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Sample records for vph mediante pcr

  1. Detección de VPH en boca y cérvix de pacientes con diagnóstico citológico sugestivo de infección genital HPV detection in the mouth and cervix of patients with histological diagnosis suggestive of genital infection

    OpenAIRE

    Z. De Guglielmo; M. Ávila; D. Veitía; A. Fernandes; C. Venegas; M. Correnti de Plata

    2012-01-01

    En este trabajo se evaluó la presencia de VPH en la cavidad bucal (mediante oroscopia y citología oral exfoliativa) y su relación con la infección genital en mujeres con diagnóstico citológico sugestivo de infección por VPH. La muestra consistió en 60 pacientes a quienes se les realizó oroscopia, citología y determinación viral en boca y cérvix por PCR, utilizando los iniciadores genéricos MY09/MY11 y MPCR. Se detectó ADN de VPH en las mucosas oral y genital en 48,33 % y 73,3% de las paciente...

  2. Genotipificación de aislamientos de Bartonella bacilliformis por amplificación de elementos repetitivos mediante el uso de REP-PCR y ERIC-PCR

    Directory of Open Access Journals (Sweden)

    Carlos Padilla R

    2003-07-01

    Full Text Available Objetivos: Genotipificar los aislamientos de Bartonella bacilliformis a través de la amplificación de elementos repetitivos mediante el uso de ERIC-PCR y REP-PCR, y determinar si existe variabilidad genética entre aislamientos de varias zonas endémicas. Materiales y Métodos: Se evaluaron mediante el uso del ERIC-PCR y REP-PCR 17 aislamientos de B. bacilliformis de Lima, Cusco y Ancash. Los aislamientos fueron realizados durante los años 1998 y 1999. Para el análisis de los patrones de bandas se usó el software GelCompar 4,0. Resultados: Fueron identificados en los 17 aislamientos 10 genotipos. Los genotipos D, E y H fueron detectados en Cusco; mientras que los genotipos B, C, G, J e I en Lima; y el genotipo F en Ancash. Conclusiones: Nuestros resultados sugieren que REP-PCR y ERIC-PCR son métodos útiles para genotipificar aislamientos de B. bacilliformis. La variabilidad genética debe ser tomada en cuenta en estudios epidemiológicos y clínicos de Bartonelosis; así como el desarrollo de nuevas técnicas diagnósticas y de vacunas.

  3. Curved VPH gratings for novel spectrographs

    Science.gov (United States)

    Clemens, J. Christopher; O'Donoghue, Darragh; Dunlap, Bart H.

    2014-07-01

    The introduction of volume phase holographic (VPH) gratings into astronomy over a decade ago opened new possibilities for instrument designers. In this paper we describe an extension of VPH grating technology that will have applications in astronomy and beyond: curved VPH gratings. These devices can disperse light while simultaneously correcting aberrations. We have designed and manufactured two different kinds of convex VPH grating prototypes for use in off-axis reflecting spectrographs. One type functions in transmission and the other in reflection, enabling Offnerstyle spectrographs with the high-efficiency and low-cost advantages of VPH gratings. We will discuss the design process and the tools required for modelling these gratings along with the recording layout and process steps required to fabricate them. We will present performance data for the first convex VPH grating produced for an astronomical spectrograph.

  4. Las 47 preguntas sobre el virus del papiloma humano, VPH

    Directory of Open Access Journals (Sweden)

    Elena de la Fuente Díez

    2008-09-01

    Full Text Available La prevención del cáncer de cuello uterino ha evolucionado rápidamente a consecuencia de la identificación de su causa, el Virus del Papiloma Humano (VPH. La asociación VPH/Cáncer de Cuello Uterino, es aplicable tanto al carcinoma escamocelular como al adenocarcinoma. Dos de los tipos oncogénicos, los VPHs 16 y 18, son responsables del 70% de los casos de cáncer de cuello uterino a nivel mundial. El VPH es muy prevalente entre personas sexualmente activas, y se puede identificar fácilmente mediante el uso de tecnologías capaces de detectar ADN y otros biomarcadores del VPH. El ADN y los biomarcadores se pueden detectar en todas las fases de la historia natural de la infección y del proceso neoplásico. Dicha trazabilidad unida a la validación clínica de las tecnologías de identificación, han permitido mejorar los protocolos de cribado del cáncer cervical. La proteína principal del virión, ya expresada, produce inmunógenos capaces de inducir fuertes respuestas inmunitarias cuando se administran vía intramuscular, y ésta es la base de la Vacuna frente a VPH.The UCC (Uterine Cervical Cancer prevention has suffered a great evolution because of the knowing of its cause, wich is the HPV (Human Papillomavirus. The association HPV / UCC is available also to Scamocellular Carcinome to Adenocarcinome. Mundially, the oncogenic types, 16 and 18, are responssible of 70% of cases of UCC. HPV is very prevalent in sexually active people, and it’s easy identify by using DNA amplyfiyng technologies and other biomarkers. These ones can be detected in hole virus natural life, including the neoplasic process. The tazability and the clinic validation of this technologies, have improved to get a better screening of the UCC. The main HPV protein, is able to produce a strong inmunitary response, when it’s given by intramuscular punctury; and this is the explanation of the HPC vaccine.

  5. Towards distributed multiscale computing for the VPH

    NARCIS (Netherlands)

    Hoekstra, A.G.; Coveney, P.

    2010-01-01

    Multiscale modeling is fundamental to the Virtual Physiological Human (VPH) initiative. Most detailed three-dimensional multiscale models lead to prohibitive computational demands. As a possible solution we present MAPPER, a computational science infrastructure for Distributed Multiscale Computing

  6. Vacuna contra el VPH (HPV Vaccine)

    Centers for Disease Control (CDC) Podcasts

    2013-07-25

    Desde el 2006, hay una vacuna que protege contra los tipos de VPH que causan cáncer con mayor frecuencia. Este podcast habla sobre la importancia de que los padres hablen con los proveedores de atención médica de sus hijos sobre ponerles la vacuna contra el VPH.  Created: 7/25/2013 by MMWR.   Date Released: 11/4/2013.

  7. Detección rápida de resistencia a drogas en Mycobacterium tuberculosis mediante PCR-SSCP y PCR- Heteroduplex

    Directory of Open Access Journals (Sweden)

    Róger Calderón E

    2003-04-01

    Full Text Available Objetivo: Detectar tempranamente la susceptibilidad a las drogas antituberculosas rifampicina e isoniacida mediante PCR y electroforesis conformacional. Materiales y métodos: Se implementaron dos ensayos de amplificación de los genes rpoB y katG y mediante Heteroduplex y SSCP se determinó la susceptibilidad antituberculosa de 31 muestras clínicas procedentes de pacientes con diagnóstico de tuberculosis pulmonar baciloscopía positiva. La caracterización fenotípica de la susceptibilidad, se realizó empleando el método de las proporciones. Resultados: Los ensayos de PCR detectaron hasta 2,5 pg de ADN genómico de M. tuberculosis; no amplificando ADN de otras micobacterias y bacterias comunes de la flora bucal. Se encontró una concordancia general entre la detección molecular y convencional de la susceptibilidad a rifampicina e isoniacida de 96,7% y 83,9% (p<0,05, respectivamente. Sin embargo, sólo en pacientes con antecedente de tratamiento se presentó una concordancia del 100% y 90,9% (p<0,05 para rifampicina e isoniacida, respectivamente. Además, este sistema de detección de resistencia puede emitir resultados 48 horas después de la recepción de la muestra clínica. Conclusiones: Estos sistemas se presentan como una excelente alternativa para la identificación temprana de pacientes infectados con bacilos de M. tuberculosis drogoresistentes. Potencialmente, se podrán dirigir óptimos y oportunos esquemas terapéuticos que contribuirán con el control y prevención de la transmisión de cepas multidrogo-resistentes que afectan en gran medida a la salud pública de nuestro país.

  8. Detección del virus de la leucosis bovina en ganado criollo colombiano mediante PCR-anidado

    Directory of Open Access Journals (Sweden)

    Darwin Yovanny Hernández-Herrera

    2011-12-01

    Full Text Available Se evaluó la presencia del virus de la leucosis bovina (VLB en 360 muestras de ADN de ocho razas bovinas criollas: Blanco Orejinegro (BON, Casanareño (CAS, Costeño con Cuernos (CCC, Chino Santandereano (ChS, Caqueteño (CQT, Hartón del Valle (HV, Romosinuano (RS y San Martinero (SM, dos Razas Sintéticas Colombianas: Lucerna (LUC y Velásquez (VEL y dos razas foráneas: Brahmán (B y Holstein (H. Para la detección del pro-virus se amplificó una región del gen env viral, mediante PCR anidada. La presencia del VLB fue mayor en la raza HV seguido por ChS (83.3% y 60% respectivamente, VEL y LUC tuvieron el mismo porcentaje (50%, en CAS, CCC y CQT la presencia del virus fue de 26.7%, 23.3% y 16.7% respectivamente; no se encontró el virus en BON, SM y RS. En las razas foráneas la presencia fue de 83.3% para H y 6.7% para B. Se encontró dependencia altamente significativa entre la presencia del VLB y la raza, el sexo y región de origen de la muestra. El promedio de presencia en las razas criollas fue menor que en las foráneas, menor en los machos que en las hembras y en la región norte que en el suroccidente y el centro del país.

  9. Evaluación de las técnicas de detección del VPH en los programas de cribado para cáncer de cuello uterino Assessment of hpv detection assays for use in cervical cancer screening programs

    Directory of Open Access Journals (Sweden)

    M Paz Cañadas

    2006-10-01

    Full Text Available OBJETIVO: La identificación de la infección por tipos de alto riesgo del virus del papiloma humano (VPH es una herramienta útil para el cribado de cáncer del cuello uterino. Las distintas técnicas aplicadas para su detección deben contrastarse y validarse para su empleo en la tamización poblacional. MATERIAL Y MÉTODOS: Se evalúan tres técnicas para la detección del VPH en 166 muestras cervicales procedentes de mujeres atendidas en una clínica de dermatología en Oviedo (España: a PCR-EIA mediante consensos MY09/MY011; b PCR con line blot hybridization (PCR-LBH con consensos PGMY; y c hybrid capture 2. RESULTADOS: El ADN-VPH se reconoció en 29.5%, 25.3% y 24.7%, de acuerdo con el ensayo. La concordancia global entre PCR-EIA, PCR-LBH y HC2 fue de 73.5% con los valores de kappa superiores a 0.56 entre los ensayos (pOBJECTIVE: Detection of high-risk human papillomavirus types (HPV infection is an important tool in the screening of cervical cancer and triage of cytological abnormalities. The different techniques for detection of this cancer need to be contrasted and validated for use in population screening. MATERIAL AND METHODS: Cervical cell samples were collected from 166 women attending a dermatology clinic in Oviedo (Spain. We evaluated the performance of three different assays for VPH detection. The methods utilized were 1 In-house PCR-EIA using L1 consensus primers MY09/MY11, 2 A PCR-reverse line blot hybridization (PCR-LBH that uses L1 consensus PGMY primers. 3 Hybrid Capture 2. All assays were performed blinded. The kappa statistic was used to test for global agreement between assay pairs. RESULTS: HPV DNA was detected in 24,7%, 25,3% and 29,5% of the women, respective to the assay. The overall agreement between the in-house PCR, PCR-LBH and HC2 was (73.5% with all kappa values between assay pairs exceeding 0.56 (p<0.001. CONCLUSION: The three HPV assays were equally accurate in estimating high-risk HPV prevalence and HPV

  10. Data Management Services for VPH Applications

    CERN Multimedia

    CERN. Geneva

    2014-01-01

    The VPH-Share project [1] develops a cloud platform for the Virtual Physiological Human (VPH) research community. One of the key challenges is to share and access large datasets used by medical applications to transform them into meaningful diagnostic information. VHP researchers need advanced storage capabilities to enable collaboration without introducing additional complexity to the way data are accessed and shared. In the VPH-Share cloud platform [2], the data storage federation [3] is achieved by an aggregation of data resources in a client-centric manner and exposing it via a standardized protocol that can be also mounted and presented as a local storage so a kind of a file system abstraction is provided. There is a common management layer that uses loosely coupled and independent storage resources and with such a layer a variety of storage resources such as simple file servers, storage clouds and data grid may be aggregated exposing all available storage. As a result, distributed applications have ...

  11. Metrology measurements for large-aperture VPH gratings

    Science.gov (United States)

    Zheng, Jessica R.; Gers, Luke; Heijmans, Jeroen

    2013-09-01

    The High Efficiency and Resolution Multi Element Spectrograph (HERMES) for the Australian Astronomical Observatory (AAO) uses four large aperture, high angle of incidence volume phase holographic gratings (VPHG) for high resolution `Galactic archaeology' spectroscopy. The large clear aperture, the high diffraction efficiency, the line frequency homogeneity, and mosaic alignment made manufacturing and testing challenging. We developed new metrology systems at the AAO to verify the performance of these VPH gratings. The measured diffraction efficiencies and line frequency of the VPH gratings received so far meet the vendor's provided data. The wavefront quality for the Blue VPH grating is good but the Green and Red VPH gratings need to be post polishing.

  12. PCR

    African Journals Online (AJOL)

    Elham

    2013-07-03

    Jul 3, 2013 ... was constructed with competitive strategy by PCR-cloning technique and the limitation range was determined. The PCR products of MTB and IAC were 245 and 660 bp, respectively on .... products' differentiation was easy.

  13. Las 47 preguntas sobre el virus del papiloma humano, VPH

    OpenAIRE

    Elena de la Fuente Díez; Luz María Mira Ferrer

    2008-01-01

    La prevención del cáncer de cuello uterino ha evolucionado rápidamente a consecuencia de la identificación de su causa, el Virus del Papiloma Humano (VPH). La asociación VPH/Cáncer de Cuello Uterino, es aplicable tanto al carcinoma escamocelular como al adenocarcinoma. Dos de los tipos oncogénicos, los VPHs 16 y 18, son responsables del 70% de los casos de cáncer de cuello uterino a nivel mundial. El VPH es muy prevalente entre personas sexualmente activas, y se puede identificar fácilmente m...

  14. CARACTERIZACIÓN MOLECULAR MEDIANTE rep-PCR DE AISLADOS NATIVOS DE Bacillus thuringiensis, OBTENIDOS DE MUESTRAS DE SUELO

    Directory of Open Access Journals (Sweden)

    Fabián Galvis

    2014-01-01

    Full Text Available Bacillus thuringiensis es una bacteria Gram-positiva formadora de esporas, que produ - ce cristales parasporales de naturaleza proteica, tóxicos contra diferentes órdenes de insectos y biodegradables e inocuos para otras especies. Esta investigación empleó el modelo experimen - tal, que mediante técnicas de observación permi - tió, la identificación microbiológica y bioquímica de B. thuringiensis a partir de muestras de suelo de los municipios de Cúcuta, El Zulia, Los Patios, San Cayetano y Villa del Rosario, Norte de Santander, Colombia, y su posterior caracteri - zación con los marcadores moleculares Bc-Rep y MB1. Se identificaron microbiológica y bioquí - micamente 10 aislados como B. thuringiensis ; los resultados del análisis filogenético mostraron diferencias significativas en los agrupamientos obtenidos con los marcadores Bc-Rep y MB1. Con Bc-Rep se registró un índice de similaridad bajo (18%, mientras que con el marcador MB1 se obtuvo un índice mayor de similitud, 58%. En este trabajo se evidenció una gran variabilidad genética entre los aislados, que mostraron a los marcadores Bc-Rep y MB1 como altamente efectivos para diferenciar cepas estrechamente relacionadas, convirtiéndose en una herramienta genética de gran valor para estudios de identifi-cación y diversidad en B. thuringiensis.

  15. Tipificación capsular mediante PCR de aislamientos de Haemophilus influenzae no tipificables por aglutinación PCR-based capsular typing of Haemophilus influenzae isolates non-typeable by agglutination

    Directory of Open Access Journals (Sweden)

    G. Weltman

    2005-12-01

    Full Text Available Haemophilus influenzae es reconocido como un agente patógeno responsable de infecciones localizadas y sistémicas. Se han descrito 6 tipos de polisacáridos capsulares antigénicamente distintos (a, b, c, d, e, y f que se pueden identificar por aglutinación en lámina con antisueros específicos. También existen cepas no capsuladas (NC fenotípicamente no tipificables (NT. La introducción de la vacuna conjugada produjo una marcada disminución de las enfermedades invasivas causadas por H. influenzae tipo b. En este contexto, la tipificación capsular mediante PCR es el método más apropiado para distinguir las cepas no capsuladas de las mutantes b deficientes en cápsula (b- y detectar la presencia de cepas pertenecientes a otros serotipos que no puedan ser tipificables por aglutinación. Se determinó el genotipo capsular a 38 aislamientos de Haemophilus influenzae no tipificables por aglutinación, derivados al servicio de Bacteriología Clínica del INEI-ANLIS "Dr. Carlos G. Malbrán" en el período 2002-2004. El 78,9% de los aislamientos provenían de hemocultivos y la mayor parte de ellos estaban asociados a foco respiratorio. El 100% de los aislamientos fueron identificados como H. influenzae no capsulados mediante la técnica de PCR.Haemophilus influenzae is recognized as a pathogenic agent responsible of localized and systemic infections. Six antigenically different capsular polysaccharide types have been described (a, b, c, d, e, and f which can be identified by slide agglutination with specific antisera. Besides there are non capsulated strains that cannot be typed by slide agglutination. The introduction of the conjugated vaccine produced an important reduction of invasive diseases caused by H. influenzae type b. Capsular typing by PCR is the most appropriated method for distinguishing non capsulated strains from capsule deficient type b mutants (b- and for detecting strains of other serotypes that cannot be detected by slide

  16. Incidencia, duración y determinantes de la infección por virus de papiloma humano (VPH en una cohorte de mujeres colombianas

    Directory of Open Access Journals (Sweden)

    Mónica Molano

    2004-03-01

    Full Text Available

    Ciertos tipos de VPH son los agentes etiológicos de cáncer cervical, lesiones tempranas y ulceras genitales (1-4. Las infecciones por VPH tienen alta prevalencia y se transmiten por contacto sexual (5-6. Los datos publicados sobre incidencia son en mujeres jóvenes pero son escasos los datos en mujeres de mediana edad (7-10. Se sugiere que los VPH de alto riesgo tienen mayor duración que los de bajo riesgo (7-11.

    Para examinar estos aspectos se presentan los datos sobre una cohorte de 2,200 mujeres seguidas desde Noviembre de 1993 a julio del 2001 en Bogotá, sexualmente activas, entre 13-85 años de edad. Se realizó detección y tipificación del VPH mediante PCR estándar GP5+/GP6+ e inmunoensayo enzimático (EIA. Para el cálculo de la incidencia se usaron métodos de persona-tiempo, asumiendo que la infección se presentó en la mitad de periodo entre la última visita negativa a VPH y la siguiente positiva.

    Para este análisis se tomaron 1,610 mujeres después de excluir las infecciones prevalentes. La duración media de seguimiento por paciente fue de 4,1 años (Rango intercuartil RIQ 3.2-5.0, la mediana de seguimiento fue de 7 meses (RIQ 6.0-12.0, más del 65% de las mujeres tenían 4 visitas y la edad media fue de 32,3 años (RIQ 26,5-39,2. La incidencia para infección por VPH fue de 6,2 por 100 personas-año, siendo la incidencia mayor para los tipos de alto riesgo que para los de bajo riesgo.

    Los tipos de mayor incidencia fueron los VPH 16, 58, 31 y 18. La curva de incidencia por edad mostró una forma bimodal, con un pico de incidencia de 15-19 años, que decrece hasta alrededor de los 50 años, cuando presenta un pequeño incremento

  17. Toward a VPH/Physiome ToolKit.

    Science.gov (United States)

    Garny, Alan; Cooper, Jonathan; Hunter, Peter J

    2010-01-01

    The Physiome Project was officially launched in 1997 and has since brought together teams from around the world to work on the development of a computational framework for the modeling of the human body. At the European level, this effort is focused around patient-specific solutions and is known as the Virtual Physiological Human (VPH) Initiative.Such modeling is both multiscale (in space and time) and multiphysics. This, therefore, requires careful interaction and collaboration between the teams involved in the VPH/Physiome effort, if we are to produce computer models that are not only quantitative, but also integrative and predictive.In that context, several technologies and solutions are already available, developed both by groups involved in the VPH/Physiome effort, and by others. They address areas such as data handling/fusion, markup languages, model repositories, ontologies, tools (for simulation, imaging, data fitting, etc.), as well as grid, middleware, and workflow.Here, we provide an overview of resources that should be considered for inclusion in the VPH/Physiome ToolKit (i.e., the set of tools that addresses the needs and requirements of the Physiome Project and VPH Initiative) and discuss some of the challenges that we are still facing.

  18. Detección y cuantificación del Potato mop-top virus (PMTV en Colombia mediante qRT-PCR

    Directory of Open Access Journals (Sweden)

    Nevar García Bastidas

    2013-04-01

    Full Text Available El Potato mop-top virus (PMTV es uno de los virus re-emergentes en cultivos de papa en Colombia. Es transmitido por Spongospora subterranea, el agente causal de la sarna polvosa. La detección del PMTV presenta dificultades debido a su distribución irregular en las plantas, bajo título y movimiento sistémico como ARN desnudo. Con el fin de ampliar el rango de herramientas disponibles para detectar el PMTV en los programas de certificación de tubérculo-semilla, en este estudio se evaluó la prueba de RT-PCR en tiempo real (qRT-PCR en dos pasos: con los cebadores PMTV-1948F/PMTV-2017R y la sonda Taqman® PMTV-1970, dirigidos al gen CP-RT del ARN2 viral. Se construyó una curva estándar a partir de la transcripción in vitro de un fragmento de 1513 pb de este gen. Posteriormente, se evaluó la utilidad de la técnica a partir de tres tipos de muestras: plantas señuelo de Nicotiana benthamiana y Solanum phureja inoculadas con quistosoros de Sss, raíces de papa con síntomas de sarna polvosa del municipio de La Unión (Antioquia y tubérculos-semilla. Mediante qRT-PCR fue posible detectar el virus en 11 de las 20 muestras de raíz de plantas señuelo, mientras que 14 de las 15 muestras de raíces de papa resultaron positivas, estimándose una concentración entre 4.72 x 10(11 y 7.60 x 10(13 partículas virales/µl. Adicionalmente, en el ensayo de tubérculo-semilla se determinó la presencia del PMTV en una de las 16 muestras. Estos resultados indican la viabilidad de utilizar rutinariamente la técnica de qRT-PCR para la detección de PMTV en Colombia.

  19. Support for Taverna workflows in the VPH-Share cloud platform.

    Science.gov (United States)

    Kasztelnik, Marek; Coto, Ernesto; Bubak, Marian; Malawski, Maciej; Nowakowski, Piotr; Arenas, Juan; Saglimbeni, Alfredo; Testi, Debora; Frangi, Alejandro F

    2017-07-01

    To address the increasing need for collaborative endeavours within the Virtual Physiological Human (VPH) community, the VPH-Share collaborative cloud platform allows researchers to expose and share sequences of complex biomedical processing tasks in the form of computational workflows. The Taverna Workflow System is a very popular tool for orchestrating complex biomedical & bioinformatics processing tasks in the VPH community. This paper describes the VPH-Share components that support the building and execution of Taverna workflows, and explains how they interact with other VPH-Share components to improve the capabilities of the VPH-Share platform. Taverna workflow support is delivered by the Atmosphere cloud management platform and the VPH-Share Taverna plugin. These components are explained in detail, along with the two main procedures that were developed to enable this seamless integration: workflow composition and execution. 1) Seamless integration of VPH-Share with other components and systems. 2) Extended range of different tools for workflows. 3) Successful integration of scientific workflows from other VPH projects. 4) Execution speed improvement for medical applications. The presented workflow integration provides VPH-Share users with a wide range of different possibilities to compose and execute workflows, such as desktop or online composition, online batch execution, multithreading, remote execution, etc. The specific advantages of each supported tool are presented, as are the roles of Atmosphere and the VPH-Share plugin within the VPH-Share project. The combination of the VPH-Share plugin and Atmosphere engenders the VPH-Share infrastructure with far more flexible, powerful and usable capabilities for the VPH-Share community. As both components can continue to evolve and improve independently, we acknowledge that further improvements are still to be developed and will be described. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Frecuencia de las mutaciones más comunes del gen CFTR en pacientes peruanos con fibrosis quística mediante la técnica ARMS-PCR

    OpenAIRE

    Aquino, Ruth; Protzel, Ana; Rivera, Juan; Abarca, Hugo; Dueñas, Milagros; Nestarez, Cecilia; Purizaga, Nestor; Diringer, Benoit

    2017-01-01

    Objetivos. Determinar la frecuencia de las diez mutaciones más comúnmente reportadas en América Latina del gen CFTR mediante Sistema de Mutación Refractario a la amplificación por PCR (ARMS-PCR) en los pacientes con fibrosis quística (FQ) de dos instituciones hospitalarias de referencia en el Perú durante el año 2014. Materiales y métodos. Se evaluó la frecuencia de las diez comúnmente reportadas más comúnmente reportadas del gen CFTR en los pacientes del Hospital Nacional Edgardo Rebagliati ...

  1. Frecuencia de microsporidiosis intestinal en pacientes positivos para VIH mediante las técnicas de Gram cromotropo rápido y PCR.

    Directory of Open Access Journals (Sweden)

    Jorge H. Botero

    2004-12-01

    Full Text Available Los microsporidios son protozoos intracelulares obligados, implicados en procesos de diarrea persistente en pacientes con sida, aunque no son exclusivos de este grupo de pacientes. La prevalencia de microsporidios en diferentes países varía entre 8% y 52%. En nuestro medio no se conoce su frecuencia, por lo que este trabajo se propuso determinar la frecuencia de microsporidiosis intestinal en pacientes positivos para VIH, mediante la prueba del Gram cromotropo rápido (quick hot Gram y la PCR; para esto se realizó un estudio prospectivo, descriptivo, con una población intencional de todos los pacientes positivos para VIH remitidos al Laboratorio del Grupo Interdisciplinario para el Estudio de las Parasitosis Intestinales por las diferentes instituciones de atención de pacientes positivos para VIH de Medellín en el periodo comprendido entre agosto de 2001 y septiembre de 2002. Se hizo una encuesta clínico-epidemiológica y se practicaron análisis coprológicos seriados que incluían examen directo, por concentración y tinciones especiales para coccidias y microsporidios intestinales; además, se solicitó recuento de linfocitos TCD4+ y carga viral. Se estudiaron 103 pacientes en edades comprendidas entre 2 y 74 años; el 70% (72/103 presentaba diarrea al ingreso al estudio; la mayoría (83,5% fueron hombres. La frecuencia global de microsporidiosis intestinal fue de 3,9% (4/103; se encontraron tres pacientes positivos para Enterocytozoon bieneusi y uno con Encephalitozoon intestinalis; otras parasitosis intestinales representaron el 39,8%. La frecuencia de microsporidiosis en este estudio fue relativamente baja; además, como era de esperarse, la mayoría de los casos de microsporidios estuvieron asociados con diarrea prolongada y recuentos de LTCD4+ menores de 100 cél/?l y cargas virales superiores a 100.000 copias (3/4.

