Search results for "PCR"
showing 10 items of 438 documents
Becton Dickinson Directigen EZ Flu A+B assay in the diagnosis of pandemic influenza A H1N1 2009 virus infection in adult patients
2011
Structural and functional diversity of Bacillus thuringiensis toxins on animal, plant and microbial cells
2018
Debido a los efectos nocivos de los insecticidas químicos en los ecosistemas y sobre los organismos beneficiosos, la implementación de agentes de control biológico ha recibido considerable atención. Entre los agentes de control biológico, se ha dedicado gran interés a los entomopatógenos, debido a su potencial para controlar plagas agrícolas y domésticas, vectores de enfermedades humanas y animales, sin introducir material no degradable en el medio ambiente. Bacillus thuringiensis (Bt) es el agente de control microbiano comercial más utilizado en la actualidad, con efectividad contra diferentes especies de insectos plaga. Las cepas de Bt producen cristales paraesporales proteináceos durante…
Validation of a set of reference genes to study response to herbicide stress in grasses
2012
Abstract Background Non-target-site based resistance to herbicides is a major threat to the chemical control of agronomically noxious weeds. This adaptive trait is endowed by differences in the expression of a number of genes in plants that are resistant or sensitive to herbicides. Quantification of the expression of such genes requires normalising qPCR data using reference genes with stable expression in the system studied as internal standards. The aim of this study was to validate reference genes in Alopecurus myosuroides, a grass (Poaceae) weed of economic and agronomic importance with no genomic resources. Results The stability of 11 candidate reference genes was assessed in plants res…
DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen
2020
Droplet microfluidics is a technology that enables the production and manipulation of small volumes. In biosciences, the most popular application of this technology is Droplet Digital™ PCR (ddPCR™), where parallel nanoliter-scale PCR assays are used to provide a high sensitivity and specificity for DNA detection. However, the recovery of PCR products for downstream applications such as sequencing can be challenging due to the droplets' stability. Here we compared five methods for disrupting the droplets to recover DNA. We found that rapid freezing in liquid nitrogen results in a clear phase separation and recovery of up to 70% of the DNA content. Liquid nitrogen freezing can thus offer a si…
Development of molecular approaches for direct detection and quantification of wine-related microorganisms
2018
En esta tesis doctoral se han desarrollado procedimientos para la detección y cuantificación de levaduras y bacterias acéticas totales y, en concreto, de las especies más importantes en la vinificación como Saccharomyces cerevisiae, Brettanomyces bruxellensis, Zygosaccharomyces bailii, L. plantarum, y O. oeni. Las estrategias que se han utilizado para el desarrollo de nuevos métodos se centran fundamentalmente en la supresión del paso de extracción previa de ADN y de los procedimientos de eliminación de inhibidores de la reacción de PCR presentes en las muestras. Para llevar a cabo este objetivo general, se establecieron los siguientes objetivos específicos; 1, desarrollo de un método de qP…
Clinical significance of the polymerase chain reaction (PCR) assay in chronic HBV carriers
1992
PCR was evaluated as a clinical tool for use in accurate identification of the specific etiologic agent in chronic HBV carriers. The method was found to be valuable in diagnosis and for monitoring therapy, as well as for elucidation of genotypic variants of HBV in chronic HBV cases. By this means an HBV defective variant with alterations in the preSl/preS2 sequence was detected and is consequently described here.
A One-Step PCR-Based Assay to Evaluate the Efficiency and Precision of Genomic DNA-Editing Tools
2017
Despite rapid progress, many problems and limitations persist and limit the applicability of gene-editing techniques. Making use of meganucleases, TALENs, or CRISPR/Cas9-based tools requires an initial step of pre-screening to determine the efficiency and specificity of the designed tools. This step remains time consuming and material consuming. Here we propose a simple, cheap, reliable, time-saving, and highly sensitive method to evaluate a given gene-editing tool based on its capacity to induce chromosomal translocations when combined with a reference engineered nuclease. In the proposed technique, designated engineered nuclease-induced translocations (ENIT), a plasmid coding for the DNA-…
Análisis molecular de la valoración del intervalo de tiempo óptimo entre la administración de acetato de triptorelina y la punción folicular en los t…
2023
Los fármacos análogos de la GnRH y la hCG han demostrado ser igualmente eficaces en la activación de la cascada de intermediarios de la maduración final ovocitaria que ocurre tras el estímulo ovulatorio. Sin embargo, su mecanismo de acción y por tanto los perfiles hormonales hallados tras el estímulo de los dos fármacos son muy diferentes. El tiempo más adecuado para programar la recolección ovocitaria tras la administración de acetato de triptorelina podría no ser 36 horas como ampliamente se ha pautado en los tratamientos de FIV con hCG. Este intervalo de tiempo es sumamente importante para obtener la mayor proporción posible de ovocitos MII porque procesos como el inicio de la luteinizac…
Implementation of Sample Pooling Procedure Using a Rapid SARS-CoV-2 Diagnostic Real-Time PCR Test Performed Prior to Hospital Admission of People wit…
2021
Reliability, accuracy, and timeliness of diagnostic testing for SARS-CoV-2 infection have allowed adequate public health management of the disease, thus notably helping the timely mapping of viral spread within the community. Furthermore, the most vulnerable populations, such as people with intellectual disability and dementia, represent a high-risk group across multiple dimensions, including a higher prevalence of pre-existing conditions, lower health maintenance, and a propensity for rapid community spread. This led to an urgent need for reliable in-house rapid testing to be performed prior to hospital admission. In the present study, we describe a pooling procedure in which oropharyngeal…
Differential expression of two glucocorticoid receptors in seabass (teleost fish) head kidney after exogeneous cortisol inoculation
2009
Stressful conditions include a prompt release of corticosteroid hormones which can mediate gene expression through glucocorticoid receptors (GR). Since two seabass (Dicentrarchus labrax) GRs have been cloned and sequenced from peritoneal cavity cells (DlGR1) and liver (DlGR2), a comparative amino acid sequence analysis that included Haplochromis burtoni HbGRs, was carried out and homologies disclosed. The DlGR1 and DlGR2 deduced aminoacid sequences showed 61% identity (I) and 70% similarity (S). Moreover, DlGR2 was similar to HbGR2b (69% I, 73% S), and the DlGR1 to HbGR1 (72% I, 78% S). In addition, we examined the expression of the DlGRs after exogeneous cortisol inoculation into the perit…