quantitative assessment of circulating cell-free mitochondrial and nuclear DNA by multiplex real-time PCR
Provedor de dados: 74
Autores: Xia,Peng; Radpour,Ramin; Zachariah,Rebecca; Fan,Alex Xiu Cheng; Kohler,Corina; Hahn,Sinuhe; Holzgreve,Wolfgang; Zhong,Xiao Yan.
Quantification of circulating nucleic acids in plasma and serum could be used as a non-invasive diagnostic tool for monitoring a wide variety of diseases and conditions. We describe here a rapid, simple and accurate multiplex real-time PCR method for direct synchronized analysis of circulating cell-free (ccf) mitochondrial (mtDNA) and nuclear (nDNA) DNA in plasma and serum samples. The method is based on one-step multiplex real-time PCR using a FAM-labeled MGB probe and primers to amplify the mtDNA sequence of the ATP 8 gene, and a VIC-labeled MGB probe and primers to amplify the nDNA sequence of the glycerinaldehyde-3-phosphate-dehydrogenase (GAPDH) gene, in plasma and serum samples simultaneously. The efficiencies of the multiplex assays were measured in...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Circulating cell-free DNA; Mitochondrial DNA; Nuclear DNA; Real-time PCR; Quantitative PCR.
Ano: 2009
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572009000100003

Molecular approach indicates consumption of jellyfish by commercially important fish species in a coastal Mediterranean lagoon
Provedor de dados: 5
Autores: Marques, Raquel; Darnaude, Audrey M.; Crochemore, Sandrine; Bouvier, Corinne; Bonnet, Delphine.
Until recently, jellyfish have been ignored as an important source of food, due to their low nutritional value. Here, quantitative PCR was used to detect and quantify the DNA of the jellyfish Aurelia coerulea in the gut contents of commercially important fish species from the Thau Lagoon. Individuals from five fish species were collected during two different periods: the bloom period, when the pelagic stages of A. coerulea are abundant, and the post-bloom period, when only the benthic stage – polyps – is present in the lagoon. The DNA of A. coerulea was detected in the guts of 41.9% of the fish analysed, belonging to four different species. The eel Anguilla anguilla and the seabream Sparus aurata were important jellyfish consumers during the bloom and...
Tipo: Text
Palavras-chave: Predation; Aurelia coerulea; Eel; Seabream; Polyps; Medusae; Quantitative PCR; Gut content; Thau lagoon.
Ano: 2019
URL: https://archimer.ifremer.fr/doc/00513/62469/66986.pdf

Comparative assessment of human and farm animal faecal microbiota using real-time quantitative PCR
Provedor de dados: 5
Autores: Furet, Jean-pierre; Firmesse, Olivier; Gourmelon, Michele; Bridonneau, Chantal; Tap, Julien; Mondot, Stanislas; Dore, Joel; Corthier, Gerard.
Pollution of the environment by human and animal faecal pollution affects the safety of shellfish, drinking water and recreational beaches. To pinpoint the origin of contaminations, it is essential to define the differences between human microbiota and that of farm animals. A strategy based on real-time quantitative PCR (qPCR) assays was therefore developed and applied to compare the composition of intestinal microbiota of these two groups. Primers were designed to quantify the 16S rRNA gene from dominant and subdominant bacterial groups. TaqMan((R)) probes were defined for the qPCR technique used for dominant microbiota. Human faecal microbiota was compared with that of farm animals using faecal samples collected from rabbits, goats, horses, pigs, sheep...
Tipo: Text
Palavras-chave: Quantitative PCR; Faecal microbiota; Human; Farm animals.
Ano: 2009
URL: http://archimer.ifremer.fr/doc/00000/11071/11297.pdf