  2. Polimorfismo genético de beta-lactoglobulina y alphalactoalbúmina en el ganado criollo colombiano, mediante PCR-SSCP

    Directory of Open Access Journals (Sweden)

    Jaime A Rosero-Alpala

    2011-12-01

    Full Text Available La población de ganado criollo colombiano ha venido presentando una inquietante disminución al pasar de 23.415 ejemplares en 1999 a 20.102 en 2003. A pesar de los esfuerzos por recuperar las razas criollas el panorama para su conservación es incierto, por tanto la búsqueda de caracteres deseables puede contribuir a su valoración y conservación. Los genes relacionados con el mejoramiento de la calidad de la leche producida por estas razas se consideran de gran importancia en la industria láctea, por tal razón y con el objetivo de caracterizar los genes beta-lactoglobulina y alpha-lactoalbúmina se analizaron 30 muestras de sangre de cada una de las razas criollas (Blanco Orejinegro, Caqueteño, Casanareño, Costeño con cuernos, Chino Santandereano, Hartón del Valle, Romosinuano y Sanmartinero, dos razas sintéticas colombianas (Lucerna y Velásquez y dos razas foráneas (Holstein y Brahman. Se amplificaron fragmentos de 262pb para beta-lactoglobulina (b-LG y de 166 pb para alpha-lactoalbúmina (a-LA que se genotipificaron mediante PCR-SSCP. El promedio de la frecuencia para b-LG A y b-LG B fue de 0.46 ± 0.020 y de 0.53 ± 0.020, respectivamente, y de 0.35 ± 0.019 para a-LA A y 0.64 ± 0.019 para a-LA B. El promedio de diversidad genética (He para b-LG fue 0.498 y de 0.455 para a-LA. Los ganados criollos representan una base genética valiosa, como alternativa para mejorar genéticamente los hatos destinados a la producción de leche con mejores características en calidad para la industria láctea.

  3. Polimorfismo genético de beta-lactoglobulina y alphalactoalbúmina en el ganado criollo colombiano, mediante PCR-SSCP

    Directory of Open Access Journals (Sweden)

    Muñoz Florez Jaime Eduardo

    2011-12-01

    Full Text Available La población de ganado criollo colombiano ha venido presentando una inquietante disminución al pasar de 23.415 ejemplares en 1999 a 20.102 en 2003. A pesar de los esfuerzos por recuperar las razas criollas el panorama para su conservación es incierto, por tanto la búsqueda de caracteres deseables puede contribuir a su valoración y conservación. Los genes relacionados con el mejoramiento de la calidad de la leche producida por estas razas se consideran de gran importancia en la industria láctea, por tal razón y con el objetivo de caracterizar los genes beta-lactoglobulina y alpha-lactoalbúmina se analizaron 30 muestras de sangre de cada una de las razas criollas (Blanco Orejinegro, Caqueteño, Casanareño, Costeño con cuernos, Chino Santandereano, Hartón del Valle, Romosinuano y Sanmartinero, dos razas sintéticas colombianas (Lucerna y Velásquez y dos razas foráneas (Holstein y Brahman. Se amplificaron fragmentos de 262pb para beta-lactoglobulina (b-LG y de 166 pb para alpha-lactoalbúmina (a-LA que se genotipificaron mediante PCR-SSCP. El promedio de la frecuencia para b-LG A y b-LG B fue de 0.46 ± 0.020 y de 0.53 ± 0.020, respectivamente, y de 0.35 ± 0.019 para a-LA A y 0.64 ± 0.019 para a-LA B. El promedio de diversidad genética (He para b-LG fue 0.498 y de 0.455 para a-LA. Los ganados criollos representan una base genética valiosa, como alternativa para mejorar genéticamente los hatos destinados a la producción de leche con mejores características en calidad para la industria láctea.

  4. Detección del virus de la leucosis bovina en ganado criollo colombiano mediante PCR-anidado Bovine leukemia virus detection in Creole Colombian breeds using nested-PCR

    Directory of Open Access Journals (Sweden)

    Darwin Yovanny Hernández-Herrera

    2011-12-01

    Full Text Available Se evaluó la presencia del virus de la leucosis bovina (VLB en 360 muestras de ADN de ocho razas bovinas criollas: Blanco Orejinegro (BON, Casanareño (CAS, Costeño con Cuernos (CCC, Chino Santandereano (ChS, Caqueteño (CQT, Hartón del Valle (HV, Romosinuano (RS y San Martinero (SM, dos Razas Sintéticas Colombianas: Lucerna (LUC y Velásquez (VEL y dos razas foráneas: Brahmán (B y Holstein (H. Para la detección del pro-virus se amplificó una región del gen env viral, mediante PCR anidada. La presencia del VLB fue mayor en la raza HV seguido por ChS (83.3% y 60% respectivamente, VEL y LUC tuvieron el mismo porcentaje (50%, en CAS, CCC y CQT la presencia del virus fue de 26.7%, 23.3% y 16.7% respectivamente; no se encontró el virus en BON, SM y RS. En las razas foráneas la presencia fue de 83.3% para H y 6.7% para B. Se encontró dependencia altamente significativa entre la presencia del VLB y la raza, el sexo y región de origen de la muestra. El promedio de presencia en las razas criollas fue menor que en las foráneas, menor en los machos que en las hembras y en la región norte que en el suroccidente y el centro del país.Using 360 DNA samples from eight Creole bovine breeds Blanco Orejinegro (BON, Casanareño (CAS, Costeño con Cuernos (CCC, Chino Santandereano (ChS, Caqueteño (CQT, Hartón del Valle (HV, Romosinuano (RS and San Martinero (SM, two synthetic Colombian breeds: Lucerna (LUC and Velásquez (VEL and two introduced breeds Brahmán (B and Holstein (H; the presence of Bovine Leukemia Virus (BLV was evaluated through the amplification of a viral gene region env (provirus detection - nested-PCR. The percentage of presence and independence test were calculated (X². Presence of BLV was higher in HV breed, followed by ChS (83.3% and 60% respectively; VEL and LUC breeds showed the same percentage (50%. In CAS, CCC and CQT the presence of virus was 26.7%, 23.3% y 16.7% respectively. On the other hand, no virus presence was

  5. Polimorfismo genético de beta-lactoglobulina y alphalactoalbúmina en el ganado criollo colombiano, mediante PCR-SSCP Genetic polymorphism of beta-lactoglobulin and alpha-lactoalbumin in Colombian Creole cattle by PCR-SSCP

    Directory of Open Access Journals (Sweden)

    Jaime A Rosero-Alpala

    2011-12-01

    Full Text Available La población de ganado criollo colombiano ha venido presentando una inquietante disminución al pasar de 23.415 ejemplares en 1999 a 20.102 en 2003. A pesar de los esfuerzos por recuperar las razas criollas el panorama para su conservación es incierto, por tanto la búsqueda de caracteres deseables puede contribuir a su valoración y conservación. Los genes relacionados con el mejoramiento de la calidad de la leche producida por estas razas se consideran de gran importancia en la industria láctea, por tal razón y con el objetivo de caracterizar los genes beta-lactoglobulina y alpha-lactoalbúmina se analizaron 30 muestras de sangre de cada una de las razas criollas (Blanco Orejinegro, Caqueteño, Casanareño, Costeño con cuernos, Chino Santandereano, Hartón del Valle, Romosinuano y Sanmartinero, dos razas sintéticas colombianas (Lucerna y Velásquez y dos razas foráneas (Holstein y Brahman. Se amplificaron fragmentos de 262pb para beta-lactoglobulina (b-LG y de 166 pb para alpha-lactoalbúmina (a-LA que se genotipificaron mediante PCR-SSCP. El promedio de la frecuencia para b-LG A y b-LG B fue de 0.46 ± 0.020 y de 0.53 ± 0.020, respectivamente, y de 0.35 ± 0.019 para a-LA A y 0.64 ± 0.019 para a-LA B. El promedio de diversidad genética (He para b-LG fue 0.498 y de 0.455 para a-LA. Los ganados criollos representan una base genética valiosa, como alternativa para mejorar genéticamente los hatos destinados a la producción de leche con mejores características en calidad para la industria láctea.The Colombian Creole Cattle has showed a preoccupant population decreasing, from 23,415 individuals in 1999 to 20,102 in 2003. Despite that many efforts to recover the creole breeds have been done, its future conservation is unclear. Searching for economic desirable genes may contribute to its preservation and utilization as a genetic resource. Genes related with the improvement of milk proteins are considered as an economic important

  6. Translation of vph mRNA in Streptomyces lividans and Escherichia coli after removal of the 5' untranslated leader.

    Science.gov (United States)

    Wu, C J; Janssen, G R

    1996-10-01

    The Streptomyces vinaceus viomycin phosphotransferase (vph) mRNA contains an untranslated leader with a conventional Shine-Dalgarno homology. The vph leader was removed by ligation of the vph coding sequence to the transcriptional start site of a Streptomyces or an Escherichia coli promoter, such that transcription would initiate at the first position of the vph start codon. Analysis of mRNA demonstrated that transcription initiated primarily at the A of the vph AUG translational start codon in both Streptomyces lividans and E. coli; cells expressing the unleadered vph mRNA were resistant to viomycin indicating that the Shine-Dalgarno sequence, or other features contained within the leader, was not necessary for vph translation. Addition of four nucleotides (5'-AUGC-3') onto the 5' end of the unleadered vph mRNA resulted in translation initiation from the vph start codon and the AUG triplet contained within the added sequence. Translational fusions of vph sequence to a Tn5 neo reporter gene indicated that the first 16 codons of vph coding sequence were sufficient to specify the translational start site and reading frame for expression of neomycin resistance in both E. coli and S. lividans.

  7. DETECCIÓN Y CUANTIFICACIÓN DE Spongospora subterranea f. sp. subterranea EN PLANTAS SEÑUELO Y CULTIVOS DE PAPA EN COLOMBIA MEDIANTE qPCR

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    Nevar Alirio García Bastidas

    2013-01-01

    Full Text Available La sarna polvosa de la papa (Solanum tuberosum , S. phureja causada por Spongospora subterranea f. sp. subterranea (Sss, es una de las enfermedades más limitantes de este cultivo. En Colombia, se han empleado diferentes métodos de detección asintomática de Sss, incluyendo bioensayos con plantas señuelo, PCR de ITS y pruebas de ELISA. Sin embargo, sus niveles de sensibilidad son bajos o requieren tiempos extensos. Una alternativa para complementar dichas herramientas es la PCR cuantitativa en tiempo real (qPCR. En este trabajo se evaluó dicha técnica utilizando los juegos de cebadores SsTQF1-SsTQR1; Spon421F-Spon494R y SscolF-SscolR (diseñados en este estudio, bajo la metodología de SYBR Green®; mientras que con Taqman® se evaluaron los cebadores SponF-SponR y la sonda SponP. Una vez determinada la funcionalidad de los cebadores, se descartó por inespecificidad, el par Spon421F-Spon494R; para los restantes se realizaron curvas estándar basadas en diluciones seriadas de quistosoros. Las pruebas de qPCR detectaron a Sss en las 20 muestras evaluadas de plantas señuelo de Nicotiana benthamiana y papa, utilizando los cebadores SsTQF1-SsTQR1 (Ct: 10,57- 29,34 y SscolF-SscolR (Ct: 14,39-34,08; mientras que 19 de las muestras fueron positivas con SponF-SponR-SponP (Ct: 15,63-38,93. A partir de 20 muestras de raíces de papa de cultivos de La Unión (Antioquia, Colombia, fue posible detectar el patógeno en 17 de ellas con SscolF-SscolR, estimándose una concentración de 6470 a 1,39 x 1010 quistorosos/mL. Estos resultados indican la ocurrencia de altos niveles de inóculo de Sss en esta región y enfatizan en la necesidad de fortalecer los programas de certificación de tubérculo-semilla en Colombia.

  8. Detección de Listeria spp. y Listeria monocytogenes en muestras de leche cruda y quesos artesanales respectivamente, mediante PCR en Tiempo Real

    Directory of Open Access Journals (Sweden)

    Viviana Pamela Chiluisa-Utreras

    2017-07-01

    Full Text Available Background. In Ecuador, studies about bacteria genre Listeria in artisanal cheeses are scarce, and in raw milk, practically nonexistent. Milk production is one of the main livestock activities in the province of Pichincha and it is essential to study these products. Since all the cantons that make up Pichincha are milk producers, three of them, Cayambe, Quito and Pedro Moncayo were randomly sampled. Objective. To determine Listeria spp. And Listeria monocytogenes in samples of raw milk and artisanal cheeses, respectively, using Real Time PCR. Methods. The application of the qPCR technique in the detection of microorganisms and especially of bacteria in food, is based on four fundamental aspects: its sensitivity, specificity, speed and processing capacity of large sample flow. It is possible to detect small amounts of pathogenic microorganisms, such as Listeria spp in raw milk, after extraction and quantification of total DNA. Results. In this study in raw milk, one positive was determined from a total of 60 samples, representing 1.6% of Listeria spp. and 16 positive samples of 45, representing 35.6% of Listeria monocytogenes in artisanal cheeses from three farms in the province of Pichincha. Conclusions. The results, according to the statistical analyzes carried out with the Kruskal - Wallis test, show that in Pichincha the bacterium is present in raw milk, but in non - representative quantities, whereas for Listeria monocytogenes there is statistical significance in the cheeses samples

  9. Características sociodemográficas de mujeres peruanas con virus papiloma humano detectado por PCR-RFLP

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    Yasser Sullcahuaman-Allende

    Full Text Available Con el objetivo de determinar las características sociodemográficas del virus de pacientes con papiloma humano (VPH referidas al Instituto Nacional de Enfermedades Neoplásicas (INEN durante los años 2012-2014, se realizó la detección del VPH en células cervicales por reacción en cadena de la polimerasa (PCR. En 465 muestras cervicales se detectaron 151 (32,5% casos de VPH positivas. Los genotipos más frecuentes fueron VPH-16 (23,8% y VPH-6 (11,9%. La presencia de VPH fue mayor en mujeres de 17 a 29 años (OR 2,64, IC 95%:1,14-6,13 y solteras (OR 2,31, IC 95%: 1,37-3,91, la presencia de genotipos de VPH de alto riesgo fue mayor en solteras (OR 2,19, IC 95%: 1,04-4,62. En conclusión, mujeres jóvenes y solteras presentaron mayor frecuencia de casos VPH-positivos a quienes se debe enfatizar la participación en programas de tamizaje con métodos moleculares y citológicos combinados, a fin de detectar oportunamente el riesgo de desarrollar cáncer de cuello uterino

  10. Quantificazione mediante PCR dell’EBV-DNA da biopsie cutanee di pazienti con linfomi cutanei primitivi (micosi fungoide e sindrome di Sèzary

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    Chiara Merlino

    2007-06-01

    Full Text Available Mycosis fungoides (MF, the most indolent form of CTCL, originates from a clonal expansion of epidermotropic helper/memory T cells. Sezary syndrome (SS is a rare primay epidermotropic cutaneous T-cell lymphoma in leukemic. The aetiopathogenesis of MF and SS remains obscure despite several investigations. Infectious, environmental and genetic factors have been implicated as potential aetiological agents. The studies investigating the role of EBV in CTCL present conflicting results. The different sensitivities of the technical methods used in the evaluation of the presence of viral DNA or virus-related antigens make comparison of the results difficult. The aim of this study was to retrospectively evaluate the EBV-DNA load in skin biopsies from MF and SS patients by a highly sensitive (1-10 EBV-DNA copies/reaction quantitative-competitive PCR (QC-PCR developed in our lab to better asses the relationship between EBV and CTCL. Skin biopsies were obtained from 21 MF and 10 SS patients; skin biopsies from a 8 patients with inflammatory skin disease were used as controls. EBV-DNA was detected in 70% of biopsies from SS patients vs. 0% of MF patients. No control patients resulted EBV-DNA positive, as expected. In addition, in SS patients, the survival from diagnosis is lesser in EBV-positive patients vs.EBV-negative patients even if not statistically significant.We are going to investigate the presence of EBV-DNA in peripheral blood of a larger number of patients and to evaluate the pattern of viral genes expression, to better assess the aetiopathogenetical role of EB virus in this kind of neoplasies.

  11. Authentication of beef, carabeef, chevon, mutton and pork by a PCR-RFLP assay of mitochondrial cytb gene

    OpenAIRE

    Kumar, Deepak; Singh, S. P.; Karabasanavar, Nagappa S.; Singh, Rashmi; Umapathi, V.

    2012-01-01

    Authentication of meat assumes significance in view of religious, quality assurance, food safety, public health, conservation and legal concerns. Here, we describe a PCR-RFLP (Polymerase Chain Reaction- Restriction Fragment Length Polymorphism) assay targeting mitochondrial cytochrome-b gene for the identification of meats of five most common food animals namely cattle, buffalo, goat, sheep and pig. A pair of forward and reverse primers (VPH-F & VPH-R) amplifying a conserved region (168–776 b...

  12. Conocimientos sobre el virus del papiloma humano (VPH) y su vacuna dentro de la comunidad universitaria

    OpenAIRE

    San Martín González, Paula

    2016-01-01

    Objetivo: Evaluar el nivel de conocimientos sobre el VPH en una muestra de universitarios. Se realizó un estudio observacional descriptivo y analítico de corte transversal con el objetivo principal de evaluar el nivel de conocimientos y actitudes sobre VPH en una muestra de universitarios de la Universidad Pontificia de Salamanca. La muestra cuenta con 194 sujetos, 103 estudiantes del grado de enfermería y 91 estudiantes del grado de psicología que cumplían los criterios de inclusión y ...

  13. Papiloma bucal producido por VPH y su relación con carcinoma

    OpenAIRE

    Martínez Martínez, Adel; Baldiris Ávila, Rosa; Díaz Caballero, Antonio

    2012-01-01

    El virus del papiloma humano (VPH) es el responsable de múltiples manifestaciones en boca, las cuales generalmente se caracterizan por lesiones vegetantes, verrugosidades o lesiones papulares. La incidencia de la enfermedad en boca ha aumentado ya que el contagio por vía sexual es una de las principales vías de contagio, lo que también permite que cada vez exista un mayor número de diagnósticos de VPH oncogénico en cavidad bucal. The human papiloma virus (HPV) is responsible for multiple m...

  14. Presence of a Phytoplasma Associated with Witches’-Broom Disease in Ugni molinae Turcz. and Gaultheria phillyreifolia (Pers. Sleumer Determined by DAPI, PCR, and DNA Sequencing Presencia de un Fitoplasma Asociado a la Enfermedad de "Escoba de Bruja" en Ugni molinae Turcz. y Gaultheria phillyreifolia (Pers. Sleumer Determinado Mediante DAPI, PCR y Secuenciación de ADN

    Directory of Open Access Journals (Sweden)

    Nolberto Arismendi S

    2010-03-01

    Full Text Available Murta (Ugni molinae Turcz. and common chaura (Gaultheria phillyreifolia (Pers. Sleumer are native species of Chile. Plants of both species have shown over-branching like witches' broom. The causal agents of these symptoms in many plants are phytoplasma. To verify the presence of these microorganisms, DAPI (4',6-diamidino-2-phenylindole staining analysis and polymerase chain reaction (PCR were performed in symptomatic and asymptomatic plants. Positive PCR samples were sequenced to identify the pathogens involved. In individuals of both species with witches’ broom symptoms, DAPI staining showed fluorescent bodies in the phloem tissues, but not in asymptomatic plants. Verification by nested-PCR, phytoplasmatic DNA was amplified from diseased murta and chaura, but not in apparently healthy plants. Sequencing of amplified products allowed locating phytoplasma within the ash yellows group (16SrVII and related to Candidatus phytoplasma fraxini. This is the first report of phytoplasma in Chilean native species. Considering the diversity of plant species infected by the ash yellows group suggests that G. phillyreifolia and U. molinae could be a phytoplasma reservoir for other economically important agricultural crops.La murta (Ugni molinae Turcz. y la chaura común (Gaultheria phillyreifolia (Pers. Sleumer son especies nativas de Chile. En plantas de ambas especies se ha observado una sobre-ramificación de tipo "escoba de bruja". En muchas plantas los agentes causales de esta sintomatología son fitoplasmas. Para verificar la presencia de estos microorganismos se analizaron plantas con y sin síntomas mediante tinciones DAPI (4’,6-diamidino-2-fenilindol y reacción en cadena de la polimerasa (PCR. Muestras positivas en la PCR fueron secuenciadas para identificar al fitopatógeno implicado. En individuos de ambas especies con síntomas de escoba de bruja, la tinción DAPI permitió observar cuerpos fluorescentes en los tejidos del floema, situaci

  15. Comparing modelling techniques when designing VPH gratings for BigBOSS

    Science.gov (United States)

    Poppett, Claire; Edelstein, Jerry; Lampton, Michael; Jelinsky, Patrick; Arns, James

    2012-09-01

    BigBOSS is a Stage IV Dark Energy instrument based on the Baryon Acoustic Oscillations (BAO) and Red Shift Distortions (RSD) techniques using spectroscopic data of 20 million ELG and LRG galaxies at 0.5VPH) gratings have been identified as a key technology which will enable the efficiency requirement to be met, however it is important to be able to accurately predict their performance. In this paper we quantitatively compare different modelling techniques in order to assess the parameter space over which they are more capable of accurately predicting measured performance. Finally we present baseline parameters for grating designs that are most suitable for the BigBOSS instrument.

  16. Detección de toxoplasmosis congénita en líquido amniótico humano mediante la técnica de nested-pcr

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    A. Hortúa

    2000-07-01

    Full Text Available La toxoplasmosis es provocada por el parasite intracelular obligado Toxoplasma gondii,de la familia Toxoplasmidae (Flores, 1991. Este parasite puede ser asintomático en adultos con un sistema inmune normal, pero puede ser de gran trascendencia en el feto en gestación y en pacientes con SIDA o deficiencia en el sistema inmune (Montoya, 1996. La presencia de anticuerpos antitoxoplasma indica únicamente que la persona se infecto con el microorganismo en un momento dado y no que haya oeste desarrollando la toxoplasmosis necesariamente, pero un resultado positivo indica que el individuo está en riesgo de desarrollar la enfermedad (Perea, 1983. \\ Si la infección se produce durante el embarazo, existe la posibilidad que la toxoplasmosis sea transmitida al feto ocasionando aborto espontaneo, prematuridad o enfermedades severas en el feto, tales como: hidrocefalia y calcificaciones inn-ace- i rebrales (Picazo, 1994. En la mayoría de los casos el diagnóstico biológico de la toxoplasmosis congénita se basa en métodos serológicos indirectos; sin embargo, en los últimos años los diversos estudios realizados en Biología Molecular permitieron utilizar la Técnica de Reacción en Cadena de la Polimerasa (PCR para el diagnóstico de la enfermedad (Hohlfeld, 1994. Los primeros estudios en PCR fueron dirigidos a la amplificación de la secuencia repetitiva del gen B1 de Toxoplasma gondii en líquido amniótico de mujeres infectadas (Grover, 1990. La prueba de PCR en liquido amniótico es definitivamente mas sensible que otras técnicas convencionales usadas, ya que estas presentan dificultad en establecer un diagnóstico segura y oportuno, por esto se ha implementando la técnica de PCR en la detección de la toxoplasmosis, aportando un progreso indiscutible en aquellos casos donde los exámenes clínicos y serológicos presentan limitaciones. También disminuye el tiempo de análisis de las muestras arrojando resultados en un período máximo de 24

  17. ESTUDIO DE LA DIVERSIDAD GENÉTICA DE 20 ACCESIONES DE CACAO (Theobroma cacao L. MEDIANTE AP-PCR DE LA COLECCIÓN DEL CENTRO DEL CACAO DE AROMA TENGUEL EN LA FINCA EXPERIMENTAL LA BUSETA

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    Mercedes Susana Carranza Patiño

    2008-06-01

    total (HT obtenida para las 20 accesiones de T. cacao mediante RAPD fue de 0.636.

  18. Detección mediante RT-PCR en tiempo real del virus vacunal en cerdos inmunizados con la vacuna cubana contra la peste porcina clásica

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    Tania Campos-Cuello

    2016-08-01

    Full Text Available La peste porcina clásica (PPC es una enfermedad viral infectocontagiosa, producida por un virus ARN del género Pestivirus, familia Flaviviridae. En la actualidad es una de las causas de pérdidas económicas en la industria porcina a nivel mundial. En su prevención se han utilizado vacunas vivas atenuadas, empleando la cepa China lapinizada. La Reacción en Cadena de la Polimerasa Reverso Transcriptasa (RT-PCR ha sido uno de los métodos más sensibles aplicado en Medicina Veterinaria para la detección de virus ARN. En el caso del virus de la PPC es muy útil porque el ácido nucleico se puede detectar desde muy temprano en la infección y en periodos más largos en aquellos animales que se recuperan. El objetivo de este estudio fue aplicar la técnica de RT-PCR en tiempo real para la detección de la cepa China lapinizada de la vacuna cubana contra la PPC. Las tonsilas de los cerdos vacunados fueron el órgano más positivo en la detección del ARN del virus vacunal. Los resultados obtenidos evidenciaron una interferencia del virus vacunal en el diagnóstico, siendo el día 12 posvacunación en el que se obtiene una emisión umbral de fluorescencia más bajo.

  19. DETECCIÓN Y DIFERENCIACIÓN DE Mycoplasma gallisepticum Y Mycoplasma synoviae MEDIANTE LA TÉCNICA DE PCR A PARTIR DE HISOPOS TRAQUEALES DE AVES CON SÍNTOMAS RESPIRATORIOS

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    CESAR VENTURA

    2012-09-01

    Full Text Available Los micoplasmas son importantes patógenos en las aves por ser responsables de cuadros respiratorios que ocasionan grandes pérdidas económicas a la industria avícola a nivel mundial. Existen principalmente dos especies de micoplasmas como causantes de enfermedad en aves comerciales, el Mycoplasma gallisepticum (MG y el Mycoplasma synoviae (MS. Teniendo en cuenta su importancia y la necesidad de conocer y diferenciar la presencia de las diferentes especies de micoplasmas presentes en las explotaciones avícolas, se tomaron 91 muestras de hisopos traqueales de aves con síntomas respiratorios, provenientes de igual número de granjas de pollo de engorde, ponedoras comerciales y reproductoras pesadas ubicadas en los departamentos de Cundinamarca y Boyacá y se determinó la presencia de MG y MS por la técnica de PCR. La prevalencia determinada fue de 39,6% para MG y 47,3% para MS, encontrándose diferencias estadísticamente significativas cuando se comparó la positividad a MG y MS y el tipo de explotación (p

  20. vph6 mutants of Saccharomyces cerevisiae require calcineurin for growth and are defective in vacuolar H(+)-ATPase assembly.

    Science.gov (United States)

    Hemenway, C S; Dolinski, K; Cardenas, M E; Hiller, M A; Jones, E W; Heitman, J

    1995-11-01

    We have characterized a Saccharomyces cerevisiae mutant strain that is hypersensitive to cyclosporin A (CsA) and FK506, immunosuppressants that inhibit calcineurin, a serine-threonine-specific phosphatase (PP2B). A single nuclear mutation, designated cev1 for calcineurin essential for viability, is responsible for the CsA-FK506-sensitive phenotype. The peptidyl-prolyl cis-trans isomerases cyclophilin A and FKBP12, respectively, mediate CsA and FK506 toxicity in the cev1 mutant strain. We demonstrate that cev1 is an allele of the VPH6 gene and that vph6 mutant strains fail to assemble the vacuolar H(+)-ATPase (V-ATPase). The VPH6 gene was mapped on chromosome VIII and is predicted to encode a 181-amino acid (21 kD) protein with no identity to other known proteins. We find that calcineurin is essential for viability in many mutant strains with defects in V-ATPase function or vacuolar acidification. In addition, we find that calcineurin modulates extracellular acidification in response to glucose, which we propose occurs via calcineurin regulation of the plasma membrane H(+)-ATPase PMA1. Taken together, our findings suggest calcineurin plays a general role in the regulation of cation transport and homeostasis.

