964 resultados para MT-PCR
Resumo:
Welsch (Projektbearbeiter): Bekanntgabe der Mandatierung des Tünchermeisters Schmidtill als Vertreter der Erlanger Gewerbetreibenden auf dem 'Deutschen Handwerker- und Gewerbe-Congreß' zu Frankfurt/Main
Resumo:
The present report describes a real-time PCR-based procedure to reliably determine the quantity of Leishmania amastigotes in relation to the amount of host tissue in histological skin sections from canine and equine cases of cutaneous leishmaniasis. The novel diagnostic Leishmania-PCR has a detection limit of <0.02 amastigotes per μg tissue, which corresponds well to the detection limit of immunohistochemistry and is far beyond that of conventional histology. Our results emphasise the importance of PCR to complement routine histology of cutaneous leishmaniasis cases, particularly in laboratories in which no immunohistochemical assay is available.
Resumo:
Serial quantification of BCR-ABL1 mRNA is an important therapeutic indicator in chronic myeloid leukaemia, but there is a substantial variation in results reported by different laboratories. To improve comparability, an internationally accepted plasmid certified reference material (CRM) was developed according to ISO Guide 34:2009. Fragments of BCR-ABL1 (e14a2 mRNA fusion), BCR and GUSB transcripts were amplified and cloned into pUC18 to yield plasmid pIRMM0099. Six different linearised plasmid solutions were produced with the following copy number concentrations, assigned by digital PCR, and expanded uncertainties: 1.08±0.13 × 10(6), 1.08±0.11 × 10(5), 1.03±0.10 × 10(4), 1.02±0.09 × 10(3), 1.04±0.10 × 10(2) and 10.0±1.5 copies/μl. The certification of the material for the number of specific DNA fragments per plasmid, copy number concentration of the plasmid solutions and the assessment of inter-unit heterogeneity and stability were performed according to ISO Guide 35:2006. Two suitability studies performed by 63 BCR-ABL1 testing laboratories demonstrated that this set of 6 plasmid CRMs can help to standardise a number of measured transcripts of e14a2 BCR-ABL1 and three control genes (ABL1, BCR and GUSB). The set of six plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (https://ec.europa.eu/jrc/en/reference-materials/catalogue/; CRM code ERM-AD623a-f).
Resumo:
Background. Large field studies in travelers' diarrhea (TD) in multiple destinations are limited by the need to perform stool cultures on site in a timely manner. A method for the collection, transport and storage of fecal specimens that does not require immediate processing, refrigeration and is stable for months would be advantageous. ^ Objectives. Determine if enteric pathogen bacterial DNA can be identified in cards routinely used for evaluation of fecal occult blood. ^ Methods. U.S. students traveling to Mexico in 2005-07 were followed for occurrence of diarrheal illness. When ill, students provided a stool specimen for culture and occult blood by the standard method. Cards were then stored at room temperature prior to DNA extraction. A multiplex fecal PCR was performed to identify enterotoxigenic Escherichia coli and enteroaggregative E. coli (EAEC) in DNA extracted from stools and occult blood cards. ^ Results. Significantly more EAEC cases were identified by PCR done in DNA extracted from cards (49%) or from frozen feces (40%) than by culture followed by HEp-2 adherence assays (13%). Similarly more ETEC cases were detected in card DNA (38%) than fecal DNA (30%) or culture followed by hybridization (10%). Sensitivity and specificity of the card test was 75% and 62%, respectively, and 50% and 63%, respectively, when compared to EAEC and ETEC culture, respectively, and 53% and 51%, respectively compared to EAEC multiplex fecal PCR and 56% and 70%, respectively, compared to ETEC multiplex fecal PCR. ^ Conclusions. DNA extracted from fecal cards used for detection of occult blood is of use in detecting enteric pathogens. ^
Resumo:
Microsatellite instability (MSI) is a hallmark of the mutator phenotype associated with Hereditary Non-Polyposis Colon Cancer (HNPCC). The MSI-High (MSI-H) HNPCC population has been well characterized, but the microsatellite low and stable (MSI-L/MSS) HNPCC population is much less understood. We hypothesize there are significant levels of MSI in HNPCC DNA classified as MSI-L/MSS, but no single variant allele makes up a sufficient population in the tumor DNA to be detected by standard analysis. Finding variants would suggest there is a mutator phenotype for the MSI-L/MSS HNPCC population that is distinct from the MSI-H HNPCC populations. This study quantified and compared MSI in HNPCC patients previously shown to be MSI-H, MSI-L/MSS and an MSI-H older, sporadic colorectal cancer patient. Small-pool Polymerase Chain Reactions (SP-PCRs) were conducted where the DNAs from each sample and controls are diluted into multiple pools, each containing approximately single genome equivalents. At least 100 alleles/sample were studied at six microsatellite loci. Mutant fragments were identified, quantified, and compared using Poisson statistics. Most of the variants were small deletions or insertions, with more mutants being deletions, as has been previously described in yeast and transgenic mice. SP-PCR, where most of the pools contained only 3 or less fragments, enabled identification of variants too infrequent to be detected by large pool PCR. Mutant fragments in positive control MSI-H tumor samples ranged from 0.26 to 0.68 in at least 4 of the 6 loci tested and were consistent with their MSI-H status. In the so called MSS tumors and constitutive tissues (normal colon tissue, and PBLs) of all the HNPCC patients, low, but significant levels of MSI were seen in at least two of the loci studied. This phenomenon was not seen in the sporadic MSI constitutive tissues nor the normal controls and suggests haploinsufficiency, gain-of-function, or a dominant/negative basis of the instability in HNPCC patients carrying germline mutations for tumor suppressor genes. A different frequency and spectrum of mutant fragments suggests a different genetic basis (other than a major mutation in MLH1 or MSH2) for disease in MSI-L and MSS HNPCC patients. ^
Resumo:
The taxonomy of Antarctic fishes has been predominantly based on morphological characteristics rather than on genetic criteria. A typical example is the Notothenia group, which includes N. coriiceps Richardson, 1844, N. neglecta Nybelin, 1951 and N. rossii Richardson, 1844. The Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) technique was used to determine whether N. coriiceps Richardson, 1844 and N. neglecta Nybelin, 1951 are different or whether they are the same species with morphological, physiological and behavioural variability. N. rossii was used as control. Mitochondrial DNA (mtDNA) was isolated from muscle specimens of N. coriiceps Richardson, 1844, N. neglecta Nybelin, 1951 and N. rossii, which were collected in Admiralty Bay, King George Island. The DNA was used to amplify a fragment (690 base pairs) of the mitochondrial gene coding region of NADH dehydrogenase subunit 2. Further, the amplicon was digested with the following restriction enzymes: DdeI, HindIII and RsaI. The results showed a variation of the digestion pattern of the fragment amplified between N. rossii, and N. coriiceps Richardson, 1844 or N. neglecta Nybelin, 1951. However, no differences were found between N. coriiceps Richardson, 1844 and N. neglecta Nybelin, 1951, on the grounds of the same genetic pattern shown by the two fish.