A novel duplex qPCR assay for stepwise detection of multiple Perkinsea protistan infections of amphibian tissues
Provedor de dados: 5
Autores: Smilansky, Vanessa; Chambouvet, Aurelie; Reeves, Mari; Richards, Thomas A.; Milner, David S.
Alveolate protists within the phylum Perkinsea have been found to infect amphibians across a broad taxonomic and geographic range. Phylogenetic analysis has suggested the existence of two clades of amphibian Perkinsea: a putatively non-pathogenic clade linked to asymptomatic infections of tadpoles in Africa, Europe and South America, and a putatively pathogenic clade linked to disease and mass mortality events of tadpoles in North America. Here, we describe the development of a duplex TaqMan qPCR assay to detect and discriminate between rDNA sequences from both clades of Perkinsea in amphibian tissues. The assay uses a single primer pair to target an 18S small subunit (SSU) ribosomal RNA (rRNA) gene region shared between the two clades, and two...
Tipo: Text
Palavras-chave: Frog disease; Alveolate parasites; Quantitative PCR; Perkinsea; NAG01.
Ano: 2021
URL: https://archimer.ifremer.fr/doc/00685/79758/82558.pdf

Assessment of DNA extraction methods from GMO analysis for grain monitoring in Mexico: Part II: quantification by real-time PCR
Provedor de dados: 115
Autores: Acatzi,Abraham; Galvez,Amanda; Plasencia,Javier; Quirasco,Maricarmen.
Because of their specificity and high throughput, real-time PCR-based methods are suitable for the monitoring of genetically modified (GM) organisms at field level or in the grain trade, in order to comply with federal regulations on biosafety. In the first part of this study, DNA extracted from different maize (Zea mays L.) tissues using several commercial purification protocols available in Mexico were evaluated in terms of DNA quality as substrates for end-point PCR. In this second part, DNA preparations obtained from grain, by means of the same commercial protocols, were tested in quantitative PCR (qPCR), using TaqMan and SYBR Green protocols. Linear dynamic range, amplification efficiency and method accuracy were assessed using recommended qPCR...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Quantitative PCR; PCR inhibition; GM maize; Genetically modified organism (GMO); DNA quality.
Ano: 2014
URL: http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S1405-31952014000100003

Population Changes of Eubacteria, Sulfate-Reducing Bacteria and Methanogenic Archaea in an Anaerobic Reactor Processing Ethanol Distillery Vinasse
Provedor de dados: 52
Autores: Brown,Adis Ivonne Terry; Pozzi,Eloisa; Damianovic,Marcia Helena Rissato Zamariolli; Briones,Homero Enrique Urrutia; Ortiz,Leslie Ester Abarzúa; Pires,Eduardo Cleto.
Abstract The microbiological characterization by molecular techniques (DGGE and quantitative PCR) of Archaea, Bacteria domain and sulfate-reducing bacteria (SRB) from a laboratory scale Up-flow Anaerobic Sludge Blanket (UASB) processing sugar cane vinasse was performed during the operational phase with increasing organic loads. The organic load removal efficiency was between 97% and 75% for volumetric organic loads (VOL) in the range of 0.6 to 15.4 kgCOD.m-3.d−1 and for higher VOL (until 27.0 kgCOD.m-3 .d−1) the removal efficiency decreased to 48%. Archaea represented the majority of the estimated population (107 copies of 16S RNA ribosomal gene. mL-1) followed by bacteria (106 copies of 16S RNA ribosomal gene. mL-1) and sulfate-reducing bacteria (SRB)...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: UASB reactor; Microbial characterization; Methanogenesis; Sulfidogenesis; DGGE; Quantitative PCR.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100203

Fecal indicators and bacterial pathogens in bottled water from Dhaka, Bangladesh
Provedor de dados: 58
Autores: Ahmed,W.; Yusuf,R.; Hasan,I.; Ashraf,W.; Goonetilleke,A.; Toze,S.; Gardner,T.
Forty-six bottled water samples representing 16 brands from Dhaka, Bangladesh were tested for the numbers of total coliforms, fecal indicator bacteria (i.e., thermotolerant Escherichia coli and Enterococcus spp.) and potential bacterial pathogens (i.e., Aeromonas hydrophil, Pseudomonas aeruginos, Salmonella spp., and Shigella spp.). Among the 16 brands tested, 14 (86%), ten (63%) and seven (44%) were positive for total coliforms, E. coil and Enterococcus spp., respectively. Additionally, a further nine (56%), eight (50%), six (37%), and four (25%) brands were PCR positive for A. hydrophila lip, P. aeruginosa ETA, Salmonella spp. invA, and Shigella spp. ipaH genes, respectively. The numbers of bacterial pathogens in bottled water samples ranged from 28 ± 12...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Bottled water; Fecal indicator bacteria; Quantitative PCR; Bacterial pathogens; Public health risk.
Ano: 2013
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822013000100013