  1. Detection of the mosquitocidal toxin genes encoding Cry11 proteins from Bacillus thuringiensis using a novel PCR-RFLP method Detección de genes que codifican proteínas mosquitocidas Cry11 de Bacillus thuringiensis mediante un método de PCR-RFLP novedoso

    Directory of Open Access Journals (Sweden)

    D. H. Sauka

    2010-02-01

    Full Text Available A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP method for detection of cry11 genes from Bacillus thuringiensis was established. Based on the analysis of conserved regions of the cry11 genes, 2 oligonucleotide primers were designed to amplify a 1459-bp fragment of the cry11Aa gene, and a 1471-bp of the cry11Ba and cry11Bb genes. The amplification products were digested with restriction endonuclease HinfI. Exotic B. thuringiensis strains and native isolates collected from soils, leaves and stored product dust of Argentina were analyzed to study the distribution of cry11 genes. The PCR-RFLP patterns revealed the detection of cry11 genes in 3 of 64 exotic strains and in 10 of 107 native B. thuringiensis isolates tested. Just the cry11Aa gene subclass was detected among these bacteria. Since the methodology was also developed to detect cry11Ba and cry11Bb genes, an experimental future confirmation will be required. Based on the results obtained, the PCR-RFLP method presented may be a valuable tool for specific detection of the mosquitocidal toxin genes encoding Cry11 proteins from B. thuringiensis.En el presente estudio se estableció una estrategia basada en la amplificación génica (PCR y el posterior análisis de restricción (RFLP para detectar todos los genes cry11 de Bacillus thuringiensis informados hasta ahora. De acuerdo con el análisis de las regiones conservadas en los genes cry11, se diseñaron dos cebadores para amplificar un fragmento de 1459 pb de los genes cry11Aa y un fragmento de 1471 pb de los genes cry11Ba y cry11Bb. Los productos de la amplificación fueron digeridos con la enzima de restricción HinfI. Se analizaron cepas exóticas de B. thuringiensis y aislamientos nativos de Argentina obtenidos a partir de muestras de suelos, hojas y polvillo de silos, para estudiar la distribución de los genes cry11. Los patrones de PCR-RFLP revelaron la presencia de genes cry11 en 3 de las 64 cepas ex

  2. Infección por VPH en mujeres del municipio de Pasto (Colombia) con resultados de citología normal

    OpenAIRE

    Sánchez-Ortega, Claudia; Suárez, Narváez Karen; Yépez-Chamorro, Maria; Guerrero-Flórez, Milena

    2013-01-01

    Introducción. El cáncer de cuello uterino (CCU) asociado a la infección por VPH se considera un importante problema de salud pública a nivel mundial. En Colombia, es la segunda causa de muerte por cáncer en mujeres. En el municipio de Pasto (N) ubicado al suroccidente de Colombia, la tasa de incidencia es de 27,3/100.000 habitantes. Profundizar en la historia natural de la infección por VPH permitiría mejorar las estrategias de control y diagnóstico temprano para evitar la progresión de este ...

  3. Aceptabilidad y conocimientos sobre la vacunación contra el virus del papiloma humano (VPH) en médicos ginecólogos de la Argentina

    OpenAIRE

    Alejandro Mazzadi; Melisa Paolino; Silvina Arrossi

    2012-01-01

    OBJETIVO: Evaluar entre los ginecólogos argentinos la aceptabilidad y prescripción de la vacuna contra el virus del papiloma humano (VPH), los conocimientos sobre sus características y uso, y las nociones médico-biológicas sobre infección por VPH y cáncer cervicouterino. MATERIAL Y MÉTODOS: Entre noviembre de 2009 y marzo de 2010 se encuestaron a 686 ginecólogos vía internet. RESULTADOS: Más de 80% de los encuestados prescribe la vacuna, conoce sus características y administración, y consider...

  4. OpenCMISS: a multi-physics & multi-scale computational infrastructure for the VPH/Physiome project.

    Science.gov (United States)

    Bradley, Chris; Bowery, Andy; Britten, Randall; Budelmann, Vincent; Camara, Oscar; Christie, Richard; Cookson, Andrew; Frangi, Alejandro F; Gamage, Thiranja Babarenda; Heidlauf, Thomas; Krittian, Sebastian; Ladd, David; Little, Caton; Mithraratne, Kumar; Nash, Martyn; Nickerson, David; Nielsen, Poul; Nordbø, Oyvind; Omholt, Stig; Pashaei, Ali; Paterson, David; Rajagopal, Vijayaraghavan; Reeve, Adam; Röhrle, Oliver; Safaei, Soroush; Sebastián, Rafael; Steghöfer, Martin; Wu, Tim; Yu, Ting; Zhang, Heye; Hunter, Peter

    2011-10-01

    The VPH/Physiome Project is developing the model encoding standards CellML (cellml.org) and FieldML (fieldml.org) as well as web-accessible model repositories based on these standards (models.physiome.org). Freely available open source computational modelling software is also being developed to solve the partial differential equations described by the models and to visualise results. The OpenCMISS code (opencmiss.org), described here, has been developed by the authors over the last six years to replace the CMISS code that has supported a number of organ system Physiome projects. OpenCMISS is designed to encompass multiple sets of physical equations and to link subcellular and tissue-level biophysical processes into organ-level processes. In the Heart Physiome project, for example, the large deformation mechanics of the myocardial wall need to be coupled to both ventricular flow and embedded coronary flow, and the reaction-diffusion equations that govern the propagation of electrical waves through myocardial tissue need to be coupled with equations that describe the ion channel currents that flow through the cardiac cell membranes. In this paper we discuss the design principles and distributed memory architecture behind the OpenCMISS code. We also discuss the design of the interfaces that link the sets of physical equations across common boundaries (such as fluid-structure coupling), or between spatial fields over the same domain (such as coupled electromechanics), and the concepts behind CellML and FieldML that are embodied in the OpenCMISS data structures. We show how all of these provide a flexible infrastructure for combining models developed across the VPH/Physiome community. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Valor del ADN-VPH en el cribado de la población oportunista en el departamento 6 de Valencia

    OpenAIRE

    Mora Moya, María

    2012-01-01

    La infecció pel Virus del Papil.loma Humà (VPH) és la causa principal de gairebé tots els casos de càncer cervical. En països on s'apliquen de manera programada tècniques de detecció, hi ha una disminució de la incidència i la mortalitat del càncer cervical. La detecció del VPH és un avenç important per a la prevenció del càncer en permetre un diagnòstic precoç de lesions cancerígenes. És per això que, es va decidir estudiar durant un any a la població de pacients que acudien a les consultes ...

  6. Prevalencia de infección por virus de papiloma humano (VPH de alto riesgo y factores asociados en embarazadas derechohabientes del IMSS en el estado de Morelos The prevalence of high-risk HPV infection in pregnant women from Morelos, México

    Directory of Open Access Journals (Sweden)

    Carlos Hernández-Girón

    2005-12-01

    Full Text Available OBJETIVO: Algunos estudios sugieren la posibilidad de que el proceso fisiológico del embarazo modifique algunas características del hospedero, lo que incrementa el riesgo de infección por VPH; sin embargo, esta asociación no está bien establecida. Pocos estudios se han realizado para determinar la prevalencia de infección por VPH de alto riesgo en mujeres embarazadas, y sus factores relacionados. El presente estudio busca determinar la prevalencia de infección por VPH de alto riesgo, en una muestra de mujeres embarazadas mexicanas, y sus posibles factores de riesgo. MATERIAL Y MÉTODOS: Se realizó un estudio epidemiológico de tipo transversal en una muestra de 274 mujeres embarazadas que acudieron a los servicios de primer nivel de atención del Hospital General del Instituto Mexicano del Seguro Social (IMSS en Cuernavaca, Morelos, durante el año 2000. Se obtuvieron muestras de exudado vaginal mediante autotoma, y se aplicó un cuestionario estructurado sobre características sociodemográficas, ginecoobstétricas y de comportamiento sexual. La detección de infección por VPH de alto riesgo, se realizó empleando un método de captura de híbridos (Hybrid Capture II, HCII, Digene Corp.. RESULTADOS: La prevalencia de infección por VPH fue de 37.2% (102/274. En promedio acudieron a su primera cita prenatal al sexto mes de embarazo; la media de edad fue 25.7 años. Los principales factores de riesgo asociados a infección por VPH fueron: edad, entre 20 y 29 años (RM = 2.82; IC95% 1.02-7.76, 30 o más años (RM ajustada = 6.85; IC95% 1.22-38.2; compañeros sexuales con otras parejas (RM= 2.05; IC95% 1.2-3.7. Mostraron asociación positiva, aunque marginalmente significativas: escolaridad menor de 6 años (RM = 1.67; IC95% 0.67-4.3; más de dos parejas sexuales en su vida (RM = 1.54; IC95% 0.7-3.4; y tabaquismo actual (RM= 1.6; IC95% 0.6-5.0. CONCLUSIONES: Los hallazgos indican una mayor prevalencia de infección por VPH de alto riesgo

  7. Aceptabilidad y conocimientos sobre la vacunación contra el virus del papiloma humano (VPH en médicos ginecólogos de la Argentina

    Directory of Open Access Journals (Sweden)

    Alejandro Mazzadi

    2012-10-01

    Full Text Available OBJETIVO: Evaluar entre los ginecólogos argentinos la aceptabilidad y prescripción de la vacuna contra el virus del papiloma humano (VPH, los conocimientos sobre sus características y uso, y las nociones médico-biológicas sobre infección por VPH y cáncer cervicouterino. MATERIAL Y MÉTODOS: Entre noviembre de 2009 y marzo de 2010 se encuestaron a 686 ginecólogos vía internet. RESULTADOS: Más de 80% de los encuestados prescribe la vacuna, conoce sus características y administración, y considera la necesidad de continuar con el tamizaje cervical en mujeres vacunadas. El 37% posee un conocimiento global de la relación entre vacuna y detección/tratamiento de la patología cervical. De los encuestados, 25% subestima la magnitud de la infección, ≈30% no reconoce el rol etiológico del VPH en la enfermedad, y ≈40% posee un conocimiento global del manejo de la infección. CONCLUSIONES: La aceptabilidad de la vacuna contra el VPH es alta. Debe reforzarse la capacitación de los profesionales sobre vacunación y patología cervical, así como las nociones médico-biológicas sobre infección por VPH y cáncer cervicouterino.OBJECTIVE: To evaluate HPV vaccine acceptability and prescription; knowledge about HPV vaccine; and knowledge about HPV infection and cervical cancer among Argentinean gynecologists. MATERIALS AND METHODS: Between November 2009 and March 2010 we carried out an internet survey of 686 gynecologists. RESULTS: More than 80% of gynecologists prescribed HPV vaccine, knew characteristics of HPV vaccines, and knew that women will still need regular cervical cancer screening after HPV vaccination; 37% had global knowledge about relationship between vaccine, detection and treatment of cervical cancer; 25% underestimated the epidemiological extent of HPV infections, ≈30% was not aware of the causative relationship between HPV infection and cervical cancer and ≈40% had global knowledge about management of HPV infection

  8. Redes sociales de apoyo y género: vivencia de mujeres con VPH, displasias y cáncer cervicouterino

    OpenAIRE

    Castro Vásquez, María del Carmen; Arellano Gálvez, María del Carmen

    2014-01-01

    El objetivo del artículo es analizar el funcionamiento y el tipo de redes sociales de apoyo de mujeres diagnosticadas con infección por el virus del papiloma humano (VPH), displasias y cáncer cervical in situ (Cacu), diferenciando la vivencia por tipo de diagnóstico. Se trata de un estudio cualitativo: se realizaron 34 entrevistas semiestructuradas a mujeres en dos clínicas de displasias en Hermosillo, Sonora. Encontramos que cada diagnóstico tiene connotaciones distintas en la percepción ind...

  9. Risk factors for cervical cancer among HPV positive women in Mexico Factores de riesgo de cáncer cervical en mujeres VPH positivas en México

    Directory of Open Access Journals (Sweden)

    Yvonne N Flores

    2008-02-01

    Full Text Available OBJECTIVE: To identify factors that are associated with an increased risk of developing high-grade cervical intraepithelial neoplasia (CIN or cancer among human papillomavirus (HPV-positive women in Mexico. MATERIAL AND METHODS: A case-control study design was used. A total of 94 cases and 501 controls who met the study inclusion criteria were selected from the 7 732 women who participated in the Morelos HPV Study from May 1999 to June 2000. Risk factor information was obtained from interviews and from HPV viral load results. Odds ratios and 95 percent confidence intervals were estimated using unconditional multivariate regression. RESULTS: Increasing age, high viral load, a young age at first sexual intercourse, and a low socio-economic status are associated with an increased risk of disease among HPV-positive women. CONCLUSIONS: These results could have important implications for future screening activities in Mexico and other low resource countries.OBJETIVO: Identificar factores asociados con un mayor riesgo de desarrollar neoplasia intraepitelial cervical (NIC de alto grado o cáncer en mujeres con virus de papiloma humano (VPH, en México. MATERIAL Y MÉTODOS: Se utilizó un diseño de casos y controles. Un total de 94 casos y 501 controles fueron seleccionados de las 7 732 mujeres que participaron en el Estudio de VPH en Morelos, de mayo de 1999 a junio de 2000. La información sobre factores de riesgo se obtuvo de entrevistas y de los resultados de carga virales de VPH. Se estimaron razones de momios e intervalos de confianza de 95% con modelos multivariados de regresión no condicionada. RESULTADOS: El incremento de edad, la carga viral elevada, la edad temprana al inicio de la vida sexual y el nivel socioeconómico bajo se asocian con un mayor riesgo de enfermedad en mujeres VPH positivas. CONCLUSIONES: Estos resultados podrían tener implicaciones importantes a futuro para las actividades de tamizaje en México y en otros países de

  10. Authentication of beef, carabeef, chevon, mutton and pork by a PCR-RFLP assay of mitochondrial cytb gene.

    Science.gov (United States)

    Kumar, Deepak; Singh, S P; Karabasanavar, Nagappa S; Singh, Rashmi; Umapathi, V

    2014-11-01

    Authentication of meat assumes significance in view of religious, quality assurance, food safety, public health, conservation and legal concerns. Here, we describe a PCR-RFLP (Polymerase Chain Reaction- Restriction Fragment Length Polymorphism) assay targeting mitochondrial cytochrome-b gene for the identification of meats of five most common food animals namely cattle, buffalo, goat, sheep and pig. A pair of forward and reverse primers (VPH-F & VPH-R) amplifying a conserved region (168-776 bp) of mitochondrial cytochrome-b (cytb) gene for targeted species was designed which yielded a 609 bp PCR amplicon. Further, restriction enzyme digestion of the amplicons with Alu1 and Taq1 restriction enzymes resulted in a distinctive digestion pattern that was able to discriminate each species. The repeatability of the PCR-RFLP assay was validated ten times with consistent results observed. The developed assay can be used in routine diagnostic laboratories to differentiate the meats of closely related domestic livestock species namely cattle from buffalo and sheep from goat.

  11. Improving cervical cancer screening in Mexico: results from the Morelos HPV Study Mejorando la detección oportuna del cáncer cervical en México: resultados del Estudio de VPH en Morelos

    Directory of Open Access Journals (Sweden)

    Yvonne Flores

    2003-01-01

    Full Text Available OBJECTIVE: The purpose of this paper is to describe some of the results of the Morelos HPV Study. The main objective of the Morelos HPV Study is to evaluate the use of human papillomavirus (HPV DNA testing, as compared to the Papanicolaou (Pap test, for cervical cancer (CC screening. MATERIAL AND METHODS: The Morelos HPV Study is currently being conducted in Mexico, to examine the possibility of using HPV testing for CC screening. The HPV testing of self-collected vaginal and clinician-collected cervical specimens was evaluated as part of this study. The acceptability of the HPV testing of self-collected specimens was compared to that of the Pap test. A cost-effectiveness analysis (CEA and cost-benefit analysis (CBA was also performed. RESULTS: The Morelos HPV Study results indicate that HPV testing has a greater sensitivity to detect cervical intraepithelial neoplasia (CIN 2/3 and CC than the Pap test. Our results also indicate an over-all lower acceptability of the Pap test as compared to the self-collected procedure. The results of the CEA and CBA indicate that screening women between the ages of 20-80 for CC using some type of HPV testing is always more cost-effective than screening for CC using the Pap test. CONCLUSIONS: Our results suggest that self- and clinician-collected HPV testing could be used in CC prevention programs, as an effective complement or substitute for the Pap test.OBJETIVO: Describir algunos de los resultados del Estudio de VPH en Morelos. El objetivo principal del Estudio de VPH en Morelos es evaluar el uso de la prueba del virus de papiloma humano (VPH, en relación con la prueba de Papanicolaou, para el tamizaje de cáncer cervical. MATERIAL Y MÉTODOS: El Estudio de VPH en Morelos actualmente se está llevando a cabo en México, para examinar la posibilidad de usar la prueba de VPH para la detección de cáncer cervical. Se evaluó el uso de la prueba de VPH en muestras auto-tomadas vaginales y en muestras cervicales

  12. [Combination of etoposide, cisplatin and ifosfamide (VPH) in the salvage chemotherapy of relapsing or refractory aggressive malignant lymphoma. Study of 51 patients].

    Science.gov (United States)

    Eghbali, H; Catry-Thomas, I; Soubeyran, P; Bonnel, C; Hoerni, B

    1994-09-01

    Fifty-one patients with non-Hodgkin's lymphoma refractory or relapsing after CHOP-like regimen, underwent a salvage chemotherapy by VPH: etoposide 100 mg/m2/d, D1 to D3, cisplatin 20 mg/m2/d, D1 to D5, ifosfamide 1 g/m2/d D1 to D5, mesna 1.2 g/m2/d D1 to D5, every 4 weeks. Among 46 evaluable patients for efficacy, 21 (45.6%) achieved complete or partial response according to WHO criteria and 25 (54.3%) failed, while five cases (9.8% of all patients) were not evaluable (two initial complete remission before VPH, two early toxic deaths and one confusional syndrome). Thirty-five patients (68.6%) died of lymphoma, three (5.8%) of acute toxicity and 13 (25.5%) are alive: five in complete remission. The toxicity is mainly myelo-suppression, digestive and renal but could be managed as usually. Although the follow-up is short, this regimen appears effective in these circumstances after CHOP failure but it should be used early, before overt chemoresistance. It does not hinder a bone marrow transplantation programme.

  13. Identificación de Virus Papiloma Humano 16 (vph-16) en carcinoma queratinizante de pulmón IDENTIFICATION OF HUMAN PAPILLOMAVIRUS 16 (HPV 16) IN KERATINIZING LUNG CARCINOMA

    OpenAIRE

    Francisco Aguayo G.; Manuel Meneses M.; Alejandro Corvalán R.; María Luisa Muñoz S.; Chilaya Koriyama; Yoshito Eizuru; Suminori Akiba

    2002-01-01

    El cáncer pulmonar constituye la primera causa de muerte por cáncer en el mundo y la cuarta causa de muerte por cáncer en Chile. El carcinoma escamoso de pulmón representa entre el 35% a 50% de los casos de cáncer pulmonar. Existe fuerte evidencia, aunque aún controversial, respecto de la asociación entre esta forma histológica y la infección por Virus Papiloma Humano (VPH), siendo los genotipos VPH 16 y 18 los que se han asociado a lesiones malignas y premalignas de diversos tejidos epitelia...

  14. HPV vaccine introduction at the local level in a developing country: attitudes and criteria among key actors Introducción de la vacuna contra el VPH en niveles locales de un país en vías de desarrollo: actitudes y criterios de actores clave

    Directory of Open Access Journals (Sweden)

    Marion Piñeros

    2010-05-01

    Full Text Available In most developing countries, HPV vaccines have been licensed but there are no national policy recommendations, nor is it clear how decisions on the introduction of this new vaccine are made. Decentralization processes in many Latin American countries favor decision-making at the local level. Through a qualitative study we explored knowledge regarding the HPV vaccine and the criteria that influence decision-making among local health actors in four regions of Colombia. We conducted a total of 14 in-depths interviews with different actors; for the analysis we performed content analysis. Results indicate that decision-making on the HPV vaccine at the local level has mainly been driven by pressure from local political actors, in a setting where there is low technical knowledge of the vaccine. This increases the risk of initiatives that may foster inequity. Local decisions and initiatives need to be strengthened technically and supported by national-level decisions, guidelines and follow-up.En gran parte de los países en vías de desarrollo, las vacunas contra el VPH tienen aprobación para su comercialización. Sin embargo, no hay recomendaciones y tampoco hay claridad sobre la forma en la que se toman las decisiones para su introducción. La reforma del sistema de salud en muchos países latinoamericanos permite la toma de decisiones en el nivel local. Mediante un estudio cualitativo con actores claves del sector salud en cuatro regiones de Colombia, exploramos el conocimiento sobre la vacuna del VPH y los criterios que influyen en la toma de decisiones. Se realizaron 14 entrevistas en profundidad y análisis de contenido. Los resultados indican que en el nivel local la toma de decisiones sobre la introducción de la vacuna está determinada en gran parte por la presión ejercida por figuras políticas locales. Esto, sumado a un bajo nivel de conocimiento técnico, incrementa la posibilidad de iniciativas con implicaciones éticas considerables

  15. “caracterización de bacterias metalofijadoras de mercurío, a través de la subunidad 16srna, mediante la técnica de pcr-dgge del rio gala (aguas abajo en el recinto San Rafael) en la parroquia tenguel”

    OpenAIRE

    Llivisaca, Susana Alexandra; Burgos, Felix Adolfo; Vargas, Jeffrey David

    2011-01-01

    El proyecto sobre el cual tratamos, se refiere el uso de la técnica molecular PCR-DGGE, para la caracterización de bacterias, y su potencial existencia en las aguas del río Gala de la parroquia Tenguel. La selección del sitio se basó al alto grado de contaminación con metales pesados en la que este se encuentra este rio. La probabilidad sobre la presencia de microorganismos con capacidad metalofijadora, es alta dada sus características biológicas y bioquímicas con las que cuenta esta bacteria...

  16. Terapia anticipada de Aspergilosis Invasora en pacientes oncohematológicos de alto riesgo mediante el uso de PCR para la detección precoz de Aspergillus. Estudio de los polimorfismos genéticos de la vía NFKB y su implicación en las Arpergilosis Invasora

    OpenAIRE

    Villaescusa de la Rosa, María Teresa

    2016-01-01

    [ES] La aspergilosis invasora (AI) es un problema fundamental en el manejo de muchos pacientes hematológicos, por lo que es esencial un diagnóstico precoz que permita instaurar un tratamiento adecuado lo antes posible con el fin de disminuir la morbimortalidad de esta complicación. Nuestra hipótesis es que la asociación del galactomanano con una técnica de detección del ADN de aspergillus, como es la PCR de aspergillus, podría reducir la incidencia de AI probable o probada según los criterios...

  17. Ser mulher portadora do HPV: uma abordagem cultural Ser mujer portadora de VPH: enfoque cultural Being a woman with HPV: a cultural approach

    Directory of Open Access Journals (Sweden)

    Leilane Barbosa de Sousa

    2008-12-01

    Full Text Available Este trabalho foi realizado com o objetivo de investigar o nível de conhecimento das mulheres sobre o HPV, para, a partir daí, identificar crenças, mitos e tabus sobre a doença e analisar a influência destes elementos culturais no comportamento da mulher. Realizamos um estudo baseado nos pressupostos da teoria do cuidado transcultural. A pesquisa foi desenvolvida a partir do depoimento de quinze mulheres que realizavam tratamento para HPV. Através da investigação, foi possível perceber que, apesar das inúmeras fontes de informação sobre DST, o HPV ainda é uma doença desconhecida e cercada de mistério. Este desconhecimento, interagindo com fatores culturais, favorece o desenvolvimento de conceitos equivocados, tais como crenças e mitos.Este trabajo fue realizado con el objetivo de investigar el nivel de conocimientos de las mujeres sobre el VPH, para así identificar creencias, mitos y tabúes sobre la enfermedad y analizar la influencia de estos elementos culturales en el comportamiento de la mujer. Realizamos un estudio basado en los supuestos de la teoría del cuidado transcultural. La investigación fue realizada en base a los testimonios de quince mujeres en tratamiento para VPH. A través de la investigación fue posible percibir que, a pesar de las innúmeras fuentes de información sobre ETS, el VPH es aún una enfermedad desconocida y rodeada de misterios. Esta falta de conocimiento asociados a los factores culturales, favorecen el desarrollo de conceptos equivocados, tales como creencias y mitos.This study was carried out in order to investigate the level of women's knowledge about HPV, to identify beliefs, myths and taboos about HPV, as well as to analyze the influence of these cultural elements on women's behavior. We performed a study based on the transcultural care theory. The research was developed from the testimonies of 15 women who were undergoing treatment for HPV. Through the investigation, it was possible to

  18. HPV Detection and genotyping in males from the city of Córdoba, Argentina Detección y genotipificación de VPH en varones de la ciudad de Córdoba, Argentina

    Directory of Open Access Journals (Sweden)

    F. Venezuela

    2010-09-01

    Full Text Available A wide range of human papillomavirus (HPV types can infect the anogenital region of males. Although there is a vast knowledge on HPV infections in women as well as on their association with cervical cancer, the study of HPV infections in males is scarce and controversial. The aim of the present work was to detect and typify HPV infections of the anogenital region in males and analyze the associated risk factors in the population studied. Anogenital samples from 37 patients (30 of whom were HIV carriers attending the Infectology Service at the Hospital Nacional de Clínicas in Córdoba, Argentina, were studied. Nine of these patients tested HPV-positive and five out of these nine were found to have mixed infections, being 18 and 61 the most frequent genotypes. There was a significant correlation between the HPV-positive patients and those having an HPV-compatible lesion or AIDS. The present work is the first study in the city of Cordoba which contributes relevant results to the knowledge of HPV infection and to the possible implementation of measures for its prevention.Un amplio espectro de tipos de virus papiloma humano (VPH puede infectar la zona anogenital de los varones. Si bien existe un vasto conocimiento de la infección por VPH en las mujeres y su asociación con el cáncer de cérvix, el estudio de la infección por VPH en los varones ha sido escaso y sus resultados controvertidos. El presente trabajo tuvo como objetivo detectar y tipificar infecciones por VPH en la región anogenital de varones y analizar los factores de riesgo asociados en la población estudiada. Se estudiaron muestras anogenitales de 37 pacientes (30 portadores del VIH que asistieron al Servicio de Infectología del Hospital Nacional de Clínicas de la ciudad de Córdoba. Nueve resultaron positivas para VPH, de las que 5 correspondían a infecciones mixtas. Los genotipos de mayor frecuencia fueron el 18 y el 61. Hubo una correlación significativa entre los

  19. Evaluación del Impacto de las Estrategias de Enseñanza Aprendizaje para Concienciar a los adolescentes acerca del Virus De Papiloma Humano (VPH).

    OpenAIRE

    Hernández, Fabián; Pérez, Wendy; Arias Lara, Sergio Alejandro

    2015-01-01

    El Virus de Papiloma Humano (VPH) es una infección de transmisión sexual (ITS) cuya propagación se ha acelerado en los últimos 20 años en Venezuela y ocupa actualmente un lugar importante entre las estadísticas de ITS más padecidas por adolescentes en el Táchira. La investigación tuvo como objetivo la evaluación del impacto al aplicar las estrategias de enseñanza y aprendizaje para concienciar a los adolescentes del Tercer año Sección “C” del Colegio “Santa Bárbara”, Municipio San Cristóbal e...

  20. Formative research to shape HPV vaccine introduction strategies in Peru Investigación formativa relacionada con el diseño de estrategias para introducir la vacuna contra el VPH en Perú

    Directory of Open Access Journals (Sweden)

    Rosario M Bartolini

    2010-06-01

    Full Text Available OBJECTIVE: To understand the sociocultural environment, health systems' capacities, and policy processes related to cervical cancer and HPV vaccines in order to inform HPV vaccine introduction. MATERIAL AND METHODS: Mixed-method formative research using qualitative and quantitative data collection techniques. Participants included girls, parents, community leaders, health and education officials, and policymakers. RESULTS: Respondents, including policymakers, generally supported HPV vaccine introduction, due partly to appreciation for the benefits of vaccination and the desire to prevent cancer. Community-level concerns regarding safety and quality of services will need to be addressed. The immunization system in Peru is strong and has capacity for including the HPV vaccine. CONCLUSION: Formative research provides key insights to help shape an effective program for HPV vaccine introduction.OBJETIVO: Comprender el contexto sociocultural, las capacidades del sistema de salud y las condiciones políticas vinculadas al cáncer cervical y a la vacuna contra el VPH para diseñar una estrategia apropiada de introducción de la vacuna contra el VPH. MATERIAL Y MÉTODOS: Investigación formativa usando técnicas cualitativas y cuantitativas. Los participantes incluyeron niños, padres, líderes, funcionarios del sector salud y educación, y diseñadores de políticas. RESULTADOS: Generalmente se apoya la introducción de la vacuna contra el VPH, dado que se aprecian los beneficios de la vacunación y se desea prevenir el cáncer. En la comunidad se encontraron preocupaciones sobre seguridad, confianza y calidad de atención. El sistema de inmunizaciones en el Perú es eficiente y tiene la capacidad para incluir la vacuna contra el VPH. CONCLUSIONES: La investigación formativa permite comprender elementos clave que ayudan a diseñar un programa efectivo para la introducción de la vacuna contra el VPH.