Resumo:
El problema estudiado son las representaciones de género y su relación con la pobreza que se ponen en juego en la interacción docente-alumno. Utilizamos la estrategia de investigación acción-participativa que no es común en investigación social pero ofrece muchas herramientas a los protagonistas para repensar su práctica. Consistió en un estudio de caso único de una escuela de Virreyes que atiende a niños en condiciones de pobreza. Como instrumentos de recolección utilizamos entrevistas en profundidad con docentes y observaciones de clase. Luego realizamos un taller con docentes buscando problematizar participativamente la práctica explicitando los supuestos sobre los que se trabaja. Concluimos que existen ciertas transformaciones en las ideas tradicionales de género, pero aún no han permeado la práctica docente. Encontramos en las aulas estudiadas, y en el discurso de los docentes, ideas estereotipadas sobre varones y mujeres que se ponen en acto en la interacción con sus alumnos. Los docentes, en general, ignoran el carácter simbólico, cultural y arbitrario de sus ideas sobre género. La investigación participativa permitió poner en juego componentes teóricos y de acción para cuestionar estas ideas, llevando a cabo un proceso de reflexión y transformación conjunta de la práctica de los docentes protagonistas del estudio
Resumo:
El problema estudiado son las representaciones de género y su relación con la pobreza que se ponen en juego en la interacción docente-alumno. Utilizamos la estrategia de investigación acción-participativa que no es común en investigación social pero ofrece muchas herramientas a los protagonistas para repensar su práctica. Consistió en un estudio de caso único de una escuela de Virreyes que atiende a niños en condiciones de pobreza. Como instrumentos de recolección utilizamos entrevistas en profundidad con docentes y observaciones de clase. Luego realizamos un taller con docentes buscando problematizar participativamente la práctica explicitando los supuestos sobre los que se trabaja. Concluimos que existen ciertas transformaciones en las ideas tradicionales de género, pero aún no han permeado la práctica docente. Encontramos en las aulas estudiadas, y en el discurso de los docentes, ideas estereotipadas sobre varones y mujeres que se ponen en acto en la interacción con sus alumnos. Los docentes, en general, ignoran el carácter simbólico, cultural y arbitrario de sus ideas sobre género. La investigación participativa permitió poner en juego componentes teóricos y de acción para cuestionar estas ideas, llevando a cabo un proceso de reflexión y transformación conjunta de la práctica de los docentes protagonistas del estudio
Resumo:
In February of 1983 a new terrestrial photogrammetric survey of Lewis Glacier (0° 9' S) has been made, from which the present topographic map has been produced in a scale of 1:5000. Simultaneously a survey of 1963 was evaluated giving a basis for computations of area and volume changes over the 20 year period: Lewis Glacier has lost 22 % of its area and 50 % of its volume. Based on maps and field observations of moraines 10 different stages were identified. Changes of area and volume can be determined for the periods after 1890, two older, undated stages are presumed to be of Little Ice Age-origin. Moderate losses from 1890 to 1920 were followed by strong, uninterrupted retreat up to present. In this respect Lewis Glacier behaves as all other equatorial glaciers that were closer examined. Compared to alpine glaciers the development was similar up to 1950. In the following years, however, the glaciers of the Alps gained mass and advanced while Lewis Glacier experienced its strongest losses from 1974 to 1983.
Resumo:
A high-resolution, 8000 year-long ice core record from the Mt. Logan summit plateau (5300 m asl) reveals the initiation of trans-Pacific lead (Pb) pollution by ca. 1730, and a >10-fold increase in Pb concentration (1981-1998 mean = 68.9 ng/l) above natural background (5.6 ng/l) attributed to rising anthropogenic Pb emissions from Asia. The largest rise in North Pacific Pb pollution from 1970-1998 (end of record) is contemporaneous with a decrease in Eurasian and North American Pb pollution as documented in ice core records from Greenland, Devon Island, and the European Alps. The distinct Pb pollution history in the North Pacific is interpreted to result from the later industrialization and less stringent abatement measures in Asia compared to North America and Eurasia. The Mt. Logan record shows evidence for both a rising Pb emissions signal from Asia and a trans-Pacific transport efficiency signal related to the strength of the Aleutian Low.