Malignant Catarrhal Fever in Brazilian cattle presenting with neurological syndrome
Provedor de dados: 58
Autores: Martins,Maira de S.N.; Castro,Alessandra M.M.G. de; Lima,Michele dos S.; Pinto,Vivian da S.C.; Silva,Thaís G. da; Fava,Claudia Del; Depes,Claudio Regis; Okuda,Liria H.; Pituco,Edviges M.
Abstract Malignant Catarrhal Fever (MCF) was investigated in the central nervous system of cattle with neurological syndrome. Two-hundred-ninety samples were analyzed by histology, and molecular methods to detect ovine herpesvirus type 2 (OvHV-2) were optimized and validated. The qualitative polymerase chain reaction (qualitative PCR) analytical sensitivity was 101 DNA copies/µL and found 4.8% (14/290) positive for OvHV-2. The quantitative polymerase chain reaction (qPCR) analytical sensitivity was 100 DNA copy/µL and 5.9% (17/290) positivity, with 47.1% (8/17) of the positive samples presenting histological evidence of non-purulent meningo-encephalitis. The qualitative PCR products (422 bp of the ORF75 region) were sequenced and submitted to phylogenetic...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Ovine herpesvirus type 2 (OvHV-2); Histopathology; Qualitative PCR; Quantitative PCR; Phylogenetic analysis.
Ano: 2017
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000200366

Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis
Provedor de dados: 58
Autores: Chen,Chun; Xie,Tingna; Ye,Sudan; Jensen,Annette Bruun; Eilenberg,J ørgen.
Abstract The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Pandora neoaphidis; Reference genes; Quantitative PCR.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000100259

[RETRACTED ARTICLE] Bovine Vaccinia in dairy cattle and suspicion of vesicular disease on milkers in Brazil
Provedor de dados: 65
Autores: Silva,Thaís Garcia da; Lima,Michele dos Santos; Castro,Alessandra Marnie Martins Gomes de; Martins,Maira de Souza Nunes; Castiglioni,Vivian Cardoso; Fava,Claudia Del; Okuda,Liria Hiromi; Pituco,Edviges Maristela.
ABSTRACT: Bovine vaccinia (BV) is a vesicular disease induced by the Vaccinia virus (VACV) that affects milk production and is an occupational zoonosis. This research had the following objectives: (i) detection of VACV by qPCR in cattle with clinical suspicion of vesicular disease; (ii) symptoms characterization in animals and milkers with clinical suspicion of the disease and virus detection in humans; and (iii) identification of risk factors for infections of VACV in herds from several Brazilian states. A total of 471 bovine epithelial samples from dairy farms, in 15 Brazilian states, were evaluated between 2007 and 2012. The samples were tested by quantitative PCR (qPCR) using SYBR Green® reagents, validated with a lower limit of detection of...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Orthopoxvirus; Poxviridae; Quantitative PCR; Risk factor; VACV; Zoonosis.
Ano: 2018
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782018000600455

Bovine Vaccinia in dairy cattle and suspicion of vesicular disease on milkers in Brazil
Provedor de dados: 65
Autores: Silva,Thaís Garcia da; Lima,Michele dos Santos; Castro,Alessandra Marnie Martins Gomes de; Martins,Maira de Souza Nunes; Castiglioni,Vivian Cardoso; Fava,Claudia Del; Okuda,Liria Hiromi; Pituco,Edviges Maristela.
ABSTRACT: Bovine vaccinia (BV) is a vesicular disease induced by the Vaccinia virus (VACV) that affects milk production and is an occupational zoonosis. This research had the following objectives: (i) detection of VACV by qPCR in cattle with clinical suspicion of vesicular disease; (ii) symptoms characterization in animals and milkers with clinical suspicion of the disease and virus detection in humans; and (iii) identification of risk factors for infections of VACV in herds from several Brazilian states. A total of 471 bovine epithelial samples from dairy farms, in 15 Brazilian states, were evaluated between 2007 and 2012. The samples were tested by quantitative PCR (qPCR) using SYBR Green® reagents, validated with a lower limit of detection of 100...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Orthopoxvirus; Poxviridae; Quantitative PCR; Risk factor; VACV; Zoonosis.
Ano: 2018
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782018000500451