  1. La baja utilidad de la determinación del ADN del VPH en la región distal de la uretra masculina Low usefulness of HPV DNA determination in the distal region of the male urethra

    Directory of Open Access Journals (Sweden)

    Ahideé G Leyva-López

    2003-01-01

    Full Text Available OBJETIVO: Determinar la prevalencia uretral del ácido desoxirribonucleico del virus de papiloma humano, condilomatosis clínica y subclínica, en hombres cuyas parejas sexuales tuvieron el antecedente de neoplasia intraepitelial cervical. MATERIAL Y MÉTODOS: De octubre de 1997 a agosto de 1998 se hizo un estudio transversal; se incluyeron 200 hombres de entre 17 a 64 años de edad, referidos a la Coordinación de Oncología del Instituto Nacional de Perinatología, de la Ciudad de México, porque sus parejas regulares sexuales tuvieron el antecedente de neoplasia intraepitelial cervical. Se llevó a cabo un examen físico del pene (penoscopía con la aplicación de ácido acético a 3-5%, y con el uso de un colposcopio se localizaron y evaluaron zonas acetoblancas y cambios vasculares, interpretados como anormales, asociados con la infección por el virus del papiloma humano. La determinación del ADN de VPH se verificó por PCR e hibridación en línea reversa. El análisis estadístico exploratorio y univariante se realizó con el paquete Stata V6.0. RESULTADOS: En las 200 muestras recolectadas de células exfoliadas de la uretra el gen de beta-globina estuvo presente en 93.5% (187/200, y el ácido desoxirribonucleico del virus del papiloma humano fue detectable solamente en 2% (4/187 de los sujetos. Por medio de la penoscopía se observó la presencia de zonas acetoblancas en 43% (81/187 de los sujetos. CONCLUSIONES: En este estudio se observa que la presencia del ácido desoxirribonucleico del virus del papiloma humano en la uretra masculina es poco común, como lo reportan estudios internacionales. Es necesario realizar investigaciones que evalúen esta presencia en glande y surco balano prepucial, en comparación con la región distal de la uretra.OBJECTIVE: To assess the prevalence of Human Papillomavirus (HPV Deoxyribonucleic acid (DNA, and of clinical and subclinical condilomatosis in men whose sex partners had been diagnosed with

  2. Eliminating PCR contamination

    International Nuclear Information System (INIS)

    Fox, J.C.; Ait-Khaled, Mounir; Webster, Alison; Emery, V.C.

    1991-01-01

    The sensitivity of polymerase chain reaction (PCR) can mean that even very low levels of contamination with the target DNA will result in a positive signal. At present this aspect is a major limitation in the use of PCR as a routine diagnostic method. By exposing PCR reagents to UV light, contaminating DNA can be inactivated, thus providing an opportunity to eradicate false positive reactions. UV irradiation was applied to PCR systems used for detection of human cytomegalovirus CMV and human immunodeficiency virus (HIV) and shown to be effective in eradicating both laboratory encountered contamination and plasmid DNA (below 100 pg) added to PCR systems prior to UV exposure. Sensitivity of a PCR system to amplify the long terminal repeat (LTR) sequence of HIV-1 was not affected by the irradiation procedure; however, ultimate sensitivity of a PCR system for the amplification of an early gene pro-motor sequence of the CMV genome was reduced 1000-fold. UV irradiation did not affect the size of the PCR product as determined by strand separating polyacrylamide gel electrophoresis of a 32 P-labelled amplimer. Thus, a simple pre-exposure to UV light would seem a worth-wile step to incorporate into PCR protocols provided that the effects on sensitivity have been determined empirically for each PCR system. (author). 11 refs.; 3 figs

  3. La perspectiva de los agentes sanitarios sobre la incorporación programática de la autotoma del test de VPH

    Directory of Open Access Journals (Sweden)

    Mariana Curotto

    Full Text Available Resumen: El objetivo de este estudio fue analizar la percepción que poseen los agentes sanitarios sobre el ofrecimiento de la autotoma del test de VPH a las mujeres y el grado de acuerdo de los agentes para incorporarla a sus tareas diarias. Para ello, se aplicó una encuesta auto-administrada a 127/191 agentes sanitarios que participaron del Proyecto EMA (Proyecto Evaluación Modalidad Autotoma, llevado a cabo en la provincia de Jujuy (Argentina entre 2012-2013. Los agentes sanitarios que tuvieron y no la experiencia de ofrecer la autotoma manifestaron un alto grado de acuerdo para la adopción de la estrategia (78,7%, dado su potencial para prevenir el cáncer cervicouterino y los aportes que brinda al cuidado de la salud de las mujeres bajo su cobertura. Sin embargo, señalaron la sobrecarga de trabajo y los problemas de articulación con el sistema formal de salud, como los principales obstáculos para ofrecer esta modalidad en el futuro. Este estudio encontró que la autotoma es una práctica que puede ser adoptada por los agentes sanitarios de la provincia de Jujuy, pero debe ir acompañada de acciones de apoyo por parte del sistema de salud formal.

  4. Redes sociales de apoyo y género: vivencia de mujeres con VPH, displasias y cáncer cervicouterino

    Directory of Open Access Journals (Sweden)

    María del Carmen Castro Vásquez

    2014-01-01

    Full Text Available El objetivo del artículo es analizar el funcionamiento y el tipo de redes sociales de apoyo de mujeres diagnosticadas con infección por el virus del papiloma humano ( VPH , displasias y cáncer cervical in situ (Cacu, diferenciando la vivencia por tipo de diagnóstico. Se trata de un estudio cualitativo: se realizaron 34 entrevistas semiestructuradas a mujeres en dos clínicas de displasias en Hermosillo, Sonora. Encontramos que cada diagnóstico tiene connotaciones distintas en la percepción individual y en el imaginario social del grupo de pertenencia, que lleva a las mujeres a una vivencia cargada de incertidumbre y angustia; es en base a estas connotaciones resultantes de las desigualdades de género que las parti- cipantes diseñan sus gestiones de apoyo para resolver la atención médica y organizar su vida familiar y doméstica.

  5. Conductas sexuales de riesgo y actividades preventivas frente al cáncer de cuello uterino en mujeres universitarias vacunadas frente al VPH

    Directory of Open Access Journals (Sweden)

    Ana Fernández-Feito

    2018-05-01

    Full Text Available Resumen: Objetivo: Estimar la asociación entre la vacunación frente al virus papiloma humano (VPH y las conductas sexuales de riesgo, así como la participación en el Programa de Detección Precoz de Cáncer de Cuello Uterino (PDPCCU. Diseño: Estudio descriptivo transversal. Emplazamiento: Facultad de Medicina y Ciencias de la Salud, Facultad de Derecho y Facultad de Economía y Empresa (Universidad de Oviedo. Participantes: Estudiantes universitarias. Mediciones principales: Se recogió información sobre métodos anticonceptivos, conducta sexual, conocimientos sobre VPH y participación en el PDPCCU. Se estimaron proporciones y odds ratio (OR con sus correspondientes intervalos de confianza al 95% (IC 95%. Resultados: El 67,7% de la muestra estaban vacunadas frente al VPH. Un total de 216 mujeres (65,3% eran sexualmente activas. El 67,6% utilizaba un método de barrera en la relación actual, siendo menos frecuente entre las mujeres no vacunadas (54,9% frente al 75,4% en estudiantes vacunadas (p = 0,002. El riesgo de mantener al menos una conducta sexual de riesgo era mayor entre las mujeres no vacunadas: OR 2,29 (IC 95%: 1,29-4,07. La probabilidad de realizar una citología dentro del PDPCCU fue mayor entre las mujeres no vacunadas: OR 2,18 (IC 95%: 1,07-4,47. Conclusiones: La prevalencia de conductas sexuales de riesgo en mujeres no vacunadas es elevada y se relaciona con la no utilización de métodos de barrera. La vacunación frente al VPH puede influir en la conducta sexual y en la participación en PDPCCU. Se debería reforzar la información que reciben los jóvenes sobre anticoncepción, enfermedades de transmisión sexual y prevención del cáncer. Abstract: Aim: To estimate the association between the human papillomavirus (HPV vaccine and sexual risk behaviour, as well as the participation in the Cervical Cancer Screening Program (CCSP. Design: Cross-sectional study. Location: School of Medicine and Health Sciences

  6. External PCR, ASN's decision

    International Nuclear Information System (INIS)

    Anon.

    2012-01-01

    The French law imposes in some situations the presence of a person skilled in radiation protection (PCR). This article describes the cases when this person must belong to the staff of the enterprise or when this person may be sub-contracted. For instance in most nuclear facilities the PCR must be on the payroll, for enterprises dedicated to nuclear transport the PCR's job can be sub-contracted. A decision given by the ASN (French Nuclear Safety Authority) sets the minimal requests (in terms of training, job contract, activities) of the sub-contracted PCR. (A.C.)

  7. Sensado de variables mediante terminal Android

    OpenAIRE

    Altaba Rosas, Mar

    2017-01-01

    El presente documento describe los procesos de diseño y desarrollo de un sistema que, a través de una aplicación móvil, sirve como dispositivo para el registro de la actividad cardíaca del paciente, mediante la obtención del electrocardiograma (ECG), y que permite detectar irregularidades para posteriormente, en caso que fuera necesario, poder enviar los datos adquiridos al profesional sanitario pertinente para que éste los analice. El sistema tiene dos componentes diferenciados, por un lado,...

  8. Reconciliando modularidad y eficiencia mediante atajos

    OpenAIRE

    Marco Gómez, Jordi; Franch Gutiérrez, Javier

    1997-01-01

    Se presenta en este artículo una propuesta para el desarrollo de programas eficientes en el marco de la programación con tipos abstractos de datos (TAD), con el objetivo de respetar la estructura modular de los programas propia de este ámbito. La propuesta se centra en el concepto de atajo como camino eficiente de acceso a los datos, alternativo al acceso mediante las operaciones propias del TAD, y se desarrolla sobre un TAD concreto, el almacén de elementos. La definición de los atajos es al...

  9. Inverse fusion PCR cloning.

    Directory of Open Access Journals (Sweden)

    Markus Spiliotis

    Full Text Available Inverse fusion PCR cloning (IFPC is an easy, PCR based three-step cloning method that allows the seamless and directional insertion of PCR products into virtually all plasmids, this with a free choice of the insertion site. The PCR-derived inserts contain a vector-complementary 5'-end that allows a fusion with the vector by an overlap extension PCR, and the resulting amplified insert-vector fusions are then circularized by ligation prior transformation. A minimal amount of starting material is needed and experimental steps are reduced. Untreated circular plasmid, or alternatively bacteria containing the plasmid, can be used as templates for the insertion, and clean-up of the insert fragment is not urgently required. The whole cloning procedure can be performed within a minimal hands-on time and results in the generation of hundreds to ten-thousands of positive colonies, with a minimal background.

  10. Molecular diagnostic PCR handbook

    International Nuclear Information System (INIS)

    Viljoen, G.J.; Crowther, J.R.; Nel, L.H.

    2005-01-01

    The uses of nucleic acid-directed methods have increased significantly in the past five years and have made important contributions to disease control country programmes for improving national and international trade. These developments include the more routine use of PCR as a diagnostic tool in veterinary diagnostic laboratories. However, there are many problems associated with the transfer and particularly, the application of this technology. These include lack of consideration of: the establishment of quality-assured procedures, the required set-up of the laboratory and the proper training of staff. This can lead to a situation where results are not assured. This book gives a comprehensive account of the practical aspects of PCR and strong consideration is given to ensure its optimal use in a laboratory environment. This includes the setting-up of a PCR laboratory; Good Laboratory Practice and standardised PCR protocols to detect animal disease pathogens. Examples of Standard Operating Procedures as used in individual specialist laboratories and an outline of training materials necessary for PCR technology transfer are presented. The difficulties, advantages and disadvantages in PCR applications are explained and placed in context with other test systems. Emphasis is placed on the use of PCR for detection of pathogens, with a particular focus on diagnosticians and scientists from the developing world. It is hoped that this book will enable readers from various disciplines and levels of expertise to better judge the merits of PCR and to increase their skills and knowledge in order to assist in a more logical, efficient and assured use of this technology

  11. Análisis de la prevalencia de tipos de VPH en muestras de citologías ginecológicas en la población no vacunada. Estudio en la Comunidad de Extremadura

    OpenAIRE

    Fernández de Mera, José Juan

    2016-01-01

    Con la intención de conocer la situación de la infección por VPH entre las mujeres extremeñas no vacunadas contra el virus, se analizan las 355.938 citologías estudiadas en la Comunidad de Extremadura, bajo un plan de cribado oportunista, entre los años 2002 y 2013. Éstas corresponde a 173.841 mujeres, de entre las que se extrae una muestra sin lesiones citológicas y sin antecedentes ginecológicos de 593 mujeres. Los diagnósticos citológicos se clasificaron atendiendo al consenso de Bethesda ...

  12. Tipificación del vph en cáncer de cuello uterino en la población venezolana

    OpenAIRE

    Suárez, Carmen María; Mijares Briñez, Alirio; Castillo Marrero, Livia; Briceño, Josefa María

    2006-01-01

    OBJETIVOS: Realizar la tipificación del virus del papiloma humano en pacientes con cáncer de cuello uterino en la población venezolana. MÉTODOS: Se incluyeron 53 pacientes con diagnóstico de cáncer de cuello uterino entre abril de 2004 y noviembre de 2005. Se realizó tipificación del virus del papiloma humano mediante la reacción en cadena de la polimerasa para los tipos 6, 11, 16, 18, 31, 33 y 35. Se analizaron otras variables como edad, estadio y tipo histológico. RESULTADOS: La edad promed...

  13. Detección molecular de VPH por PCR-RFLP en muestras del área ano-genital de hombres en el Estado Mérida, Venezuela

    OpenAIRE

    Quintero, Militza; Cruz, Jhon; Bastidas, Marco; Hernández, Danmarys; Rodríguez, Adriana; Puig, Juan

    2013-01-01

    Objetivo: Determinar el porcentaje de positividad y tipo de virus de papiloma humano en el área genitoanal de hombres, relación con la edad, sitio anatómico y tipo de muestra, en el Departamento de Biología, Laboratorio de Biología y Medicina Experimental Facultad de Ciencias. LABIOMEX Universidad de Los Andes. Métodos: Estudio descriptivo, transversal, de 882 muestras del área genito-anal de hombres que acudieron voluntariamente a la consulta para detección y tipificación de virus de papilom...

  14. PCR in forensic genetics

    DEFF Research Database (Denmark)

    Morling, Niels

    2009-01-01

    Since the introduction in the mid-1980s of analyses of minisatellites for DNA analyses, a revolution has taken place in forensic genetics. The subsequent invention of the PCR made it possible to develop forensic genetics tools that allow both very informative routine investigations and still more...... and more advanced, special investigations in cases concerning crime, paternity, relationship, disaster victim identification etc. The present review gives an update on the use of DNA investigations in forensic genetics.......Since the introduction in the mid-1980s of analyses of minisatellites for DNA analyses, a revolution has taken place in forensic genetics. The subsequent invention of the PCR made it possible to develop forensic genetics tools that allow both very informative routine investigations and still more...

  15. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi

    Directory of Open Access Journals (Sweden)

    Danielle C. Leal

    2011-07-01

    Full Text Available Conventional PCR (PCRTeq for diagnosing Theileria equi and multiplex PCR (M/PCRTeq-Bc for diagnosing T. equi and Babesia caballi were comparatively evaluated with nested PCR (N/PCR-Teq for diagnosing equine piroplasmosis. In DNA sensitivity determinations, in multiple dilutions of equine blood that had tested positive for T. equi, PCR-Teq and N/PCR-Teq detected hemoparasite DNA in the larger dilutions (1:128, but did not differ significantly from the M/PCRTeq-Bc (1:64. In analyses on equine serum tested by ELISA, there was high agreement between this serological test and PCR-Teq (k = 0.780 and moderate agreement with N/PCR-Teq (k = 0.562 and M/PCRTeq-Bc (k = 0.488. PCR-Teq found a higher frequency of T. equi both in extensively and intensively reared horses, but this was not significant in relation to N/PCR-Teq (P>0.05, and both PCRs indicated that there was an endemic situation regarding T. equi in the population of horses of this sample. PCR-Teq was only significantly different from M/PCR-Teq-Bc (P<0.05. PCR-Teq presented high sensitivity and specificity, comparable to N/PCR-Teq, but with the advantage of higher speed in obtaining results and lower costs and risks of laboratory contamination. This accredits PCR-Teq for epidemiological studies and for determinations on affected horses.

  16. Acceso a la información de mujeres con VPH, displasia y cáncer cervical in situ Access to information by women with HPV, cervical dysplasia and cancer in situ

    Directory of Open Access Journals (Sweden)

    Ma del Carmen Castro-Vásquez

    2010-06-01

    Full Text Available OBJETIVO: Presentar un análisis relacional de cómo mujeres diagnosticadas con el virus del papiloma humano (VPH, displasia del cuello del útero o neoplasias del cuello uterino, reciben y/o acceden a la información y cómo la viven en sus relaciones cercanas. MATERIAL Y MÉTODOS: En 2008 se realizaron 34 entrevistas cualitativas a mujeres en dos clínicas de colposcopía de la Secretaría de Salud, en Hermosillo, Sonora. El análisis se basó en la teoría fundamentada. RESULTADOS: Existe una franca analogía entre cáncer cervicouterino (CaCu y muerte, una amplia desinformación sobre VPH y displasias y una práctica persistente entre los médicos de no ofrecer información oportuna y clara a las pacientes. Existe una apreciación estigmatizante hacia la infección por VPH que afecta las relaciones cercanas de las mujeres. CONCLUSIÓN: A pesar de la necesidad de las pacientes de obtener información, no la exigen al médico, lo que contribuye a su desconfianza y angustia.OBJECTIVE: To present a relational analysis of how women who are diagnosed with the human papilloma virus (HPV, cervical dysplasia or cervical neoplasia receive or seek information, and how they experience this process within their immediate relationships. MATERIALS AND METHODS: In 2008, 34 qualitative interviews were carried out with women at two Secretary of Health colposcopy clinics in Hermosillo, Sonora. Analysis was based on grounded theory. RESULTS: There is a patent analogy between cervical cancer and death, much disinformation about HPV and dysplasias, and a persistent lack of timely and clear information given to patients by doctors. There is a stigma attached to HPV infection which affects women's immediate relationships. CONCLUSION: Despite patients' need to obtain information, they do not demand it from their doctor, which contributes to their anguish and distrust.

  17. Por qué la vacuna contra el VPH es importante para mi familia: historia de una sobreviviente de cáncer de cuello uterino (Why HPV Vaccine is Important to My Family: The Story of a Cervical Cancer Survivor)

    Centers for Disease Control (CDC) Podcasts

    A una mamá joven se le vino el mundo encima cuando le diagnosticaron cáncer de cuello uterino. Entérese de lo que está haciendo para proteger a sus hijos de los cánceres asociados al VPH.

  18. Distinción de especies del género Persea mediante RAPD e ISSR de ADN

    OpenAIRE

    Reyes-Alemán, Juan Carlos; Valadez-Moctezuma, Ernestina; Simuta-Velázco, Lisandro; Barrientos-Priego, Alejandro Facundo; Gallegos-Vázquez, Clemente

    2013-01-01

    Con la finalidad de establecer bases para diferenciar parte de la diversidad genética de Persea y en especial del subgénero Persea resguardado en la colección nacional de germoplasma de aguacate de México, se estudiaron ocho especies (P. americana, P. steyermarkii, P. schiedeana, P. lingue, P. nubigena, P. floccosa, P. cinerascens y P. indica) con marcadores moleculares mediante las técnicas de RAPD e ISSR, donde los productos de PCR fueron separados en geles de acrilamida. Las huellas de ADN...

  19. PCR, exit stage left ...

    CERN Multimedia

    2004-01-01

    The Prevessin Control Room during LEP's start up in 1989. The Prévessin Control Room (PCR) was recently engulfed in a wave of nostalgia. The PCR, scene of some of the greatest moments in CERN's history, is being dismantled to prepare for a complete overhaul. In February 2006, a new combined control centre for all the accelerators will open its doors on the same site, together with a new building currently under construction (see Bulletin issue 27/2004 of 28 June 2004). This marks the end of an important chapter in CERN's history. The Prévessin Control Room saw its first momentous event 28 years ago when the 400 GeV beam for the SPS was commissioned in the presence of Project Leader John Adams. It was also here that the first proton-antiproton collisions were observed, in 1981. Eight years later, in 1989, operators and directors alike jumped for joy at the announcement of the first electron-positron collisions at the start up of LEP, the biggest accelerator in the world. Today the 80 terminals and PCs have b...

  20. Quantitative (real-time) PCR

    International Nuclear Information System (INIS)

    Denman, S.E.; McSweeney, C.S.

    2005-01-01

    Many nucleic acid-based probe and PCR assays have been developed for the detection tracking of specific microbes within the rumen ecosystem. Conventional PCR assays detect PCR products at the end stage of each PCR reaction, where exponential amplification is no longer being achieved. This approach can result in different end product (amplicon) quantities being generated. In contrast, using quantitative, or real-time PCR, quantification of the amplicon is performed not at the end of the reaction, but rather during exponential amplification, where theoretically each cycle will result in a doubling of product being created. For real-time PCR, the cycle at which fluorescence is deemed to be detectable above the background during the exponential phase is termed the cycle threshold (Ct). The Ct values obtained are then used for quantitation, which will be discussed later

  1. One-stop polymerase chain reaction (PCR): An improved PCR ...

    African Journals Online (AJOL)

    Yomi

    2011-12-21

    Dec 21, 2011 ... membrane filtration was carried out with a commercial PCR product purification kit (Generay, Shanghai), according to the manufacture's instruction. In brief, 50 µl PCR product was mixed thoroughly with binding buffer, and the resultant mixture was loaded directly onto a silica membrane Gelclean column.

  2. Por qué la vacuna contra el VPH es importante para mi familia: historia de una sobreviviente de cáncer de cuello uterino (Why HPV Vaccine is Important to My Family: The Story of a Cervical Cancer Survivor)

    Centers for Disease Control (CDC) Podcasts

    2013-05-06

    A una mamá joven se le vino el mundo encima cuando le diagnosticaron cáncer de cuello uterino. Entérese de lo que está haciendo para proteger a sus hijos de los cánceres asociados al VPH.  Created: 5/6/2013 by National Center for Immunization and Respiratory Diseases (NCIRD).   Date Released: 10/30/2013.

  3. Digital PCR: A brief history

    OpenAIRE

    Morley, Alexander A.

    2014-01-01

    Digital PCR for quantification of a target of interest has been independently developed several times, being described in 1990 and 1991 using the term “limiting dilution PCR” and in 1999 using the term “digital PCR”. It came into use in the decade following its first development but its use was cut short by the description of real-time PCR in 1996. However digital PCR has now had a renaissance due to the recent development of new instruments and chemistry which have made it a much simpler and...

  4. Detección molecular de las translocaciones más comunes en Leucemia aguda mediante RT-PCR

    Directory of Open Access Journals (Sweden)

    L. García

    2001-07-01

    Full Text Available Evaluar la incidencia de las translocaciones t(4;11, t(1;19, t(9;22 y t(12;21 en leucemia linfoide aguda (LLA y t(15;17, t(8;21 e Inv.(16 en leucemia mieloide aguda (LMA. Correlacionar los resultados obtenidos con el diagnóstico morfológico y citogenético.

  5. Cervical cancer and the HPV link: identifying areas for education in Mexico City's public hospitals El cáncer cervicouterino y su relación con el VPH: identificación de temas de actualización en hospitales públicos de la Ciudad de México

    Directory of Open Access Journals (Sweden)

    Tess Aldrich

    2006-06-01

    Full Text Available OBJECTIVE:To assess Mexico City physicians' knowledge and practices regarding cervical cancer and human papillomavirus (HPV to compare obstetricians/gynecologists (ob/gyns and general practitioners (GPs on these variables MATERIAL AND METHODS: In April 2003, 187 ob/gyns and GPs working in 15 hospitals affiliated with the Federal District Secretary of Health (SSDF completed a self-administered questionnaire. Pearson's chi-square tests were used to compare ob/ gyns and GPs on outcome variables RESULTS: Nearly all providers (93% identified HPV as the principal cause of cervical cancer. Ob/gyns had more detailed knowledge about HPV than GPs and were more likely to have heard of common oncogenic strains (p=.000. Sixteen percent of all physicians incorrectly stated that Pap tests should be performed every six months regardless of previous results, and 17% recommended hysterectomy as an option for treating mild or moderate dysplasia CONCLUSIONS: While SSDF physicians had basic knowledge about the cervical cancer-HPV link, screening and management norms are priority areas for educational interventions.OBJETIVO: Evaluar el conocimiento y las prácticas de los proveedores de servicios de salud en la Ciudad de México sobre el cáncer cervicouterino y el virus del papiloma humano (VPH; comparar a este respecto a ginecoobstetras (GO y médicos generales (MG MATERIAL Y MÉTODOS: En abril del 2003, 187 GO y MG empleados en 15 hospitales afiliados a la Secretaría de Salud del Distrito Federal (SSDF completaron un cuestionario autoaplicado. Se utilizó la prueba de ji cuadrada de Pearson para evaluar las diferencias entre GO y MG RESULTADOS: Casi todos los participantes (93% identificaron el VPH como la causa principal del cáncer cervicouterino. Los GO mostraron un conocimiento más detallado del VPH que los MG, con más probabilidad de haber escuchado de las cepas oncogénicas comunes del VPH (p= 0.000. Un 16% de los médicos contestó incorrectamente que

  6. Análisis taxonómico y funcional del microbioma humano mediante aproximaciones clásicas, moleculares y metagenómicas

    OpenAIRE

    Cabrera Rubio, Raúl

    2014-01-01

    La presente tesis muestra distintas aproximaciones para el estudio del microbioma humano. Éstas han ido desde la secuenciación masiva de productos de PCR, la pirosecuenciación directa del ADN ambiental, la elaboración de librerías de fósmidos y por último el aislamiento de especies presentes en el microbioma mediante sembrado de la muestra. Todas estas técnicas tienen sus ventajas y desventajas, pero todas ellas son complementarias para el estudio de un determinado microbioma. Además la elabo...

  7. Methylation-Specific PCR Unraveled

    Directory of Open Access Journals (Sweden)

    Sarah Derks

    2004-01-01

    Full Text Available Methylation‐specific PCR (MSP is a simple, quick and cost‐effective method to analyze the DNA methylation status of virtually any group of CpG sites within a CpG island. The technique comprises two parts: (1 sodium bisulfite conversion of unmethylated cytosine's to uracil under conditions whereby methylated cytosines remains unchanged and (2 detection of the bisulfite induced sequence differences by PCR using specific primer sets for both unmethylated and methylated DNA. This review discusses the critical parameters of MSP and presents an overview of the available MSP variants and the (clinical applications.

  8. Design and methods of the evaluation of an HPV-based cervical cancer screening strategy in Mexico: the Morelos HPV study Diseño y métodos de la evaluación del uso de la prueba de virus de papiloma humano para tamizaje de cáncer cervical en México: el estudio de VPH en Morelos

    Directory of Open Access Journals (Sweden)

    Yvonne Flores

    2002-07-01

    enrollment activities of the Morelos HPV study are the basis for a prevalent case-control study and a prospective cohort study that will investigate the natural history of HPV infections and determine if an HPV-based screening strategy is a safe and cost-effective alternative to Pap screening.Objetivo. Describir los métodos y el diseño del Estudio de VPH en Morelos. El objetivo principal de este estudio es examinar el uso de dos diferentes técnicas para obtener muestras de ADN de VPH, autotoma vaginal y toma clínica cervical, para detectar lesiones cervicales preinvasoras y cáncer. Material y métodos. Este estudio se realizó en el marco del Programa de Detección Oportuna de Cáncer Cervical (DOC del Instituto Mexicano del Seguro Social (IMSS en Morelos. Un total de 7 868 mujeres aceptaron participar en el estudio durante los meses de mayo a octubre de 1999. Esta muestra es representativa de la población de mujeres que acudieron a los programas de DOC en las 23 clínicas del IMSS en Morelos durante ese año. Se les proporcionó una explicación detallada del estudio a las participantes antes de que firmaran una carta de consentimiento informado. Se obtuvo información básica de todas las participantes y se usó el formato oficial de registro de datos del programa institucional de DOC del IMSS. Además se seleccionó una submuestra aleatoria de 1 069 participantes que fueron entrevistadas durante su visita inicial para obtener información adicional sobre los factores de riesgo de cáncer cervical, aceptabilidad de las pruebas de Papanicolaou (Pap y VPH-autotoma, y los costos de las pacientes. Todas las participantes se tomaron una muestra de exudado vaginal para la prueba de VPH-autotoma. Posteriormente, se les realizó una exploración pélvica para obtener muestras cervicales para las pruebas de VPH-clínica y Pap. Se evaluó la información de 7 732 mujeres con resultados completos de las tres pruebas. A las 1 147 mujeres que recibieron un diagnóstico positivo

  9. pcr

    African Journals Online (AJOL)

    DR. AMINU

    Keywords: Polymerase chain reaction, Diagnosis, Bacteria, Infections. INTRODUCTION ... used to amplify a piece of DNA by in-vitro enzymatic replication (David and .... receiving antimicrobial treatment, or when the causative agents are small ...

  10. Detección de Mycoplasma genitalium mediante Reacción en Cadena de la Polimerasa en muestras urogenitales de individuos cubanos sexualmente activos

    Directory of Open Access Journals (Sweden)

    Brian Arturo Mondeja-Rodríguez

    2014-04-01

    Full Text Available El diagnóstico de las infecciones por Mycoplasma genitalium mediante métodos bacteriológicos tradicionales resulta laborioso y poco práctico. Es por ello que los métodos moleculares basados en la amplificación del ADN se utilizan con fines diagnósticos de las infecciones causadas por este microorganismo. En Cuba se han realizado pocos estudios sobre la presencia de M. genitalium en el tracto urogenital. El objetivo de la presente investigación fue detectar M. genitalium en individuos cubanos sexualmente activos mediante la implementación de métodos de PCR simple. Se implementaron dos PCR simples para la detección de fragmentos de 427 pb del gen ARN ribosomal 16S y 281 pb del gen de la adhesina celular MgPa de M. genitalium, que se evaluaron en muestras de exudado endocervical provenientes de 300 mujeres con sintomatología urogenital y muestras de orina de 49 hombres asintomáticos sexualmente activos. Se logró un límite de detección de la PCR del ARNr 16S de aproximadamente 5 copias de genoma por reacción, mientras que para la PCR MgPa se logró la amplificación de solo 50 copias de genoma por reacción. El 3% (10/300 de los exudados endocervicales y el 24,5% (12/49 de las muestras de orina de hombres asintomáticos resultaron positivas mediante ambas PCR. El mayor porcentaje de muestras positivas correspondió a las muestras de orina provenientes de hombres asintomáticos, que resultó superior a lo esperado. El presente trabajo permitirá realizar estudios futuros de caracterización genética y antigénica de las cepas de Mycoplasma genitalium circulantes en Cuba, útiles para conformar un inmunógeno vacunal.

  11. Cost-effectiveness of conventional cytology and HPV DNA testing for cervical cancer screening in Colombia Costo-efectividad de la citología y la tamización con pruebas de ADN-VPH para cáncer de cuello uterino en Colombia

    Directory of Open Access Journals (Sweden)

    Oscar Andrés-Gamboa

    2008-08-01

    Full Text Available OBJECTIVE: To assess cost-effectiveness of conventional cytology and HPV DNA testing for cervical-cancer screening in Colombia. MATERIAL AND METHODS: The National Cancer Institute of Colombia (NCIC in 2007 developed a Markov model on the natural history of cervical cancer; no screening, conventional cytology, and HPV DNA testing were compared. Only direct costs were used. Outcomes comprise cervical cancer mortality, years of life saved, and lifetime costs. Discounted incremental cost-effectiveness ratios were estimated and sensitivity analyses were conducted for key parameters. RESULTS: Depending on the screening strategy a 69-81% mortality reduction might be expected. The HPV DNA testing every five years is a cost-effective strategy (Incremental Cost-Effectiveness Ratio (ICER: USD$44/YLS if the cost per test is under USD$31. The effectiveness was sensitive to coverage and primarily to follow-up. CONCLUSIONS: HPV DNA testing is a cost-effective alternative for screening in Colombia. Not only high coverage but high follow-up rates are critical for successful screening programs.OBJETIVO: evaluar el costo-efectividad de la citología convencional y la prueba de ADN-VPH para tamización de cáncer cervical en Colombia. MATERIAL Y MÉTODOS: el Instituto Nacional de Cancerología de Colombia construyó en 2007 un modelo de Markov de historia natural del cáncer cervical. Se comparó "no tamización", citología convencional y prueba de ADN-VPH. Se utilizaron costos directos. Los desenlaces fueron mortalidad, años de vida ganados y costos. Se calcularon razones de costo-efectividad incremental. Se realizaron análisis de sensibilidad para parámetros clave. RESULTADOS: la mortalidad se redujo 69-81% según la estrategia. La tamización con ADN-VPH cada cinco años es costo-efectiva (ICER (Razón de Costo-Efectividad incremental por sus siglas en inglés: 44 dólares por año de vida saludable si los costos por prueba son menores a 31 dólares. La

  12. Epidemiología de la infección y detección de tipos oncogenéticos del VPH por tecnología de captura de híbridos en mujeres sin aparentes factores de riesgo

    OpenAIRE

    Fuente Villarreal, David de la

    2011-01-01

    Introducción: El virus del papiloma humano (VPH) pertenece al grupo de virus con tropismo por los epitelios, infecta predominantemente la piel y las membranas mucosas produciendo proliferaciones epiteliales benignas o papilomas, que bajo ciertas circunstancias (las cuales si bien aún no han sido definidas a satisfacción, se estima que se relacionan con el estado inmunológico de la paciente, su carga genética, la presencia de ciertos receptores, así como la continua infección por diversos tipo...

  13. Conocimientos, actitudes y prácticas sobre virus de papiloma humano (VPH) y cáncer de cuello uterino en mujeres de 30 y más años de edad, de un barrio ribereño de Asunción, (Bañado Sur). 2012

    OpenAIRE

    Malvina Páez B; María I Rodríguez-Riveros; Elena Kasamatsu; Amalia Castro; Elizabeth Orué; Natalia Lampert; Mónica Ruoti; Mónica Sequera; Graciela Giménez; Laura Mendoza; Pamela Mongelós; Adriana Valenzuela; María A Leguizamón S

    2016-01-01

    Introducción: El cáncer de cuello uterino es un problema de salud pública en Paraguay. Objetivo: Determinar conocimientos, actitudes y prácticas sobre virus del papiloma humano (VPH) y cáncer de cuello uterino en mujeres de 12 Unidades de Salud Familiar (USF) de Bañado Sur-Asunción, periodo abril-octubre 2012. Metodología: Estudio descriptivo de corte transversal, utilizando cuestionario estructurado autoadministrado. Resultados: La edad promedio de las encuestadas fue 42 años, la mayoría en ...

  14. Seguimiento de trayectorias tridimensionales de un quadrotor mediante control PVA

    Directory of Open Access Journals (Sweden)

    Silvia Estellés Martínez

    2014-01-01

    Full Text Available Resumen: Este trabajo presenta el modelado de un quadrotor como un sistema multicuerpo llevado a cabo mediante el software Vehicle- Sim, en el que los diferentes componentes del sistema son descritos mediante una estructura paterno-filial señalando las restricciones físicas entre ellos. Los modelos estructural y aerodinámico han sido desarrollados mediante este software, ampliamente utilizado en la simulación del comportamiento dinámico de vehículos.Sobre el modelo resultante se he desarrollado un algoritmo de control basado en la metodologia PVA con la finalidad de obtener un seguimiento de trayectoria mediante acciones de control suaves. Empleando la metodología convencional de control PVA no es posible estabilizar el vehículo en todos los rangos de posicionamiento lateral (y y longitudinal (x. En este artículo los autores muestran como esta limitación en el diseño de una estrategia de control PVA convencional es solventada con una modificación consistente en sustituir los parámetros constantes del PVA clásico por funciones dependientes del desplazamiento.El sistema de control es implementado para adecuarse a los requerimientos de las actuaciones y se diseña sobre la plataforma de simulación multidominio Simulink. Con la finalidad de obtener una importante mejora en la respuesta de posicionamiento, se im- plementa un generador de trayectorias continuas.Una vez que el modelo es desarrollado y el sistema de control implementado, los autores presentan el modelo matemático y los resultados de las simulaciones realizadas. Éstas validan el empleo tanto de la metodología de control PVA aplicada, como de la alimentación de trayectorias predefinidas, no sólo para la posición, sino también para la velocidad y aceleración. Abstract: In this work the authors present the modelling of a quadrotor as a multibody system carried out with the software VehicleSim, in which the different

  15. Lqr Robusto Mediante Incertidumbre Acotada En Los Datos

    Directory of Open Access Journals (Sweden)

    C. Ramos

    2007-07-01

    Full Text Available Resumen: En este trabajo se presenta el sintonizado del Regulador Lineal Cuadrático (LQR mediante la técnica de incertidumbre acotada en los datos o Bounded Data Uncertainties (BDU con el fin de mejorar la robustez del sistema, planteándose como un Min-Max donde se busca la mejor solución en el peor escenario posible. Así se ofrece un nuevo método guiado de ajuste del LQR, considerando los límites de la incertidumbre. La aplicación a sistemas multidimensionales no es trivial, pues presenta la forma de un Two-Point Boundary Value Problem (TPBVP, el cual se resuelve iterativamente. : Técnicas Minimax, Regularización, Método de Control LQR, Robustez, Incertidumbre, Ecuaciones Matriciales de Riccati, Problema de Valor Límite, Sistemas Multidimensionales

  16. Caracterización a impacto de caucho reciclado mediante elementos finitos

    OpenAIRE

    Escribano Castro, Ane

    2015-01-01

    Análisis de caucho reciclado de manera hiperelástica mediante métodos de ajuste de Mínimos Cuadrados con programa MATLAB y Curve fitting mediante ANSYS. Para la parte viscoelástica se usa Algoritmo de Optimicación con MATLAB. Comprobación de resultados y fiabilidad.

  17. Two-temperature PCR for Microfluidics

    KAUST Repository

    Kodzius, Rimantas

    2010-05-01

    Since its invention in 1983, polymerase chain reaction (PCR) has been the method of choice for DNA amplification. Successful PCR depends on the optimization of several parameters, which is a cumbersome task due to the many variables (conditions and compon

  18. Two-temperature PCR for Microfluidics

    KAUST Repository

    Kodzius, Rimantas; Chang, Donald Choy; Sheng, Ping; Wen, Weijia; Wu, Jinbo; Xiao, Kang; Yu, Vivian

    2010-01-01

    Since its invention in 1983, polymerase chain reaction (PCR) has been the method of choice for DNA amplification. Successful PCR depends on the optimization of several parameters, which is a cumbersome task due to the many variables (conditions and compon

  19. Pathogen Causing Disease of Diagnosis PCR Tecnology

    OpenAIRE

    SEVİNDİK, Emre; KIR, A. Çağrı; BAŞKEMER, Kadir; UZUN, Veysel

    2013-01-01

    Polimerase chain reaction (PCR) with which, the development of recombinant DNA tecnology, a technique commonly used in field of moleculer biology and genetic. Duplication of the target DNA is provided with this technique without the need for cloning. Some fungus species, bacteria, viruses constitutent an important group of pathogenicity in human, animals and plants. There are routinely applied types of PCR in the detection of pathogens infections diseases. These Nested- PCR, Real- Time PCR, M...

  20. Principles and technical aspects of PCR amplification

    National Research Council Canada - National Science Library

    Pelt-Verkuil, Elizabeth van; Belkum, Alex van; Hays, John P

    2008-01-01

    ... to illustrate any particularly important concepts or comments. Indeed, all commercial PCR biotechnology companies offer information about their products on internet sites and in online technical manuals. These online resources will be invaluable for any readers requiring more detailed PCR protocols. The authors have provided references for many PCR co...

  1. The PCR revolution: basic technologies and applications

    National Research Council Canada - National Science Library

    Bustin, Stephen A

    2010-01-01

    ... by leading authorities on the many applications of PCR and how this technology has revolutionized their respective areas of interest. This book conveys the ways in which PCR has overcome many obstacles in life science and clinical research and also charts the PCR's development from time-consuming, low throughput, nonquantitative proced...

  2. Distributed Data Management Service for VPH Applications

    NARCIS (Netherlands)

    Koulouzis, S.; Belloum, A.; Bubak, M.; Lamata, P.; Nolte, D.; Vasyunin, D.; de Laat, C.

    2016-01-01

    For many medical applications, it's challenging to access large datasets, which are often hosted across different domains on heterogeneous infrastructures. Homogenizing the infrastructure to simplify data access is unrealistic; therefore, it's important to develop distributed storage that doesn't

  3. Real-Time PCR (qPCR) Primer Design Using Free Online Software

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most…

  4. Detection of adenoviruses in shellfish by means of conventional-PCR, nested-PCR, and integrated cell culture PCR (ICC/PCR).

    Science.gov (United States)

    Rigotto, C; Sincero, T C M; Simões, C M O; Barardi, C R M

    2005-01-01

    We tested three PCR based methodologies to detect adenoviruses associated with cultivated oysters. Conventional-PCR, nested-PCR, and integrated cell culture-PCR (ICC/PCR) were first optimized using oysters seeded with know amounts of Adenovirus serotype 5 (Ad5). The maximum sensitivity for Ad5 detection was determined for each method, and then used to detect natural adenovirus contamination in oysters from three aquiculture farms in Florianopolis, Santa Catarina State, Brazil, over a period of 6 months. The results showed that the nested-PCR was more sensitive (limit of detection: 1.2 PFU/g of tissue) than conventional-PCR and ICC-PCR (limit of detection for both: 1.2 x 10(2)PFU/g of tissue) for detection of Ad5 in oyster extracts. Nested-PCR was able to detect 90% of Ad5 contamination in harvested oyster samples, while conventional-PCR was unable to detect Ad5 in any of the samples. The present work suggests that detection of human adenoviruses can be used as a tool to monitor the presence of human viruses in marine environments where shellfish grow, and that nested-PCR is the method of choice.

  5. DETECCION DE Brucella abortus POR PCR EN MUESTRAS DE SANGRE Y LECHE DE VACUNOS

    Directory of Open Access Journals (Sweden)

    Xiomara Mosquera C

    2008-12-01

    Full Text Available Objetivo. Evaluar el uso de la Reacción en Cadena de la Polimerasa (PCR para la detección de Brucella abortus en muestras de sangre y leche de vacunos. Materiales y métodos. Este estudio de tipo descriptivo fue realizado durante los años 2004 y 2005. Se analizaron 136 animales de tres fincas localizadas en el municipio de Durania, Norte de Santander, Colombia. Se evaluó la presencia de anticuerpos en la leche mediante la prueba del anillo (PAL. Se amplificó el fragmento de 223pb del gen BCSP31. Se emplearon los cebadores B4 y B5 de la región interna de la secuencia del gen BCSP31 (GenBank, número M20404. Resultados. En aquellos animales positivos se obtuvo una muestra de sangre y leche para el análisis por PCR, la sangre no fue analizada por serología. Se evaluaron diferentes métodos de extracción de ADN. Se encontró que un 13.2% (18/136 de las muestras de leche fueron positivas a la PAL. Se analizaron 33 muestras de leche negativas por PAL de las cuales el 30.3% (10/33 resultaron positivas por PCR. Al analizar las muestras de sangre de los animales positivos por PAL el 94.1% (16/17 fueron positivas por PCR, mientras que el 47% (8/17 de las muestras de leche positivas por PAL, fueron positivas por PCR. Conclusiones. Se demostró la amplificación de un fragmento de ADN de Brucella abortus en muestras de sangre y leche de vacunos. Los resultados preliminares demostraron que es posible usar PCR como prueba diagnóstica de brucelosis en Colombia.

  6. Procedimiento de estabilizacion de mercurio liquido mediante cemento polimerico de azufre,via sulfuro de mercurio

    OpenAIRE

    López Gómez, Félix Antonio; López-Delgado, Aurora; Alguacil, Francisco José; Alonso Gámez, Manuel

    2009-01-01

    Procedimiento para la estabilización de mercurio líquido mediante la obtención de cementos poliméricos de azufre que comprende: (a) transformación del mercurio líquido en sulfuro de mercurio (metacinabrio) mediante reacción química, en condiciones estequiométricas, entre el mercurio y el azufre elemental; y (b) obtención de cemento polimérico de azufre mediante la incorporación el sulfuro de mercurio obtenido en la etapa anterior, en una mezcla estable constituida por áridos, azufre elemental...

  7. Application of reverse transcription-PCR and real-time PCR in nanotoxicity research.

    Science.gov (United States)

    Mo, Yiqun; Wan, Rong; Zhang, Qunwei

    2012-01-01

    Reverse transcription-polymerase chain reaction (RT-PCR) is a relatively simple and inexpensive technique to determine the expression level of target genes and is widely used in biomedical science research including nanotoxicology studies for semiquantitative analysis. Real-time PCR allows for the detection of PCR amplification in the exponential growth phase of the reaction and is much more quantitative than traditional RT-PCR. Although a number of kits and reagents for RT-PCR and real-time PCR are commercially available, the basic principles are the same. Here, we describe the procedures for total RNA isolation by using TRI Reagent, for reverse transcription (RT) by M-MLV reverse transcriptase, and for PCR by GoTaq(®) DNA Polymerase. And real-time PCR will be performed on an iQ5 multicolor real-time PCR detection system by using iQ™ SYBR Green Supermix.

  8. Pitfalls in PCR troubleshooting: Expect the unexpected?

    Directory of Open Access Journals (Sweden)

    Livia Schrick

    2016-01-01

    Full Text Available PCR is a well-understood and established laboratory technique often used in molecular diagnostics. Huge experience has been accumulated over the last years regarding the design of PCR assays and their set-up, including in-depth troubleshooting to obtain the optimal PCR assay for each purpose. Here we report a PCR troubleshooting that came up with a surprising result never observed before. With this report we hope to sensitize the reader to this peculiar problem and to save troubleshooting efforts in similar situations, especially in time-critical and ambitious diagnostic settings.

  9. Absolute quantification by droplet digital PCR versus analog real-time PCR

    Science.gov (United States)

    Hindson, Christopher M; Chevillet, John R; Briggs, Hilary A; Gallichotte, Emily N; Ruf, Ingrid K; Hindson, Benjamin J; Vessella, Robert L; Tewari, Muneesh

    2014-01-01

    Nanoliter-sized droplet technology paired with digital PCR (ddPCR) holds promise for highly precise, absolute nucleic acid quantification. Our comparison of microRNA quantification by ddPCR and real-time PCR revealed greater precision (coefficients of variation decreased by 37–86%) and improved day-to-day reproducibility (by a factor of seven) of ddPCR but with comparable sensitivity. When we applied ddPCR to serum microRNA biomarker analysis, this translated to superior diagnostic performance for identifying individuals with cancer. PMID:23995387

  10. Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology.

    Science.gov (United States)

    Smith, Cindy J; Osborn, A Mark

    2009-01-01

    Quantitative PCR (Q-PCR or real-time PCR) approaches are now widely applied in microbial ecology to quantify the abundance and expression of taxonomic and functional gene markers within the environment. Q-PCR-based analyses combine 'traditional' end-point detection PCR with fluorescent detection technologies to record the accumulation of amplicons in 'real time' during each cycle of the PCR amplification. By detection of amplicons during the early exponential phase of the PCR, this enables the quantification of gene (or transcript) numbers when these are proportional to the starting template concentration. When Q-PCR is coupled with a preceding reverse transcription reaction, it can be used to quantify gene expression (RT-Q-PCR). This review firstly addresses the theoretical and practical implementation of Q-PCR and RT-Q-PCR protocols in microbial ecology, highlighting key experimental considerations. Secondly, we review the applications of (RT)-Q-PCR analyses in environmental microbiology and evaluate the contribution and advances gained from such approaches. Finally, we conclude by offering future perspectives on the application of (RT)-Q-PCR in furthering understanding in microbial ecology, in particular, when coupled with other molecular approaches and more traditional investigations of environmental systems.

  11. Bioinformatic tools for PCR Primer design

    African Journals Online (AJOL)

    ES

    reaction (PCR), oligo hybridization and DNA sequencing. Proper primer design is actually one of the most important factors/steps in successful DNA sequencing. Various bioinformatics programs are available for selection of primer pairs from a template sequence. The plethora programs for PCR primer design reflects the.

  12. [E-MTAB-587] PCR_artifacts

    NARCIS (Netherlands)

    Muino Acuna, J.M.

    2011-01-01

    WARNING: This library was yield low amount of material and it was over-amplified by PCR. This libraries are used study the robustness of several statitical methods against PCR artifacts. ChIP experiments were performed on Arabidopsis wildtype inflorescences using an antibody raised against a

  13. Digital PCR for detection of citrus pathogens

    Science.gov (United States)

    Citrus trees are often infected with multiple pathogens of economic importance, especially those with insect or mite vectors. Real-time/quantitative PCR (qPCR) has been used for high-throughput detection and relative quantification of pathogens; however, target reference or standards are required. I...

  14. Testing for Genetically Modified Foods Using PCR

    Science.gov (United States)

    Taylor, Ann; Sajan, Samin

    2005-01-01

    The polymerase chain reaction (PCR) is a Nobel Prize-winning technique that amplifies a specific segment of DNA and is commonly used to test for the presence of genetic modifications. Students use PCR to test corn meal and corn-muffin mixes for the presence of a promoter commonly used in genetically modified foods, the cauliflower mosaic virus 35S…

  15. Validation of RNAi by real time PCR

    DEFF Research Database (Denmark)

    Josefsen, Knud; Lee, Ying Chiu

    2011-01-01

    Real time PCR is the analytic tool of choice for quantification of gene expression, while RNAi is concerned with downregulation of gene expression. Together, they constitute a powerful approach in any loss of function studies of selective genes. We illustrate here the use of real time PCR to verify...

  16. PCR specific for Actinobacillus pleuropneumoniae serotype 3

    DEFF Research Database (Denmark)

    Zhou, L.; Jones, S.C.P.; Angen, Øystein

    2008-01-01

    , but the method has liminations, for example, cross-reactions between serotypes 3, 6, and 8. This study describes the development of a serotype 3-specific PCR, based on the capsule locus, which can be used in a multiplex format with the organism's specific gene apxIV. The PCR test was evaluated on 266 strains...

  17. Aprendizaje temprano de arquitectura sustentable mediante vistas interiores graduadas

    Directory of Open Access Journals (Sweden)

    Adriana Edith Granero

    2014-06-01

    Full Text Available Lograr una arquitectura sustentable requiere, entre otros aspectos, incorporar consideraciones energéticas en etapas tempranas de los diseños arquitectónicos. Las definiciones ambientales involucran principalmente análisis numéricos y temporales, con un detalle de variables normalmente definidas en etapas avanzadas del trabajo proyectual, distantes de las resoluciones espaciales de la gestación inicial del diseño, especialmente durante el comienzo de la formación profesional. Se puede aplicar estos conocimientos técnicos en los alumnos, en etapas tempranas del proyecto, según las teorías de aprendizaje significativo y percepción, que sugieren capacidades de vinculación cognitiva, mediante tecnologías de información y simulaciones gráficas. Este artículo propone una secuencia de experiencias pedagógicas de integración, a partir de modelos para la resolución conjunta del diseño arquitectónico, que se enfoca en la elaboración de vistas espaciales graduadas según valores energéticos. Ello se ejemplifica con el dimensionamiento de vanos para iluminación natural en un recinto doméstico de trabajo, lo cual se relaciona con la privacidad y la visión exterior, como también con la comodidad y el consumo energético. El proceso se ensaya con grupos de estudiantes de arquitectura y a través de consultas con docentes y especialistas del área que reconocen una motivación significativa y de relación ambiental. Esta experiencia de integración gráfica implica una vinculación temprana de condiciones energéticas y espaciales, utilizable para la enseñanza y posterior desempeño profesional, que busca una arquitectura expresiva y ambientalmente responsable.

  18. Desarrollo de la lectura mediante estratégias integradoras

    Directory of Open Access Journals (Sweden)

    Solé, Maira

    2005-06-01

    Full Text Available La lectura y la escritura son procesos que cada día ameritan nuevos cambios y transformaciones. La propuesta de un Proyecto Pedagógico Integrador, (Fraca 2003 desarrollado con éxito en algunas instituciones venezolanas, se perfila como una alternativa significativa para el desarrollo de estos elementos. La idea o núcleo central es la integración de las diferentes asignaturas curriculares y lograr una globalización partiendo de sus objetivos y contenidos programáticos. El eje pedagógico integrador le permite al docente, evidenciar con mayor prontitud los resultados mediante actividades prácticas de lectura y escritura. Así mismo combina elementos claves del aprendizaje ausbeliano: información previa, información nueva y construcción de la información definitiva o integrada. La puesta en ejecución de las estrategias integradoras, en esta ocasión por maestros en formación (UNEG, a diferentes niños de escuelas del Estado Bolívar (Venezuela, certificando cómo la lectura y la escritura pueden tener un espacio ideal y significativo en la instrucción actual. Solo se necesita la intención, creatividad, dinamismo e ingenio. The reading and the writing plows processes that every day they require new changes and transformations. The proposal of an Integrative Pedagogic Project, (Fraca 2003 developed with success in some Venezuelan institutions; it is profiled like a significant alternative for the development of these elements. The idea or central nucleus is the integration of the different curricular subjects and to achieve a globalization leaving of its objectives and programmatic contents. The integrative pedagogic axis allows to the educational one, to evidence with more readiness the results by means of practical activities of reading and it notarizes. Likewise it combines key elements of the learning ausbeliano: previous information, new information and construction of the definitive or integrated information. The operation of

  19. Conocimientos, actitudes y prácticas sobre virus de papiloma humano (VPH y cáncer de cuello uterino en mujeres de 30 y más años de edad, de un barrio ribereño de Asunción, (Bañado Sur. 2012

    Directory of Open Access Journals (Sweden)

    Malvina Páez B

    2016-04-01

    Full Text Available Introducción: El cáncer de cuello uterino es un problema de salud pública en Paraguay. Objetivo: Determinar conocimientos, actitudes y prácticas sobre virus del papiloma humano (VPH y cáncer de cuello uterino en mujeres de 12 Unidades de Salud Familiar (USF de Bañado Sur-Asunción, periodo abril-octubre 2012. Metodología: Estudio descriptivo de corte transversal, utilizando cuestionario estructurado autoadministrado. Resultados: La edad promedio de las encuestadas fue 42 años, la mayoría en unión libre o casadas (70%; 65% tienen educación básica y media, 56% son amas de casa. El 83% tienen seguro médico; 78% escuchó hablar sobre cáncer de cuello uterino, 74% de éstas en los centros de salud. El 10% de las encuestadas conoce el VPH y lo relaciona con la enfermedad, 90 % escuchó hablar sobre la prueba de Papanicolaou, el 27 % de ellas sabe en qué consiste; 90% de las mujeres demostró actitud favorable y 56% prácticas favorables respecto a la prevención de la enfermedad. Conclusiones: El estudio permite conocer la percepción que tiene una población de mujeres de un barrio marginal de la capital del país, respecto al cáncer de cuello uterino y el principal factor de riesgo que lo produce, a fin de incrementar la prestación de servicios de prevención de este tipo de cáncer, además de propiciar el trabajo interinstitucional e intersectorial en la prevención y control de la enfermedad en el país.

  20. Polymerase chain reaction methods (PCR in agrobiotechnology

    Directory of Open Access Journals (Sweden)

    Taški-Ajduković Ksenija

    2006-01-01

    Full Text Available The agricultural biotechnology applies polymerase chain reaction (PCR technology at numerous steps throughout product development. The major uses of PCR technology during product development include gene discovery and cloning, vector construction, transformant identification, screening and characterization as well as seed quality control. Commodity and food companies as well as testing laboratories rely on PCR technology to verify the presence or absence of genetically modification (GM in a product or to quantify the amount of GM material present in the product. This article describes the fundamental elements of PCR analysis and its application to the testing of grains and highlights some of areas to which attention must be paid in order to produce reliable test results. The article also discuses issues related to the analysis of different matrixes and the effect they may have on the accuracy of the PCR analytical results.

  1. Mathematical analysis of the real time array PCR (RTA PCR) process

    NARCIS (Netherlands)

    Dijksman, Johan Frederik; Pierik, A.

    2012-01-01

    Real time array PCR (RTA PCR) is a recently developed biochemical technique that measures amplification curves (like with quantitative real time Polymerase Chain Reaction (qRT PCR)) of a multitude of different templates in a sample. It combines two different methods in order to profit from the

  2. Real-time PCR in virology.

    Science.gov (United States)

    Mackay, Ian M; Arden, Katherine E; Nitsche, Andreas

    2002-03-15

    The use of the polymerase chain reaction (PCR) in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting nucleic acids from a number of origins and it has become an essential tool in the research laboratory. Real-time PCR has engendered wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility and the reduced risk of carry-over contamination. There are currently five main chemistries used for the detection of PCR product during real-time PCR. These are the DNA binding fluorophores, the 5' endonuclease, adjacent linear and hairpin oligoprobes and the self-fluorescing amplicons, which are described in detail. We also discuss factors that have restricted the development of multiplex real-time PCR as well as the role of real-time PCR in quantitating nucleic acids. Both amplification hardware and the fluorogenic detection chemistries have evolved rapidly as the understanding of real-time PCR has developed and this review aims to update the scientist on the current state of the art. We describe the background, advantages and limitations of real-time PCR and we review the literature as it applies to virus detection in the routine and research laboratory in order to focus on one of the many areas in which the application of real-time PCR has provided significant methodological benefits and improved patient outcomes. However, the technology discussed has been applied to other areas of microbiology as well as studies of gene expression and genetic disease.

  3. Assessment of the real-time PCR and different digital PCR platforms for DNA quantification.

    Science.gov (United States)

    Pavšič, Jernej; Žel, Jana; Milavec, Mojca

    2016-01-01

    Digital PCR (dPCR) is beginning to supersede real-time PCR (qPCR) for quantification of nucleic acids in many different applications. Several analytical properties of the two most commonly used dPCR platforms, namely the QX100 system (Bio-Rad) and the 12.765 array of the Biomark system (Fluidigm), have already been evaluated and compared with those of qPCR. However, to the best of our knowledge, direct comparison between the three of these platforms using the same DNA material has not been done, and the 37 K array on the Biomark system has also not been evaluated in terms of linearity, analytical sensitivity and limit of quantification. Here, a first assessment of qPCR, the QX100 system and both arrays of the Biomark system was performed with plasmid and genomic DNA from human cytomegalovirus. With use of PCR components that alter the efficiency of qPCR, each dPCR platform demonstrated consistent copy-number estimations, which indicates the high resilience of dPCR. Two approaches, one considering the total reaction volume and the other considering the effective reaction size, were used to assess linearity, analytical sensitivity and variability. When the total reaction volume was considered, the best performance was observed with qPCR, followed by the QX100 system and the Biomark system. In contrast, when the effective reaction size was considered, all three platforms showed almost equal limits of detection and variability. Although dPCR might not always be more appropriate than qPCR for quantification of low copy numbers, dPCR is a suitable method for robust and reproducible quantification of viral DNA, and a promising technology for the higher-order reference measurement method.

  4. Material Biocompatibility for PCR Microfluidic Chips

    KAUST Repository

    Kodzius, Rimantas; Chang, Donald Choy; Gong, Xiuqing; Wen, Weijia; Wu, Jinbo; Xiao, Kang; Yi, Xin

    2010-01-01

    As part of the current miniaturization trend, biological reactions and processes are being adapted to microfluidics devices. PCR is the primary method employed in DNA amplification, its miniaturization is central to efforts to develop portable devices for diagnostics and testing purposes. A problem is the PCR-inhibitory effect due to interaction between PCR reagents and the surrounding environment, which effect is increased in high-surface-are-to-volume ration microfluidics. In this study, we evaluated the biocompatibility of various common materials employed in the fabrication of microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most of the cases, addition of bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, whereas they did show noticeable interaction with the DNA polymerase. Our test, instead of using microfluidic devices, can be easily conducted in common PCR tubes using a standard bench thermocycler. Our data supports an overview of the means by which the materials most bio-friendly to microfluidics can be selected.

  5. Material Biocompatibility for PCR Microfluidic Chips

    KAUST Repository

    Kodzius, Rimantas

    2010-04-23

    As part of the current miniaturization trend, biological reactions and processes are being adapted to microfluidics devices. PCR is the primary method employed in DNA amplification, its miniaturization is central to efforts to develop portable devices for diagnostics and testing purposes. A problem is the PCR-inhibitory effect due to interaction between PCR reagents and the surrounding environment, which effect is increased in high-surface-are-to-volume ration microfluidics. In this study, we evaluated the biocompatibility of various common materials employed in the fabrication of microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most of the cases, addition of bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, whereas they did show noticeable interaction with the DNA polymerase. Our test, instead of using microfluidic devices, can be easily conducted in common PCR tubes using a standard bench thermocycler. Our data supports an overview of the means by which the materials most bio-friendly to microfluidics can be selected.

  6. Comparison between digital PCR and real-time PCR in detection of Salmonella typhimurium in milk.

    Science.gov (United States)

    Wang, Meng; Yang, Junjie; Gai, Zhongtao; Huo, Shengnan; Zhu, Jianhua; Li, Jun; Wang, Ranran; Xing, Sheng; Shi, Guosheng; Shi, Feng; Zhang, Lei

    2018-02-02

    As a kind of zero-tolerance foodborne pathogens, Salmonella typhimurium poses a great threat to quality of food products and public health. Hence, rapid and efficient approaches to identify Salmonella typhimurium are urgently needed. Combined with PCR and fluorescence technique, real-time PCR (qPCR) and digital PCR (ddPCR) are regarded as suitable tools for detecting foodborne pathogens. To compare the effect between qPCR and ddPCR in detecting Salmonella typhimurium, a series of nucleic acid, pure strain culture and spiking milk samples were applied and the resistance to inhibitors referred in this article as well. Compared with qPCR, ddPCR exhibited more sensitive (10 -4 ng/μl or 10 2 cfu/ml) and less pre-culturing time (saving 2h). Moreover, ddPCR had stronger resistance to inhibitors than qPCR, yet absolute quantification hardly performed when target's concentration over 1ng/μl or 10 6 cfu/ml. This study provides an alternative strategy in detecting foodborne Salmonella typhimurium. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Real-time PCR in virology

    OpenAIRE

    Mackay, Ian M.; Arden, Katherine E.; Nitsche, Andreas

    2002-01-01

    The use of the polymerase chain reaction (PCR) in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting nucleic acids from a number of origins and it has become an essential tool in the research laboratory. Real-time PCR has engendered wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility and the reduced risk of carry-over contamination. There are currently five main chemistries used for the detection of P...

  8. Calibrated user-friendly reverse transcriptase-PCR assay

    DEFF Research Database (Denmark)

    Bor, M V; Sørensen, B S; Rammer, P

    1998-01-01

    We report a competitive reverse transcriptase-PCR (RT-PCR) assay and a calibrated user-friendly RT-PCR assay (CURT-PCR) for epidermal growth factor receptor (EGFR) mRNA. A calibrator was prepared from isolated rat liver RNA, and the amount of EGFR mRNA was determined by competitive RT-PCR. In CUR...

  9. Transgene detection by digital droplet PCR.

    Directory of Open Access Journals (Sweden)

    Dirk A Moser

    Full Text Available Somatic gene therapy is a promising tool for the treatment of severe diseases. Because of its abuse potential for performance enhancement in sports, the World Anti-Doping Agency (WADA included the term 'gene doping' in the official list of banned substances and methods in 2004. Several nested PCR or qPCR-based strategies have been proposed that aim at detecting long-term presence of transgene in blood, but these strategies are hampered by technical limitations. We developed a digital droplet PCR (ddPCR protocol for Insulin-Like Growth Factor 1 (IGF1 detection and demonstrated its applicability monitoring 6 mice injected into skeletal muscle with AAV9-IGF1 elements and 2 controls over a 33-day period. A duplex ddPCR protocol for simultaneous detection of Insulin-Like Growth Factor 1 (IGF1 and Erythropoietin (EPO transgenic elements was created. A new DNA extraction procedure with target-orientated usage of restriction enzymes including on-column DNA-digestion was established. In vivo data revealed that IGF1 transgenic elements could be reliably detected for a 33-day period in DNA extracted from whole blood. In vitro data indicated feasibility of IGF1 and EPO detection by duplex ddPCR with high reliability and sensitivity. On-column DNA-digestion allowed for significantly improved target detection in downstream PCR-based approaches. As ddPCR provides absolute quantification, it ensures excellent day-to-day reproducibility. Therefore, we expect this technique to be used in diagnosing and monitoring of viral and bacterial infection, in detecting mutated DNA sequences as well as profiling for the presence of foreign genetic material in elite athletes in the future.

  10. Diagnosis of trichomonas vaginalis infection by PCR

    International Nuclear Information System (INIS)

    Issa, R.M.; Shalaby, M.A.

    2007-01-01

    To compare the sensitivity of PCR, wet preparation and culture in detecting Trichomonas vaginalis in urine and vaginal fluid. A PCR targeting the beta-tubulin genes of T. vaginalis was used for the detection of the organism in both vaginal swab and urine specimens from infected patients. Random urine samples were collected from 30 patients (23 females and 7 males), and tested for T. vaginalis by wet preparation and the Inpouch T. vaginalis culture systeme. Two vaginal swabs were collected by each woman. PCR detection. was carried out on samples negative by first methods. The positive result was found in 28.57% in male urine and 39.13% in female urine samples, 65.21% in 1st swab and 78.26 % in 2nd swab by wet preparation. By culture, the male urine samples showed 42.85% positive, female urine 69.56% while 1st swab showed 86.95% positive and 2nd swab 91.30% positive. All negative cases by culture in urine and vaginal samples were tested by PCR, which showed 2 cases to be positive in male urine samples and 5 cases positive in female urine sample. PCR assay was as good as or more sensitive than wet preparation and culture and resulted in practical advantage of providing results in shorter time. However, PCR test is still very expensive. (author)

  11. Ensayo no destructivo de soldaduras en pernos conectores mediante inspección acústica

    OpenAIRE

    Aznar, A.; Cervera, J.; Ortiz, J.; Hernando, J. I.

    2012-01-01

    Los pernos conectores aportan múltiples ventajas de uso, entre las que se encuentra el elevado margen de seguridad que ofrecen sus soldaduras ejecutadas mediante arco eléctrico. Estas soldaduras, aunque ampliamente fiables, son difícilmente comprobadas mediante ensayos no destructivos. Aparte de la inspección visual, que aporta gran información sobre la calidad de ejecución de la soldadura, el resto de ensayos no destructivos (líquidos penetrantes, partículas magnéticas, ultrasonidos, radiogr...

  12. Modulación del crecimiento vertebral mediante electrocoagulación hemicircunferencial vertebral asistida

    OpenAIRE

    Caballero García, Alberto

    2011-01-01

    Nuestro trabajo está basado en la posibilidad de controlar el desarrollo asimétrico de los cartílagos de crecimiento vertebral, mediante la realización de una fisiodesis hemivertebral, con electrocoagulación, videoasistida por toracoscópica. Se realizará en cinco niveles torácicos, con un abordaje anterior mínimamente invasivo. Por lo tanto, planteamos como hipótesis de trabajo que La destrucción de las fisis de crecimiento vertebral mediante electrocoagulación, videoasistida por vía toracosc...

  13. Modelado de un amortiguador magneto-reológico mediante EcosimPro

    OpenAIRE

    Rodríguez Cadenas, Rubén

    2012-01-01

    El objetivo de este proyecto es la creación de una librería en la herramienta de modelado y simulación EcosimPro enfocada a amortiguadores magneto‐reológicos. El modelado y simulación mediante cualquier herramienta informática permite la obtención de datos y el desarrollo de componentes con un coste inferior al que habría que invertir mediante una experimentación real. Además, permite llevar el componente hasta el límite sin el riesgo de romperlo o dejarlo inutilizable. Por tanto, se puede de...

  14. SASqPCR: robust and rapid analysis of RT-qPCR data in SAS.

    Directory of Open Access Journals (Sweden)

    Daijun Ling

    Full Text Available Reverse transcription quantitative real-time PCR (RT-qPCR is a key method for measurement of relative gene expression. Analysis of RT-qPCR data requires many iterative computations for data normalization and analytical optimization. Currently no computer program for RT-qPCR data analysis is suitable for analytical optimization and user-controllable customization based on data quality, experimental design as well as specific research aims. Here I introduce an all-in-one computer program, SASqPCR, for robust and rapid analysis of RT-qPCR data in SAS. This program has multiple macros for assessment of PCR efficiencies, validation of reference genes, optimization of data normalizers, normalization of confounding variations across samples, and statistical comparison of target gene expression in parallel samples. Users can simply change the macro variables to test various analytical strategies, optimize results and customize the analytical processes. In addition, it is highly automatic and functionally extendable. Thus users are the actual decision-makers controlling RT-qPCR data analyses. SASqPCR and its tutorial are freely available at http://code.google.com/p/sasqpcr/downloads/list.

  15. PCR+ In Diesel Fuels and Emissions Research

    Energy Technology Data Exchange (ETDEWEB)

    McAdams, H.T.

    2002-04-15

    In past work for the U.S. Department of Energy (DOE) and Oak Ridge National Laboratory (ORNL), PCR+ was developed as an alternative methodology for building statistical models. PCR+ is an extension of Principal Components Regression (PCR), in which the eigenvectors resulting from Principal Components Analysis (PCA) are used as predictor variables in regression analysis. The work was motivated by the observation that most heavy-duty diesel (HDD) engine research was conducted with test fuels that had been ''concocted'' in the laboratory to vary selected fuel properties in isolation from each other. This approach departs markedly from the real world, where the reformulation of diesel fuels for almost any purpose leads to changes in a number of interrelated properties. In this work, we present new information regarding the problems encountered in the conventional approach to model-building and how the PCR+ method can be used to improve research on the relationship between fuel characteristics and engine emissions. We also discuss how PCR+ can be applied to a variety of other research problems related to diesel fuels.

  16. Método de eliminación de trihalometanos y/o contaminantes emergentes mediante plasma

    OpenAIRE

    Erra Serrabasa, Pilar; Jover Comas, Eric; Molina Mansilla, Ricardo; Bertrán Serra, Enric; Bayona Termens, Josep María; Reyes Contreras, Carolina

    2009-01-01

    Método de eliminación de trihalometanos y/o contaminantes emergentes mediante plasma. Se describe un método de eliminación de trihalometanos y contaminantes refractarios en medios acuosos mediante la aplicación directa de plasma para conseguir la degradación de los compuestos contaminantes presentes en el agua.

  17. Detección y tipificación de virus del papiloma humano en biopsias de carcinoma ductal infiltrante y lesiones benignas de mama en mujeres venezolanas

    OpenAIRE

    Solorzano, Marisé; Bastidas, Marco; Quintero, Militza; Rojas, Lisbeth; Stea, Domingo; Villasmil, Saúl; Acosta, Víctor; Marín, Carmen; Ramírez, Ana; Blanco, Natasha; Cruz, Jhon; Puig, Juan

    2016-01-01

    Objetivo: Realizar la detección y tipificación del virus del papiloma humano (VPH) en muestras de biopsias de tejido mamario con carcinoma ductal infiltrante. Métodos: Estudio descriptivo de corte transversal de 57 biopsias de carcinoma ductal infiltrante, y 41 biopsias de lesiones benignas de mama de pacientes venezolanas, estas fueron evaluadas utilizando la técnica PCR-RFLP en busca de la presencia del genoma del virus de papiloma humano. El riesgo OR fue evaluado mediante análisis estadís...

  18. Establecer las condiciones necesarias para procesar materiales termoestables mediante el rotomoldeo

    OpenAIRE

    Pérez O., Daniel

    2009-01-01

    En este trabajo se establecieron las condiciones necesarias para procesar materiales termoestables mediante la técnica de rotomoldeo, comenzando por el estudio de las condiciones de curado y viscosidad relativa, donde se evidenció una relación directa del porcentaje de catalizador en función del tiempo y la temperatura de polimerización.

  19. Variabilidad genética de poblaciones en cautiverio de Crocodylus moreletii (Crocodylia: Crocodylidae mediante el uso de marcadores microsatelitales

    Directory of Open Access Journals (Sweden)

    Ricardo Serna-Lagunes

    2012-03-01

    Full Text Available Crocodylus moreletii representa un emblema para los ecosistemas tropicales de México pero actualmente está amenazada por extinción. Sorprendentemente, hay una falta de información de su constitución genética, que debe ser evaluada para un manejo apropiado ex situ y para toma de decisiones en la liberación de cocodrilos a su hábitat natural. El objetivo del estudio fue caracterizar y comparar la variabilidad genética de cuatro grupos poblacionales de C. moreletii (dos silvestres y dos nacidas ex situ. Mediante PCR se amplificaron siete loci de microsatélites polimórficos, sin embargo se encontró déficit de heterocigotos en las poblaciones (promedio H O=0.02 mermado por la presencia de alelos nulos. El AMOVA indicó que la mayor proporción de variabilidad genética se encuentra dentro de las poblaciones y una limitada diferenciación genética entre poblaciones (promedio F ST =0.03, probablemente debida al alto índice de endogamia (promedio F IS=0.97. Al comparar la variabilidad genética inter e intra especies de cocodrilianos, encontramos que en C. moreletii está muy por debajo de los reportados. Se concluye que la limitada variabilidad genética de las poblaciones nacidas ex situ probablemente se debe al efecto fundador derivado de la estructura social de sus progenitores, y de las poblaciones silvestres, por el efecto cuello de botella, inferido por el limitado tamaño efectivo de población que presentó históricamente en su distribución natural.

  20. From the 'PCR' function to the 'PCR' profession; de la fonction 'PCR' au metier 'PCR'

    Energy Technology Data Exchange (ETDEWEB)

    Perrin, L. [CERAP, 91 - Gif sur Yvette (France)

    2008-07-01

    After having recalled the legal context concerning the appointment and training of a radiation protection expert (PCR for 'personne competente en radioprotection'), the author outlines that the PCR's role has notably evolved: his function is now of primary importance in the company and his activity does not correspond to the legal framework any longer. Moreover, with the application of a European directive, some small establishments possessing ionizing radiation sources are disadvantaged, and the PCR is now facing an increasing number of missions and tasks. The author gives a list of them and assesses a needed time of 146 days per year: this means PCRs cannot have an other activity within their company

  1. PCR melting profile (PCR MP - a new tool for differentiation of Candida albicans strains

    Directory of Open Access Journals (Sweden)

    Nowak Magdalena

    2009-11-01

    Full Text Available Abstract Background We have previously reported the use of PCR Melting Profile (PCR MP technique based on using low denaturation temperatures during ligation mediated PCR (LM PCR for bacterial strain differentiation. The aim of the current study was to evaluate this method for intra-species differentiation of Candida albicans strains. Methods In total 123 Candida albicans strains (including 7 reference, 11 clinical unrelated, and 105 isolates from patients of two hospitals in Poland were examined using three genotyping methods: PCR MP, macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis (REA-PFGE and RAPD techniques. Results The genotyping results of the PCR MP were compared with results from REA-PFGE and RAPD techniques giving 27, 26 and 25 unique types, respectively. The results showed that the PCR MP technique has at least the same discriminatory power as REA-PFGE and RAPD. Conclusion Data presented here show for the first time the evaluation of PCR MP technique for candidial strains differentiation and we propose that this can be used as a relatively simple and cheap technique for epidemiological studies in short period of time in hospital.

  2. Propidium monoazide reverse transcription PCR and RT-qPCR for detecting infectious enterovirus and norovirus

    Science.gov (United States)

    Presently there is no established cell line or small animal model that allows for the detection of infectious human norovirus. Current methods based on RT-PCR and RT-qPCR detect both infectious and non-infectious virus and thus the conclusions that may be drawn regarding the publ...

  3. Detection of Leishmania infantum in animals and their ectoparasites by conventional PCR and real time PCR.

    Science.gov (United States)

    de Morais, Rayana Carla Silva; Gonçalves, Suênia da Cunha; Costa, Pietra Lemos; da Silva, Kamila Gaudêncio; da Silva, Fernando José; Silva, Rômulo Pessoa E; de Brito, Maria Edileuza Felinto; Brandão-Filho, Sinval Pinto; Dantas-Torres, Filipe; de Paiva-Cavalcanti, Milena

    2013-04-01

    Visceral leishmaniosis (VL) is a parasitic disease caused by Leishmania infantum, which is primarily transmitted by phlebotomine sandflies. However, there has been much speculation on the role of other arthropods in the transmission of VL. Thus, the aim of this study was to assess the presence of L. infantum in cats, dogs and their ectoparasites in a VL-endemic area in northeastern Brazil. DNA was extracted from blood samples and ectoparasites, tested by conventional PCR (cPCR) and quantitative real time PCR (qPCR) targeting the L. infantum kinetoplast DNA. A total of 280 blood samples (from five cats and 275 dogs) and 117 ectoparasites from dogs were collected. Animals were apparently healthy and not previously tested by serological or molecular diagnostic methods. Overall, 213 (76.1 %) animals and 51 (43.6 %) ectoparasites were positive to L. infantum, with mean parasite loads of 795.2, 31.9 and 9.1 fg in dogs, cats and ectoparasites, respectively. Concerning the positivity between dogs and their ectoparasites, 32 (15.3 %) positive dogs were parasitized by positive ectoparasites. The overall concordance between the PCR protocols used was 59.2 %, with qPCR being more efficient than cPCR; 34.1 % of all positive samples were exclusively positive by qPCR. The high number of positive animals and ectoparasites also indicates that they could serve as sentinels or indicators of the circulation of L. infantum in risk areas.

  4. Diagnosis of Cutaneous Leishmaniasis by Multiplex PCR

    Directory of Open Access Journals (Sweden)

    M Heiat

    2010-07-01

    Full Text Available Introduction: Annually, more than 14 million people are reported to be infected with Leishmaniasis all over the world. In Iran, this disease is seen in the form of cutaneous and visceral leishmaniasis, of which the cutaneous form is more wide spread. In recent years, cutaneous leishmaniaisis is diagnosed by PCR utilizing specific primers in order to amplify different parasite genes including ribosomal RNA genes, kinetoplast DNA or tandem repeating sequences. The aim of this research was to detect early stage cutaneous leishmaniasis using Multiplex-PCR technique. Methods: In this study, 67 samples were prepared from patients with cutaneous leishmaniasis. DNA was extracted with phenolchloroform. Each specimen was analyzed using two different pairs of PCR primers. The sensitivity of each PCR was optimized on pure Leishmania DNA prior to use for diagnosis. Two standard parasites L. major and L. tropica were used as positive control. Results: DNA amplification fragments were two 115 bp and 683 bp for AB and UL primers, respectively. The sensitivity of two primers was not equal for detection of L. major and L. tropica. The sensivity of PCR with AB primer was 35 cells, while that for UL primer was 40 cells. Conclusion: The results of this study indicate that PCR is a sensitive diagnostic assay for cutaneous leishmaniasis and could be employed as the new standard for routine diagnosis when species identification is not required. However, the ability to identify species is especially important in prognosis of the disease and in deciding appropriate therapy, especially in regions where more than one type of species and disease are seen by clinicians.

  5. Detección y cuantificación del Potato mop-top virus (PMTV) en Colombia mediante qRT-PCR

    OpenAIRE

    Nevar García Bastidas; Pablo Gutiérrez Sánchez; Mauricio Marín Montoya

    2013-01-01

    El Potato mop-top virus (PMTV) es uno de los virus re-emergentes en cultivos de papa en Colombia. Es transmitido por Spongospora subterranea, el agente causal de la sarna polvosa. La detección del PMTV presenta dificultades debido a su distribución irregular en las plantas, bajo título y movimiento sistémico como ARN desnudo. Con el fin de ampliar el rango de herramientas disponibles para detectar el PMTV en los programas de certificación de tubérculo-semilla, en este estudio se evaluó la pru...

  6. Reduction of heteroduplex formation in PCR amplification

    Czech Academy of Sciences Publication Activity Database

    Michu, Elleni; Mráčková, Martina; Vyskot, Boris; Žlůvová, Jitka

    2010-01-01

    Roč. 54, č. 1 (2010), s. 173-176 ISSN 0006-3134 R&D Projects: GA AV ČR(CZ) KJB600040801; GA ČR(CZ) GD204/09/H002; GA AV ČR(CZ) IAA600040801; GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : polymerase chain reaction * reconditioning PCR * mixed-template PCR Subject RIV: BO - Biophysics Impact factor: 1.582, year: 2010

  7. A naked-eye colorimetric "PCR developer"

    Science.gov (United States)

    Valentini, Paola; Pompa, Pier Paolo

    2016-04-01

    Despite several advances in molecular biology and diagnostics, Polymerase Chain Reaction (PCR) is currently the gold standard for nucleic acids amplification and detection, due to its versatility, low-cost and universality, with estimated genetically modified organisms, and pathogens). The PCR developer proved to be highly specific and ultra-sensitive, discriminating down to few copies of HIV viral DNA, diluted in an excess of interfering human genomic DNA, which is a clinically relevant viral load. Hence, it could be a valuable tool for both academic research and clinical applications.

  8. Detección de Treponema pallidum subespecie pallidum para el diagnóstico de sífilis congénita mediante reacción en cadena de la polimerasa anidada.

    Science.gov (United States)

    Pinilla, Gladys; Campos, Lesly; Durán, Andrea; Navarrete, Jeannette; Muñoz, Liliana

    2018-03-15

    Introducción. La sífilis es una enfermedad producida por Treponema pallidum subespecie pallidum cuya incidencia mundial es de 12 millones de casos por año, aproximadamente; de estos, más de dos millones se presentan en mujeres gestantes, siendo la sífilis congénita la complicación más grave de esta infección en el embarazo.Objetivo. Detectar la presencia de T. pallidum subespecie pallidum en muestras clínicas para el diagnóstico de sífilis congénita mediante reacción en cadena de la polimerasa (PCR) anidada y determinar su concordancia con las pruebas serológicas.Materiales y métodos. Mediante PCR convencional y anidada, se amplificaron tres genes diana (polA, 16S ADNr y TpN47) y se confirmaron los productos de amplificación de los genes TpN47 y polA por secuenciación. Las pruebas serológicas empleadas fueron la VDRL (Venereal Disease Research Laboratory), la de reagina plasmática rápida (Rapid Plasma Reagin, RPR) y la de aglutinación de partículas para Treponema pallidum (Treponema pallidum Particle Agglutination Assay, TPPA).Resultados. La sensibilidad para la PCR convencional fue de 52 pg y, para la PCR anidada, de 0,52 pg. La especificidad con los iniciadores TpN47 y polA fue de 100 %; los resultados de la secuenciación mostraron una identidad de 97 % con T. pallidum. En 70 % de las muestras, los resultados de las pruebas serológicas y la PCR anidada concordaron.Conclusión. El gen TpN47 resultó ser el mejor blanco molecular para la identificación de T. pallidum. La PCR anidada se presenta como una alternativa de diagnóstico molecular promisoria para el diagnóstico de sífilis congénita.

  9. Detection of Mycobacterium Tuberculosis by using PCR

    International Nuclear Information System (INIS)

    Suhadi, F; Dadang-Sudrajat; Maria-Lina, R.

    1996-01-01

    Polymerase Chain Reaction (PCR) procedure using three primary set derived from repetitive DNA sequence specific to mycobacteria was used to diagnose pathogenic Mycobacterium tuberculosis. The assay was specific for M. tuberculosis and could be used to detect the amount DNA less than 10 -9 g

  10. PCR detection of potato cyst nematode.

    Science.gov (United States)

    Reid, Alex

    2009-01-01

    Potato cyst nematode (PCN) is responsible for losses in potato production totalling millions of euros every year in the EC. It is important for growers to know which species is present in their land as this determines its subsequent use. The two species Globodera pallida and Globodera rostochiensis can be differentiated using an allele-specific PCR.

  11. Real-time PCR gene expression profiling

    Czech Academy of Sciences Publication Activity Database

    Kubista, Mikael; Sjögreen, B.; Forootan, A.; Šindelka, Radek; Jonák, Jiří; Andrade, J.M.

    2007-01-01

    Roč. 1, - (2007), s. 56-60 ISSN 1360-8606 R&D Projects: GA AV ČR KJB500520601 Institutional research plan: CEZ:AV0Z50520514 Keywords : real - time PCR, * expression profiling * statistical analysis Subject RIV: EB - Genetics ; Molecular Biology

  12. A survey of tools for the analysis of quantitative PCR (qPCR) data.

    Science.gov (United States)

    Pabinger, Stephan; Rödiger, Stefan; Kriegner, Albert; Vierlinger, Klemens; Weinhäusel, Andreas

    2014-09-01

    Real-time quantitative polymerase-chain-reaction (qPCR) is a standard technique in most laboratories used for various applications in basic research. Analysis of qPCR data is a crucial part of the entire experiment, which has led to the development of a plethora of methods. The released tools either cover specific parts of the workflow or provide complete analysis solutions. Here, we surveyed 27 open-access software packages and tools for the analysis of qPCR data. The survey includes 8 Microsoft Windows, 5 web-based, 9 R-based and 5 tools from other platforms. Reviewed packages and tools support the analysis of different qPCR applications, such as RNA quantification, DNA methylation, genotyping, identification of copy number variations, and digital PCR. We report an overview of the functionality, features and specific requirements of the individual software tools, such as data exchange formats, availability of a graphical user interface, included procedures for graphical data presentation, and offered statistical methods. In addition, we provide an overview about quantification strategies, and report various applications of qPCR. Our comprehensive survey showed that most tools use their own file format and only a fraction of the currently existing tools support the standardized data exchange format RDML. To allow a more streamlined and comparable analysis of qPCR data, more vendors and tools need to adapt the standardized format to encourage the exchange of data between instrument software, analysis tools, and researchers.

  13. Quantitative Real-Time PCR using the Thermo Scientific Solaris qPCR Assay

    Science.gov (United States)

    Ogrean, Christy; Jackson, Ben; Covino, James

    2010-01-01

    The Solaris qPCR Gene Expression Assay is a novel type of primer/probe set, designed to simplify the qPCR process while maintaining the sensitivity and accuracy of the assay. These primer/probe sets are pre-designed to >98% of the human and mouse genomes and feature significant improvements from previously available technologies. These improvements were made possible by virtue of a novel design algorithm, developed by Thermo Scientific bioinformatics experts. Several convenient features have been incorporated into the Solaris qPCR Assay to streamline the process of performing quantitative real-time PCR. First, the protocol is similar to commonly employed alternatives, so the methods used during qPCR are likely to be familiar. Second, the master mix is blue, which makes setting the qPCR reactions easier to track. Third, the thermal cycling conditions are the same for all assays (genes), making it possible to run many samples at a time and reducing the potential for error. Finally, the probe and primer sequence information are provided, simplifying the publication process. Here, we demonstrate how to obtain the appropriate Solaris reagents using the GENEius product search feature found on the ordering web site (www.thermo.com/solaris) and how to use the Solaris reagents for performing qPCR using the standard curve method. PMID:20567213

  14. Pneumocystis PCR: It Is Time to Make PCR the Test of Choice.

    Science.gov (United States)

    Doyle, Laura; Vogel, Sherilynn; Procop, Gary W

    2017-01-01

    The testing strategy for Pneumocystis at the Cleveland Clinic changed from toluidine blue staining to polymerase chain reaction (PCR). We studied the differences in positivity rates for these assays and compared each with the detection of Pneumocystis in companion specimens by cytology and surgical pathology. We reviewed the results of all Pneumocystis test orders 1 year before and 1 year after the implementation of a Pneumocystis -specific PCR. We also reviewed the corresponding cytology and surgical pathology results, if performed. Finally, we reviewed the medical records of patients with rare Pneumocystis detected by PCR in an effort to differentiate colonization vs true disease. Toluidine blue staining and surgical pathology had similar sensitivities and negative predictive values, both of which were superior to cytology. There was a >4-fold increase in the annual detection of Pneumocystis by PCR compared with toluidine blue staining (toluidine blue staining: 11/1583 [0.69%] vs PCR: 44/1457 [3.0%]; chi-square P < .001). PCR detected 1 more case than surgical pathology and was far more sensitive than cytology. Chart review demonstrated that the vast majority of patients with rare Pneumocystis detected were immunosuppressed, had radiologic findings supportive of this infection, had no other pathogens detected, and were treated for pneumocystosis by the clinical team. PCR was the most sensitive method for the detection of Pneumocystis and should be considered the diagnostic test of choice. Correlation with clinical and radiologic findings affords discrimination of early true disease from the far rarer instances of colonization.

  15. Real-time PCR (qPCR) primer design using free online software.

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most commonly used fluorescent chemistries are SYBR® Green dyes and TaqMan®, Molecular Beacon or Scorpion probes. SYBR® Green is very simple to use and cost efficient. As SYBR® Green dye binds to any double-stranded DNA product, its success depends greatly on proper primer design. Many types of online primer design software are available, which can be used free of charge to design desirable SYBR® Green-based qPCR primers. This laboratory exercise is intended for those who have a fundamental background in PCR. It addresses the basic fluorescent chemistries of real-time PCR, the basic rules and pitfalls of primer design, and provides a step-by-step protocol for designing SYBR® Green-based primers with free, online software. Copyright © 2010 Wiley Periodicals, Inc.

  16. Digital PCR for direct quantification of viruses without DNA extraction

    OpenAIRE

    Pav?i?, Jernej; ?el, Jana; Milavec, Mojca

    2015-01-01

    DNA extraction before amplification is considered an essential step for quantification of viral DNA using real-time PCR (qPCR). However, this can directly affect the final measurements due to variable DNA yields and removal of inhibitors, which leads to increased inter-laboratory variability of qPCR measurements and reduced agreement on viral loads. Digital PCR (dPCR) might be an advantageous methodology for the measurement of virus concentrations, as it does not depend on any calibration mat...

  17. Comparison of simultaneous splenic sample PCR with blood sample PCR for diagnosis and treatment of experimental Ehrlichia canis infection.

    Science.gov (United States)

    Harrus, Shimon; Kenny, Martin; Miara, Limor; Aizenberg, Itzhak; Waner, Trevor; Shaw, Susan

    2004-11-01

    This report presents evidence that dogs recover from acute canine monocytic ehrlichiosis (CME) after 16 days of doxycycline treatment (10 mg/kg of body weight every 24 h). Blood PCR was as valuable as splenic aspirate PCR for early diagnosis of acute CME. Splenic aspirate PCR was, however, superior to blood PCR for the evaluation of ehrlichial elimination.

  18. Detection of hepatitis C virus RNA: comparison of one-stage polymerase chain reaction (PCR) with nested-set PCR.

    OpenAIRE

    Gretch, D R; Wilson, J J; Carithers, R L; dela Rosa, C; Han, J H; Corey, L

    1993-01-01

    We evaluated a new hepatitis C virus RNA assay based on one-stage PCR followed by liquid hybridization with an oligonucleotide probe and compared it with nested-set PCR. The one-stage and nested-set PCR assays had identical sensitivities in analytical experiments and showed 100% concordance when clinical specimens were used. One-stage PCR may be less prone to contamination than nested-set PCR.

  19. Programación por metas Energía alternativa mediante biomasa.

    Directory of Open Access Journals (Sweden)

    Guerrero Casas, Flor María

    2003-01-01

    Full Text Available En este trabajo se presenta un modelo multicriterio de localización de centrales de generación de energía eléctrica mediante biomasa. Los objetivos considerados son: (1 minimizar el coste total de la operación, (2 maximizar la producción de electricidad obtenida, (3 maximizar la distancia entre plantas, (4 maximizar la aceptación social y (5 establecer las plantas o ampliaciones en aquellos lugares donde exista una mayor predisposición por parte de las administraciones locales. Finalmente, se concluye con una aplicación práctica mediante programación por metas ponderadas para la región andaluza, considerando los residuos procedentes del olivar como fuente de energía.

  20. Application of droplet digital PCR for quantitative detection of Spiroplasma citri in comparison with real time PCR.

    Directory of Open Access Journals (Sweden)

    Yogita Maheshwari

    Full Text Available Droplet digital polymerase chain reaction (ddPCR is a method for performing digital PCR that is based on water-oil emulsion droplet technology. It is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. This study evaluated the applicability of ddPCR as a quantitative detection tool for the Spiroplasma citri, causal agent of citrus stubborn disease (CSD in citrus. Two sets of primers, SP1, based on the spiral in housekeeping gene, and a multicopy prophage gene, SpV1 ORF1, were used to evaluate ddPCR in comparison with real time (quantitative PCR (qPCR for S. citri detection in citrus tissues. Standard curve analyses on tenfold dilution series showed that both ddPCR and qPCR exhibited good linearity and efficiency. However, ddPCR had a tenfold greater sensitivity than qPCR and accurately quantified up to one copy of spiralin gene. Receiver operating characteristic analysis indicated that the ddPCR methodology was more robust for diagnosis of CSD and the area under the curve was significantly broader compared to qPCR. Field samples were used to validate ddPCR efficacy and demonstrated that it was equal or better than qPCR to detect S. citri infection in fruit columella due to a higher pathogen titer. The ddPCR assay detected both the S. citri spiralin and the SpV1 ORF1 targets quantitatively with high precision and accuracy compared to qPCR assay. The ddPCR was highly reproducible and repeatable for both the targets and showed higher resilience to PCR inhibitors in citrus tissue extract for the quantification of S. citri compare to qPCR.

  1. Clostridium perfringens isolate typing by multiplex PCR

    Directory of Open Access Journals (Sweden)

    MR Ahsani

    2010-01-01

    Full Text Available Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39% type A, 21.74% type B, 34.78% type C and 26.09% type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.

  2. Species identification and quantification in meat and meat products using droplet digital PCR (ddPCR).

    Science.gov (United States)

    Floren, C; Wiedemann, I; Brenig, B; Schütz, E; Beck, J

    2015-04-15

    Species fraud and product mislabelling in processed food, albeit not being a direct health issue, often results in consumer distrust. Therefore methods for quantification of undeclared species are needed. Targeting mitochondrial DNA, e.g. CYTB gene, for species quantification is unsuitable, due to a fivefold inter-tissue variation in mtDNA content per cell resulting in either an under- (-70%) or overestimation (+160%) of species DNA contents. Here, we describe a reliable two-step droplet digital PCR (ddPCR) assay targeting the nuclear F2 gene for precise quantification of cattle, horse, and pig in processed meat products. The ddPCR assay is advantageous over qPCR showing a limit of quantification (LOQ) and detection (LOD) in different meat products of 0.01% and 0.001%, respectively. The specificity was verified in 14 different species. Hence, determining F2 in food by ddPCR can be recommended for quality assurance and control in production systems. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  3. Conhecimento, atitude e práticas na prevenção do câncer de colo uterino e hpv em adolescentes Conocimientos, actitudes y prácticas en la prevención de cáncer cervicouterino y el VPH en adolescentes Knowledge, attitude and practice in the prevention of cervical cancer and HPV in adolescents

    Directory of Open Access Journals (Sweden)

    Ferla Maria Simas Bastos Cirino

    2010-03-01

    Full Text Available O início sexual cada vez mais cedo propicia alta vulnerabilidade da adolescente a problemas da esfera sexual/reprodutiva, incluindo o câncer de colo uterino e a infecção pelo HPV. O presente estudo teve como objetivo identificar o conhecimento, atitude e prática na prevenção do câncer de colo uterino e infecção pelo HPV na população adolescente e avaliar as situações que as tornam vulneráveis. Trata-se de estudo transversal realizado em uma escola pública de São Paulo com 134 adolescentes entre 14 e 19 anos. Verificou-se idade de iniciação sexual aos 14,8 anos em média. Grande parte das adolescentes não apresentou conhecimento adequado sobre a prevenção desta neoplasia. A adesão ao Papanicolaou também se mostrou baixa. As estatísticas justificam a inserção da adolescente nos programas de detecção deste câncer. É preciso haver investimentos na educação sexual nas instituições de ensino e associar campanhas de Papanicolaou com atividades educativas, com enfoque adequado e linguagem apropriada.La iniciación sexual cada vez más temprana propicia un nivel alto de vulnerabilidad de los adolescentes a los problemas de la esfera sexual / reproductiva, incluyendo el cáncer del cuello del útero y la infección por VPH. Este estudio tuvo como objetivo identificar los conocimientos, actitudes y prácticas en la prevención de cáncer cervical y la infección por VPH en la población adolescente y evaluar las situaciones que los tornan vulnerables. Este es un estudio transversal realizado en una escuela pública en Sao Paulo con 134 adolescentes entre 14 y 19 años. El promedio de la edad de iniciación sexual fue de 14,8 años. La mayoría de los adolescentes no tienen conocimientos adecuados sobre la prevención de este tipo de cáncer. También presentó bajas tasas de la prueba del Papanicolaou . Las estadísticas justifican la inclusión de estas adolescentes en los programas para la detección de este cáncer. Es

  4. Analysis of extracellular RNA by digital PCR

    Directory of Open Access Journals (Sweden)

    Kenji eTakahashi

    2014-06-01

    Full Text Available The transfer of extracellular RNA is emerging as an important mechanism for intracellular communication. The ability for the transfer of functionally active RNA molecules from one cell to another within vesicles such as exosomes enables a cell to modulate cellular signaling and biological processes within recipient cells. The study of extracellular RNA requires sensitive methods for the detection of these molecules. In this methods article, we will describe protocols for the detection of such extracellular RNA using sensitive detection technologies such as digital PCR. These protocols should be valuable to researchers interested in the role and contribution of extracellular RNA to tumor cell biology.

  5. Estudio molecular mediante reacción en cadena de la polimerasa y análisis de heteroduplex para el gen de la resistencia a la Rifampicina en Micobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    P. Salcedo

    2001-07-01

    Full Text Available El Micobacterium tuberculosis, es un bacilo intracelular gramnegativo, facultativo, no esporulado, aerobio estricto, de crecimiento lento y que requiere medio de cultivo complejo para su aislamiento y caracterización. La tuberculosis es una infección que cursa con diversas manifestaciones clínicas teniendo como órgano blanco el pulmón y amplia distribució mundial. El bacilo muestra resistencia o multiresistencia a los tratamientos convencionales. El objetivo del presente trabajo, mediante PCR y el análisis de Heteroduplex de una región del gen rpoB de M. tuberculosis, es detectar las cepas resistentes a la rifampicina.

  6. Determining Fungi rRNA Copy Number by PCR

    Science.gov (United States)

    The goal of this project is to improve the quantification of indoor fungal pollutants via the specific application of quantitative PCR (qPCR). Improvement will be made in the controls used in current qPCR applications. This work focuses on the use of two separate controls within ...

  7. Real-Time PCR for Universal Phytoplasma Detection and Quantification

    DEFF Research Database (Denmark)

    Christensen, Nynne Meyn; Nyskjold, Henriette; Nicolaisen, Mogens

    2013-01-01

    Currently, the most efficient detection and precise quantification of phytoplasmas is by real-time PCR. Compared to nested PCR, this method is less sensitive to contamination and is less work intensive. Therefore, a universal real-time PCR method will be valuable in screening programs and in other...

  8. Comparison of a Commercially Available Repetitive-Element PCR System (DiversiLab) with PCR Ribotyping for Typing of Clostridium difficile Strains ▿

    OpenAIRE

    Eckert, C.; Van Broeck, J.; Spigaglia, P.; Burghoffer, B.; Delmée, M.; Mastrantonio, P.; Barbut, F.

    2011-01-01

    This study compared a repetitive-element PCR (rep-PCR) method (DiversiLab system) to PCR ribotyping. The discriminatory power of rep-PCR was 0.997. Among the PCR ribotype 027 isolates tested, different rep types could be distinguished. rep-PCR showed a higher discriminatory power than PCR ribotyping. Nevertheless, this method requires technical skill, and visual interpretation of rep-PCR fingerprint patterns may be difficult.

  9. Automated PCR setup for forensic casework samples using the Normalization Wizard and PCR Setup robotic methods.

    Science.gov (United States)

    Greenspoon, S A; Sykes, K L V; Ban, J D; Pollard, A; Baisden, M; Farr, M; Graham, N; Collins, B L; Green, M M; Christenson, C C

    2006-12-20

    Human genome, pharmaceutical and research laboratories have long enjoyed the application of robotics to performing repetitive laboratory tasks. However, the utilization of robotics in forensic laboratories for processing casework samples is relatively new and poses particular challenges. Since the quantity and quality (a mixture versus a single source sample, the level of degradation, the presence of PCR inhibitors) of the DNA contained within a casework sample is unknown, particular attention must be paid to procedural susceptibility to contamination, as well as DNA yield, especially as it pertains to samples with little biological material. The Virginia Department of Forensic Science (VDFS) has successfully automated forensic casework DNA extraction utilizing the DNA IQ(trade mark) System in conjunction with the Biomek 2000 Automation Workstation. Human DNA quantitation is also performed in a near complete automated fashion utilizing the AluQuant Human DNA Quantitation System and the Biomek 2000 Automation Workstation. Recently, the PCR setup for casework samples has been automated, employing the Biomek 2000 Automation Workstation and Normalization Wizard, Genetic Identity version, which utilizes the quantitation data, imported into the software, to create a customized automated method for DNA dilution, unique to that plate of DNA samples. The PCR Setup software method, used in conjunction with the Normalization Wizard method and written for the Biomek 2000, functions to mix the diluted DNA samples, transfer the PCR master mix, and transfer the diluted DNA samples to PCR amplification tubes. Once the process is complete, the DNA extracts, still on the deck of the robot in PCR amplification strip tubes, are transferred to pre-labeled 1.5 mL tubes for long-term storage using an automated method. The automation of these steps in the process of forensic DNA casework analysis has been accomplished by performing extensive optimization, validation and testing of the

  10. Comparison of nested PCR and qPCR for the detection and quantitation of BoHV6 DNA.

    Science.gov (United States)

    Kubiś, Piotr; Materniak, Magdalena; Kuźmak, Jacek

    2013-12-01

    Nested PCR and qPCR (quantitative PCR) tests based on glycoprotein B (gB) gene were designed for detecting Bovine herpesvirus 6 (BoHV6) in bovine whole blood samples and wild ruminant blood clots (deer and roe-deer). This virus, commonly known as BLHV (bovine lymphotropic herpesvirus) belongs to the Herpesviridae family, subfamily Gammaherpesvirinae and Macavirus genus. DNA isolated from 92 dairy cow blood samples and 69 wild ruminant clots were examined for the presence of BoHV6 using nested PCR and qPCR tests. Viral DNA was detected by using nested PCR in 59 out of 92 bovine blood samples (64.1%), and by qPCR in 68 out of 92 bovine blood samples (73.9%), but none out of 69 DNA samples isolated from wild ruminant blood clots, was positive in both assays. The specificity of nested PCR and qPCR was confirmed by using BoHV1, BoHV4, BoHV6, BFV, BIV, and BLV DNA. The sensitivity of nested PCR and qPCR was determined using a serially 10-fold diluted vector pCR2.1HgB (2 × 10(0)-2 × 10(6)copies/reaction). In this testing, qPCR was more sensitive than the nested PCR, detecting two copies of BoHV6 whilst the limit of detection for nested PCR was 20 copies. In all qPCR assays, the coefficients of determination (R(2)) ranged between 0.990 and 0.999, and the calculated amplification efficiencies (Eff%) within the range of 89.7-106.9. The intra- and inter-assay CV (coefficient of variation) values did not exceed 4%. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Identification of periodontopathogen microorganisms by PCR technique

    Directory of Open Access Journals (Sweden)

    Milićević Radovan

    2008-01-01

    Full Text Available INTRODUCTION Periodontitis is an inflammatory disease of the supporting tissues of teeth and is a major cause of tooth loss in adults. The onset and progression of periodontal disease is attributed to the presence of elevated levels of a consortium of pathogenic bacteria. Gram negative bacteria, mainly strict anaerobes, play the major role. OBJECTIVE The present study aimed to assess the presence of the main types of microorganisms involved in the aetiopathogenesis of periodontal disease: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Treponema denticola, Tanerella forsythia and Prevotella intermedia in different samples collected from the oral cavity of 90 patients diagnosed with periodontitis. METHOD Bacterial DNA detection was performed in diverse biological materials, namely in dental plaque, gingival tissue and saliva, by means of multiplex PCR, a technique that allows simultaneous identification of two different bacterial genomes. RESULTS In the dental plaque of the periodontitis patients, Treponema denticola dominated. In the gingival tissue, Tannerella forsythia and Treponema denticola were the microbiota most frequently detected, whilst in saliva Treponema denticola and Eikenella corrodens were found with the highest percentage. CONCLUSION The identification of microorganisms by multiplex PCR is specific and sensitive. Rapid and precise assessment of different types of periodontopathogens is extremely important for early detection of the infection and consequently for the prevention and treatment of periodontal disease. In everyday clinical practice, for routine bacterial evaluation in patients with periodontal disease, the dental plaque is the most suitable biological material, because it is the richest in periodontal bacteria.

  12. Denoising PCR-amplified metagenome data

    Directory of Open Access Journals (Sweden)

    Rosen Michael J

    2012-10-01

    Full Text Available Abstract Background PCR amplification and high-throughput sequencing theoretically enable the characterization of the finest-scale diversity in natural microbial and viral populations, but each of these methods introduces random errors that are difficult to distinguish from genuine biological diversity. Several approaches have been proposed to denoise these data but lack either speed or accuracy. Results We introduce a new denoising algorithm that we call DADA (Divisive Amplicon Denoising Algorithm. Without training data, DADA infers both the sample genotypes and error parameters that produced a metagenome data set. We demonstrate performance on control data sequenced on Roche’s 454 platform, and compare the results to the most accurate denoising software currently available, AmpliconNoise. Conclusions DADA is more accurate and over an order of magnitude faster than AmpliconNoise. It eliminates the need for training data to establish error parameters, fully utilizes sequence-abundance information, and enables inclusion of context-dependent PCR error rates. It should be readily extensible to other sequencing platforms such as Illumina.

  13. Gluteoplastia tridimensional mediante distribución volumétrica precisa

    Directory of Open Access Journals (Sweden)

    R. Alfonso Vallarta-Rodríguez

    Full Text Available Introducción y objetivo. La gluteoplastia mediante lipoinyección debe ser una cirugía segura que partiendo de una planificación adecuada, permita un aumento moderado enfatizando contornos y mejorando la forma natural de la región glútea. Debe permitir obtener resultados predecibles, duraderos y reproducibles, además de ser aplicable en una amplia variedad de pacientes. Presentamos un método de gluteoplastia de aumento sistematizada con lipoinyección que además de ser reproducible, permite obtener resultados consistentes, naturales y permanentes, distribuyendo estratégicamente volúmenes en cuadrantes. Pacientes y Método. Con mínima manipulación del lipoaspirado, infiltramos cantidades controladas en 9 cuadrantes en cada nalga. El cuadrante central representa la zona de máxima proyección y recibe la mitad del volumen. Denominamos zonas primarias a los 4 cuadrantes en los ejes X-Y, zonas que reciben el 40% del volumen infiltrado. Las zonas secundarias o menores corresponden a los cuadrantes situados entre los cuadrantes principales, y reciben el 10% del volumen total. Resultados. Entre 2008 y 2013 intervenimos a 75 pacientes para aumento y remodelación de glúteos con la técnica descrita, todas mujeres de 24 a 52 años. Las pacientes presentaron una convalecencia favorable y una satisfacción del 93%. Nueve pacientes presentaron seromas que se resolvieron mediante aspiración en consultorio. No se presentaron complicaciones mayores. Conclusiones. Presentamos un método de remodelación glútea mediante lipoinyección que, además de ofrecer excelentes resultados, predecibles, consistentes, naturales y permanentes, es lógico y reproducible.

  14. Control de la mano robot Inmoov-SR mediante casco NeuroSky Mindset

    OpenAIRE

    Hernández Martínez, Antonio

    2017-01-01

    En este trabajo el objetivo es conseguir controlar los movimientos de apertura y cierre de la mano robot InMoov-SR conectada al brazo IRB120 de ABB mediante señales EEG, recogidas por medio del casco NeuroSky Mindset. Las señales son recogidas cuando el sujeto está en estado basal y cuando realiza movimiento con su mano y son procesadas con la ayuda de Matlab para de esta manera conseguir establecer las señales de control necesarias para activar la apertura o el cierre de la mano. Final...

  15. Protocolo de comunicación trabajador-robot mediante imágenes

    OpenAIRE

    Castilla Berduque, José Angel

    2015-01-01

    La idea del proyecto viene del concepto de “fábricas del futuro”, donde las barreras entre robots y humanos se rompen para que la colaboración entre ambos sea como en un equipo. Para la realización de este proyecto se ha utilizado el brazo robótico IRB120 de la marca ABB de 6 Grados de libertad, Matlab y el software Robot Studio. El Objetivo principal de este proyecto es establecer el protocolo de comunicación trabajador-robot mediante imágenes. El trabajador debería poder ...

  16. Estudio mediante resonancia magnética de efectos pretransicionales en cristales líquidos

    OpenAIRE

    Vaca Chavez, Fabián

    2002-01-01

    Tesis (Doctor en Física)--Universidad Nacional de Córdoba. Facultad de Matemática, Astronomía y Física, 2002. Se presenta el estudio mediante la técnica de Resonancia Magnética Nuclear de los efectos pretransicionales en diferentes fases de cristales líquidos termotrópicos y liotrópicos. Estos compuestos son materia de innumerables trabajos tanto teóricos como experimentales, debido a que son materiales extremadamente interesantes por sus aplicaciones tecnológicas, ópticas y biológicas. Se...

  17. Clasificación automática mediante la CDU con el procedimiento en cadena

    OpenAIRE

    San Segundo Manuel, Rosa

    2002-01-01

    Actas de las I Jornadas de Tratamiento y Recuperación de Información (JOTRI), Valencia, España, 4-5 julio 2002 Se entiende por clasificación automática el proceso de agrupar según el contenido las referencias de los documentos o bien los propios documentos electróneos. Este proceso se realiza mediante programas capaces de comparar términos empleados utilizados en el documento. E incluso hay otras formas automáticas de clasificación que emplean procedimientos auto...

  18. Evaluacion de competencias mediante prácticas dirigidas sobre proyectos de edificación

    OpenAIRE

    Castilla, Franciso; Castilla, Franciso; Sanz, David; González, Jesús; Pérez, Víctor

    2011-01-01

    La Búsqueda de herramientas eficaces para la evaluación de competencias transversales, comunes a diferentes asignaturas de un mismo plan de estudios, es uno de los pilares del nuevo Espacio Europeo de Educación Superior. El trabajo que aquí se presenta pretende mostrar las experiencias realizadas en primer curso del Grado en Ingeniería de Edificación en la Escuela Politécnica de Cuenca mediante prácticas dirigidas sobre edificios y proyectos de edificación. El objetivo principal es obtener...

  19. Control de costos mediante el análisis de valor ganado : caso aplicativo

    OpenAIRE

    Prado Ponce, Eduard Javier; Prado Ponce, Eduard Javier; Prado Ponce, Eduard Javier

    2015-01-01

    El control de costos y el control de plazos son muy importantes ya que pueden dar como resultados informes que permitan aplicar planes correctivos e incluso preventivos si se analizan con suficiente antelación. Existen empresas que se dedican a la construcción de obras civiles, que actualmente carece de un proceso para la planificación y control en la ejecución de obra, debido a ello surge la necesidad de desarrollar una metodología que permita a la empresa optimizar sus recursos mediante ...

  20. Monitorización de un lecho fluidizado mediante acelerometría

    OpenAIRE

    Velasco Fernández, Mario

    2013-01-01

    El presente proyecto estudiará la posibilidad de monitorizar un reactor químico mediante sensores de vibración. Actualmente, no se realiza este tipo de monitorización sobre reactores químicos, y los estudios realizados al respecto son escasos. Se tratarán de establecer las posibles equivalencias entre las medidas realizadas con sensores de presión y de vibración. Para ello se realizará la monitorización de un modelo de reactor a escala, del laboratorio de la Universidad, utilizand...

  1. Detección facial y reconocimiento anímico mediante las expresiones faciales

    OpenAIRE

    BARTUAL GONZÁLEZ, RAQUEL

    2017-01-01

    The project is based on the software development elaborated with the program LabView. The mentioned program aims to detect people's faces in a video, as well as their genre and mood state. El proyecto se basa en el desarrollo de un software elaborado con el programa LabView. Dicho programa pretende detectar la cara de las personas en un vídeo, así como su género y su estado de ánimo. Bartual González, R. (2017). Detección facial y reconocimiento anímico mediante las expresiones faciales...

  2. Estabilización de Suelos mediante el empleo de Sales Cuaternarias

    OpenAIRE

    Juan M. Junco del Pino

    2010-01-01

    El Mundo se dirige hacia el aprovechamiento de los Suelos mediante el desarrollo de nuevas técnicas y adaptarse a las condiciones del entorno resulta importante para la Ingeniería. El mejoramiento de los suelos abre nuevas posibilidades de ahorro que pueden llegar de 20 a 45 % respecto a los costos de construcción convencional. La Estabilización Química de Suelos consiste en el empleo de sustancias químicas con el objetivo de modificar las propiedades del suelo para hacerlo más denso o increm...

  3. Metalgoritmo de optimización combinatoria mediante la exploración de grafos.

    OpenAIRE

    Pastor, Rafael

    1999-01-01

    Actualmente, aunque existen procedimientos específicos para resolver de forma óptima algunos problemas concretos de optimización combinatoria, la mayoría se deben solucionar con técnicas generales de exploración del espacio de soluciones, y más concretamente mediante procedimientos de exploración enumerativos en árboles y grafos de búsqueda.Se analizan los procedimientos de este tipo expuestos en la literatura, tanto en el área de la investigación operativa como en el de la inteligencia artif...

  4. Ensayo no destructivo de soldaduras en pernos conectores mediante inspección acústica

    Directory of Open Access Journals (Sweden)

    Aznar, A.

    2012-09-01

    Full Text Available Headed studs are nowadays the standard steel-concrete connectors because of their competitive advantages. Firstly, they provide a high degree of safety thanks to semiautomatic electric arc welding. These welds are not suitable for typical non-destructive tests. The analytical study comprises several models. The first vibration modes have been obtained. The experimental research has developed first the measurement of the natural frequencies of 28 headed-studs in the sonic range. Then they have been tested by non-destructive and destructive tests. Finally theirs tests have been compared with their respective frequency measurements. A clear relationship between the measured frequencies and the lack of penetration of the welds has been established, that confirms the analytical prediction of this effect of the internal weld imperfections. Therefore, the feasibility of simple and absolutely non-destructive tests of welded studs by in site measurement of natural frequencies in the sonic range has been clearly established in this work.

    Los pernos conectores aportan múltiples ventajas de uso, entre las que se encuentra el elevado margen de seguridad que ofrecen sus soldaduras ejecutadas mediante arco eléctrico. Estas soldaduras, aunque ampliamente fiables, son difícilmente comprobadas mediante ensayos no destructivos. El presente estudio plantea la inspección de soldaduras de pernos conectores mediante su espectro acústico. Analíticamente, la investigación se ha centrado en el cálculo de los primeros modos propios de vibración. Experimentalmente se han medido las frecuencias propias de resonancia de 28 pernos, en los que posteriormente se han llevado a cabo ensayos tanto no destructivos como destructivos. Se ha obtenido, tanto teórica como experimentalmente, una relación entre la frecuencia de vibración de los pernos conectores y la calidad de la soldadura. Por ello se verifica la posibilidad de inspección de estas

  5. Tratamiento de un efluente textil mediante electrooxidación-Salix babylonica

    OpenAIRE

    Sánchez Sánchez, Hilda Alejandra

    2016-01-01

    A nivel mundial, la industria textil es considerada una de las principales fuentes de descarga que afectan la calidad del agua debido al gran volumen que emplea en sus procesos y al uso de una amplia gama de colorantes sintéticos. En esta investigación se evaluó el tratamiento de un agua residual textil mediante un sistema acoplado de electrooxidación-Salix babylonica usando electrodos DDB. En el estudio, se construyó una celda electroquímica en batch, utilizando 5 electrodos paralelos vertic...

  6. Accurate quantification of supercoiled DNA by digital PCR

    Science.gov (United States)

    Dong, Lianhua; Yoo, Hee-Bong; Wang, Jing; Park, Sang-Ryoul

    2016-01-01

    Digital PCR (dPCR) as an enumeration-based quantification method is capable of quantifying the DNA copy number without the help of standards. However, it can generate false results when the PCR conditions are not optimized. A recent international comparison (CCQM P154) showed that most laboratories significantly underestimated the concentration of supercoiled plasmid DNA by dPCR. Mostly, supercoiled DNAs are linearized before dPCR to avoid such underestimations. The present study was conducted to overcome this problem. In the bilateral comparison, the National Institute of Metrology, China (NIM) optimized and applied dPCR for supercoiled DNA determination, whereas Korea Research Institute of Standards and Science (KRISS) prepared the unknown samples and quantified them by flow cytometry. In this study, several factors like selection of the PCR master mix, the fluorescent label, and the position of the primers were evaluated for quantifying supercoiled DNA by dPCR. This work confirmed that a 16S PCR master mix avoided poor amplification of the supercoiled DNA, whereas HEX labels on dPCR probe resulted in robust amplification curves. Optimizing the dPCR assay based on these two observations resulted in accurate quantification of supercoiled DNA without preanalytical linearization. This result was validated in close agreement (101~113%) with the result from flow cytometry. PMID:27063649

  7. A survey of tools for the analysis of quantitative PCR (qPCR data

    Directory of Open Access Journals (Sweden)

    Stephan Pabinger

    2014-09-01

    Our comprehensive survey showed that most tools use their own file format and only a fraction of the currently existing tools support the standardized data exchange format RDML. To allow a more streamlined and comparable analysis of qPCR data, more vendors and tools need to adapt the standardized format to encourage the exchange of data between instrument software, analysis tools, and researchers.

  8. Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR

    Directory of Open Access Journals (Sweden)

    Roman Wölfel

    2012-08-01

    Full Text Available Ricin and abrin are two of the most potent plant toxins known and may be easily obtained in high yield from the seeds using rather simple technology. As a result, both toxins are potent and available toxins for criminal or terrorist acts. However, as the production of highly purified ricin or abrin requires sophisticated equipment and knowledge, it may be more likely that crude extracts would be used by non-governmental perpetrators. Remaining plant-specific nucleic acids in these extracts allow the application of a real-time PCR (qPCR assay for the detection and identification of abrin or ricin genomic material. Therefore, we have developed a duplex real-time PCR assays for simultaneous detection of ricin and abrin DNA based on the OmniMix HS bead PCR reagent mixture. Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5′-nuclease technology. The assay was thoroughly optimized and validated in terms of analytical sensitivity. Evaluation of the assay sensitivity by probit analysis demonstrated a 95% probability of detection at 3 genomes per reaction for ricin DNA and 1.2 genomes per reaction for abrin DNA. The suitability of the assays was exemplified by detection of ricin and abrin contaminations in a food matrix.

  9. Analytical Performance of Four Polymerase Chain Reaction (PCR and Real Time PCR (qPCR Assays for the Detection of Six Leishmania Species DNA in Colombia

    Directory of Open Access Journals (Sweden)

    Cielo M. León

    2017-10-01

    Full Text Available Leishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR, limit of detection (LoD and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia. Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America.

  10. Analytical Performance of Four Polymerase Chain Reaction (PCR) and Real Time PCR (qPCR) Assays for the Detection of Six Leishmania Species DNA in Colombia

    Science.gov (United States)

    León, Cielo M.; Muñoz, Marina; Hernández, Carolina; Ayala, Martha S.; Flórez, Carolina; Teherán, Aníbal; Cubides, Juan R.; Ramírez, Juan D.

    2017-01-01

    Leishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR) including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets) and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR), limit of detection (LoD) and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively) and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia). Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America. PMID:29046670

  11. DNA polymerase preference determines PCR priming efficiency.

    Science.gov (United States)

    Pan, Wenjing; Byrne-Steele, Miranda; Wang, Chunlin; Lu, Stanley; Clemmons, Scott; Zahorchak, Robert J; Han, Jian

    2014-01-30

    Polymerase chain reaction (PCR) is one of the most important developments in modern biotechnology. However, PCR is known to introduce biases, especially during multiplex reactions. Recent studies have implicated the DNA polymerase as the primary source of bias, particularly initiation of polymerization on the template strand. In our study, amplification from a synthetic library containing a 12 nucleotide random portion was used to provide an in-depth characterization of DNA polymerase priming bias. The synthetic library was amplified with three commercially available DNA polymerases using an anchored primer with a random 3' hexamer end. After normalization, the next generation sequencing (NGS) results of the amplified libraries were directly compared to the unamplified synthetic library. Here, high throughput sequencing was used to systematically demonstrate and characterize DNA polymerase priming bias. We demonstrate that certain sequence motifs are preferred over others as primers where the six nucleotide sequences at the 3' end of the primer, as well as the sequences four base pairs downstream of the priming site, may influence priming efficiencies. DNA polymerases in the same family from two different commercial vendors prefer similar motifs, while another commercially available enzyme from a different DNA polymerase family prefers different motifs. Furthermore, the preferred priming motifs are GC-rich. The DNA polymerase preference for certain sequence motifs was verified by amplification from single-primer templates. We incorporated the observed DNA polymerase preference into a primer-design program that guides the placement of the primer to an optimal location on the template. DNA polymerase priming bias was characterized using a synthetic library amplification system and NGS. The characterization of DNA polymerase priming bias was then utilized to guide the primer-design process and demonstrate varying amplification efficiencies among three commercially

  12. Molecular quantification of environmental DNA using microfluidics and digital PCR.

    Science.gov (United States)

    Hoshino, Tatsuhiko; Inagaki, Fumio

    2012-09-01

    Real-time PCR has been widely used to evaluate gene abundance in natural microbial habitats. However, PCR-inhibitory substances often reduce the efficiency of PCR, leading to the underestimation of target gene copy numbers. Digital PCR using microfluidics is a new approach that allows absolute quantification of DNA molecules. In this study, digital PCR was applied to environmental samples, and the effect of PCR inhibitors on DNA quantification was tested. In the control experiment using λ DNA and humic acids, underestimation of λ DNA at 1/4400 of the theoretical value was observed with 6.58 ng μL(-1) humic acids. In contrast, digital PCR provided accurate quantification data with a concentration of humic acids up to 9.34 ng μL(-1). The inhibitory effect of paddy field soil extract on quantification of the archaeal 16S rRNA gene was also tested. By diluting the DNA extract, quantified copy numbers from real-time PCR and digital PCR became similar, indicating that dilution was a useful way to remedy PCR inhibition. The dilution strategy was, however, not applicable to all natural environmental samples. For example, when marine subsurface sediment samples were tested the copy number of archaeal 16S rRNA genes was 1.04×10(3) copies/g-sediment by digital PCR, whereas real-time PCR only resulted in 4.64×10(2) copies/g-sediment, which was most likely due to an inhibitory effect. The data from this study demonstrated that inhibitory substances had little effect on DNA quantification using microfluidics and digital PCR, and showed the great advantages of digital PCR in accurate quantifications of DNA extracted from various microbial habitats. Copyright © 2012 Elsevier GmbH. All rights reserved.

  13. Multiplex Amplification Refractory Mutation System PCR (ARMS-PCR) provides sequencing independent typing of canine parvovirus.

    Science.gov (United States)

    Chander, Vishal; Chakravarti, Soumendu; Gupta, Vikas; Nandi, Sukdeb; Singh, Mithilesh; Badasara, Surendra Kumar; Sharma, Chhavi; Mittal, Mitesh; Dandapat, S; Gupta, V K

    2016-12-01

    Canine parvovirus-2 antigenic variants (CPV-2a, CPV-2b and CPV-2c) ubiquitously distributed worldwide in canine population causes severe fatal gastroenteritis. Antigenic typing of CPV-2 remains a prime focus of research groups worldwide in understanding the disease epidemiology and virus evolution. The present study was thus envisioned to provide a simple sequencing independent, rapid, robust, specific, user-friendly technique for detecting and typing of presently circulating CPV-2 antigenic variants. ARMS-PCR strategy was employed using specific primers for CPV-2a, CPV-2b and CPV-2c to differentiate these antigenic types. ARMS-PCR was initially optimized with reference positive controls in two steps; where first reaction was used to differentiate CPV-2a from CPV-2b/CPV-2c. The second reaction was carried out with CPV-2c specific primers to confirm the presence of CPV-2c. Initial validation of the ARMS-PCR was carried out with 24 sequenced samples and the results were matched with the sequencing results. ARMS-PCR technique was further used to screen and type 90 suspected clinical samples. Randomly selected 15 suspected clinical samples that were typed with this technique were sequenced. The results of ARMS-PCR and the sequencing matched exactly with each other. The developed technique has a potential to become a sequencing independent method for simultaneous detection and typing of CPV-2 antigenic variants in veterinary disease diagnostic laboratories globally. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. IMPLEMENTACIÓN DE LA PCR (REACCIÓN EN CADENA DE LA POLIMERASA PARA EL DIAGNÓSTICO DE LA BRUCELOSIS DE BOVINOS EN EL ECUADOR

    Directory of Open Access Journals (Sweden)

    Orly Fernando Cevallos Falquez

    2008-06-01

    Full Text Available En el presente trabajo se implementó la técnica de PCR para la detección de Brucella abortus, en muestras de sangre, en comparación con la prueba Rosa de bengala. La amplificación del ADN se realizó utilizando tres oligonucleotidos homólogos correspondientes a la secuencia 16 ARNr de Brucella abortus, dando una amplificación de 900 y 725 pb respectivamente. Un total de 172 muestras de sangre fueron recolectadas de 4 hatos con prevalencia histórica de presencia de brucelosis. Resultaron 142 negativas con la prueba de serológia y 143 con PCR, 30 resultaron positivas con la prueba serológica Rosa de bengala y 29 salieron positivas con PCR. Todos los animales que salieron positivos con Rosa de bengala, 4 presentaban síntomas clínicos como son; abortos, terneros débiles, baja producción de carne y leche. Los otros 26 animales resultaron negativos con PCR y estos animales no presentaron los síntomas clínicos. De los 142 animales que dieron negativo con Rosa de bengala, 25 resultaron positivos con PCR. Los resultados muestran que la detección de los animales positivos mediante la PCR fue más especificas y sensibles que la prueba serológica de Rosa de bengala, por lo tanto es una herramienta muy útil en el diagnóstico de Brucella abortus.

  15. ReadqPCR and NormqPCR: R packages for the reading, quality checking and normalisation of RT-qPCR quantification cycle (Cq data

    Directory of Open Access Journals (Sweden)

    Perkins James R

    2012-07-01

    Full Text Available Abstract Background Measuring gene transcription using real-time reverse transcription polymerase chain reaction (RT-qPCR technology is a mainstay of molecular biology. Technologies now exist to measure the abundance of many transcripts in parallel. The selection of the optimal reference gene for the normalisation of this data is a recurring problem, and several algorithms have been developed in order to solve it. So far nothing in R exists to unite these methods, together with other functions to read in and normalise the data using the chosen reference gene(s. Results We have developed two R/Bioconductor packages, ReadqPCR and NormqPCR, intended for a user with some experience with high-throughput data analysis using R, who wishes to use R to analyse RT-qPCR data. We illustrate their potential use in a workflow analysing a generic RT-qPCR experiment, and apply this to a real dataset. Packages are available from http://www.bioconductor.org/packages/release/bioc/html/ReadqPCR.htmland http://www.bioconductor.org/packages/release/bioc/html/NormqPCR.html Conclusions These packages increase the repetoire of RT-qPCR analysis tools available to the R user and allow them to (amongst other things read their data into R, hold it in an ExpressionSet compatible R object, choose appropriate reference genes, normalise the data and look for differential expression between samples.

  16. Monitorización de Signos Vitales Mediante una Red de Dispositivos Móviles

    Directory of Open Access Journals (Sweden)

    Daniel Cilio

    2013-11-01

    Full Text Available El desarrollo e implementación de diferentes proyectos tecnológicos, apoyados en el correspondiente conocimiento médico, pueden contribuir a resolver varios problemas del sector de la salud. Si bien en los últimos años se han realizado enormes esfuerzos para desarrollar tecnologías aplicables en ambientes clínicos, el desarrollo de tecnologías para atención médica domiciliar podría reducir la presión que agobia a los hospitales actualmente. En el presente proyecto se realiza el diseño e implementación de un sistema para monitorización de signos vitales, el cual mide la frecuencia cardiaca, la oxigenación sanguínea y la temperatura corporal de una persona. La información obtenida de cada signo vital es muestreada y procesada por una plataforma digital para posteriormente ser enviada mediante un módulo Bluetooth hacia un dispositivo móvil para su análisis y visualización. El prototipo fue evaluado mediante una batería de pruebas para medición de signos vitales en diferentes pacientes.

  17. Pronósticos de inflación mediante técnicas bayesianas

    Directory of Open Access Journals (Sweden)

    Juan Diego Chavarría

    2015-11-01

    Full Text Available La efectividad de la política monetaria bajo un esquema de metas de inflación como el propuesto por el Banco Central de Costa Rica se basa en buena medida en el correcto y oportuno pronóstico de la inflación a corto y mediano plazo con el fin de diseñar de mejor forma las acciones de política monetaria. Así, el propósito de este trabajo es desarrollar una herramienta complementaria para elaborar pronósticos de inflación mediante un enfoque bayesiano. Para lo anterior se propone la utilización de la metodología Bayesian Model Averaging y de Weighted Average Least Squares. Los modelos de proyección especificados permitirían ampliar y complementar el análisis que se realiza actualmente con el Modelo Macroeconómico de Proyección Trimestral (MMPT del Banco Central de Costa Rica. Como resultado esta investigación muestra que, para datos de periodicidad mensual y a horizontes de pronóstico de 1 a 12 meses, es posible encontrar proyecciones mediante un proceso bayesiano que poseen una mayor capacidad predictiva en relación con aquellas producidas por un modelo autorregresivo.

  18. Principios básicos y aplicación del aprendizaje mediante tareas

    Directory of Open Access Journals (Sweden)

    Estaire, Sheila

    2011-04-01

    Full Text Available En los últimos años el aprendizaje mediante tareas ha ido consolidándose como una nueva forma de enseñar y aprender lenguas extranjeras. Sin embargo existen una serie de aspectos prácticos relacionados con su aplicación en los que aún se puede profundizar. En este artículo, después de una introducción breve de algunos principios básicos, se discuten posibles procedimientos para determinar las tareas que constituyen el eje de un programa, así como para organizar el proceso de enseñanza / aprendizaje. A continuación se presentan diferentes modalidades de trabajo sobre los aspectos formales de la lengua, aspectos que es esencial tratar de forma rigurosa, minuciosa y sistemática. Este punto crucial se discute junto con una propuesta de estructura de curso que consta de dos componentes diferenciados. Por otra parte, los elementos innovadores del aprendizaje mediante tareas hacen imprescindible una gestión del aprendizaje también innovadora, aspecto que se trata en el último apartado a través de pautas metodológicas que potencian la eficacia de las tareas como instrumento de aprendizaje.

  19. Ahorro energético mediante estrategias de iluminación natural optimizadas

    Directory of Open Access Journals (Sweden)

    Puigdomènech Franquesa, Joan

    1986-04-01

    Full Text Available Electrical charges in buildings and specially in those of commercial use, can be diminished by means of natural lighting strategies. Taking the climate features of our country into consideration, it is necessary to prevent the inconveniences caused by an en erg y excess in summer, so solar Controls are needed. The only practical way to achieve the suitable balance between thermal and light needs, so as to get a monthly or annual energetic balance optimization, is to operate with the computer. A programme with such characteristics is described here. Its application gives important sarings in non renouvable energy savings.Mediante estrategias de iluminación natural es posible disminuir las cargas eléctricas de los edificios y en especial los de uso comercial. Dadas las características climáticas de nuestro país es necesario prever los inconvenientes de un exceso de energía en verano, para lo cual es preciso disponer de controles solares. Encontrar el correcto equilibrio entre las necesidades térmicas y lumínicas en base a la optimización del balance energético mensual o anual es únicamente factible mediante el uso del ordenador. Un programa que responde a estas características es descrito en el presente trabajo, obteniéndose con su aplicación importantes ahorros en el consumo de energías no renovables.

  20. PROPUESTA DE CONEXIÓN DE ENTORNOS IPv6 MEDIANTE UN BACKBONE MPLS/IPv4

    Directory of Open Access Journals (Sweden)

    Nancy Yaneth Gelvez García

    2013-09-01

    Full Text Available Las redes actuales MPLS/IPv4 presentan las ventajas de poder implementar ingeniería de tráfico, así como realizar diferenciación de flujos mediante clases de servicio (CoS frente a las redes con enrutamiento IP tradicional. En aras de aprovechar cualidades estratégicas durante la etapa de coexistencia entre IPv4 e IPv6 existen 4 métodos para proveer conectividad a islas IPv6 [1] remotas a través de una infraestructura de core MPLS con IPv4 nativo [2], sin embargo una de las formas que permite un rápida y fácil provisión de la misma dados los mínimos requisitos de configuración y de equipos es la de disponer túneles IPv6 en los enrutadores de acceso (CE de la red. No obstante, sus cuatro variantes (manual, GRE, 6to4 e IPv6 compatible IPv4 [3] resultan adecuadas o no según las características inherentes de la red a interconectar; por tanto este artículo presenta las ventajas y desventajas propias de la utilización de cada técnica de entunelamiento como resultado de la interconexión con los cuatro tipos de túneles de una red emulada mediante GNS3+Dynamips.

  1. Modelo de dinámica lateral de vehículo mediante bond graph

    Directory of Open Access Journals (Sweden)

    Juan Carlos Parra Márquez

    2008-07-01

    Full Text Available Este trabajo presenta los resultados de la investigación, cuyo objetivo es obtener un modelo matemático que permita determinar la dinámica lateral de un vehículo mediante el uso de Bond Graph. Este modelo es válido para robótica móvil. Los análisis de comportamiento del modelo han sido probados con simulaciones típicas del movimiento lateral de un vehículo. Finalmente, este modelo ha sido obtenido e implementado mediante el software 20-Sim. This paper presents the results of a research whose objective was to find a mathematical model in order to determine the lateral dynamic of Vehicle by means of the use of Bond Graph. This model is valid also for mobile robotics. The analyses of behavior of the model were realized across typical simulations of a vehicle in lateral movement. Finally, this mathematical model was obtained and implemented across the software 20-Sim.

  2. DIMENSIONAMIENTO DE UN SISTEMA DE ENERGÍA TERMOSOLAR MEDIANTE EL USO DE UN MODELO

    Directory of Open Access Journals (Sweden)

    Arturo Daniel Alarcón Rodríguez

    2004-01-01

    Full Text Available En el presente artículo se expone el método de dimensionamiento de sistemas termosolares mediante el uso de un modelo matemático. Este método es comúnmente usado debido que es simple, flexible pero a la vez muy potente. La simulación del sistema termosolar se realiza en base a un modelo matemático que describe los fenómenos térmicos que ocurren mediante un conjunto de ecuaciones diferenciales. Los parámetros que determinan el modelo son coeficientes de intercambio de calor entre los elementos del sistema, parámetros que representan las características de los componentes del sistema termosolar y parámetros que representan las condiciones en las que trabajará el sistema. Estos parámetros se determinan en base a recomendaciones de bibliografía, observaciones, mediciones de campo y correlaciones adecuadas. El uso de un modelo para el dimensionamiento de un sistema termosolar resulta una herramienta muy útil, ya que se adapta a distintas configuraciones de sistemas termosolares. Permite asimismo, tener una idea bastante aproximada del comportamiento del sistema termosolar en distintas condiciones de uso, la que sólo podría obtenerse a través de experimentos físicos complicados y por ende costosos.

  3. Digital PCR as a tool to measure HIV persistence.

    Science.gov (United States)

    Rutsaert, Sofie; Bosman, Kobus; Trypsteen, Wim; Nijhuis, Monique; Vandekerckhove, Linos

    2018-01-30

    Although antiretroviral therapy is able to suppress HIV replication in infected patients, the virus persists and rebounds when treatment is stopped. In order to find a cure that can eradicate the latent reservoir, one must be able to quantify the persisting virus. Traditionally, HIV persistence studies have used real-time PCR (qPCR) to measure the viral reservoir represented by HIV DNA and RNA. Most recently, digital PCR is gaining popularity as a novel approach to nucleic acid quantification as it allows for absolute target quantification. Various commercial digital PCR platforms are nowadays available that implement the principle of digital PCR, of which Bio-Rad's QX200 ddPCR is currently the most used platform in HIV research. Quantification of HIV by digital PCR is proving to be a valuable improvement over qPCR as it is argued to have a higher robustness to mismatches between the primers-probe set and heterogeneous HIV, and forfeits the need for a standard curve, both of which are known to complicate reliable quantification. However, currently available digital PCR platforms occasionally struggle with unexplained false-positive partitions, and reliable segregation between positive and negative droplets remains disputed. Future developments and advancements of the digital PCR technology are promising to aid in the accurate quantification and characterization of the persistent HIV reservoir.

  4. Comparación del gasto energético en reposo determinado mediante calorimetría indirecta y estimado mediante fórmulas predictivas en mujeres con grados de obesidad I a III

    OpenAIRE

    Alicia Parra-Carriedo; Loren Cherem-Cherem; Daniela Galindo-De Noriega; Mary Carmen Díaz-Gutiérrez; Ana Bertha Pérez-Lizaur; César Hernández-Guerrero

    2013-01-01

    Introducción: La determinación del gasto energético en reposo (GER) se calcula cotidianamente a partir de fórmulas predictivas aunque el resultado varía dependiendo de la población. Objetivo: Comparar la determinación del GER mediante calorimetría indirecta y mediante las ecuaciones Harris-Benedict (HB), Mifflin (MF), Organización Mundial de la Salud (OMS), "Institute of Medicine" (IOM), Fórmula Rápida (FR) y Valencia (VA) en mujeres con grados de obesidad I a III. Métodos: Mujeres adultas me...

  5. COMPARISON OF 16S rRNA-PCR-RFLP, LipL32-PCR AND OmpL1-PCR METHODS IN THE DIAGNOSIS OF LEPTOSPIROSIS

    Directory of Open Access Journals (Sweden)

    Tülin GÜVEN GÖKMEN

    Full Text Available SUMMARY Leptospirosis is still one of the most important health problems in developing countries located in humid tropical and subtropical regions. Human infections are generally caused by exposure to water, soil or food contaminated with the urine of infected wild and domestic animals such as rodents and dogs. The clinical course of leptospirosis is variable and may be difficult to distinguish from many other infectious diseases. The dark-field microscopy (DFM, serology and nucleic acid amplification techniques are used to diagnose leptospirosis, however, a distinctive standard reference method is still lacking. Therefore, in this study, we aimed to determine the presence of Leptospira spp., to differentiate the pathogenic L. interrogans and the non-pathogenic L. biflexa, and also to determine the sensitivity and specificity values of molecular methods as an alternative to conventional ones. A total of 133 serum samples, from 47 humans and 86 cattle were evaluated by two conventional tests: the Microagglutination Test (MAT and the DFM, as well as three molecular methods, the 16S rRNA-PCR followed by Restriction Fragment Lenght Polymorphism (RFLP of the amplification products 16S rRNA-PCR-RFLP, LipL32-PCR and OmpL1-PCR. In this study, for L. interrogans, the specificity and sensitivity rates of the 16S rRNA-PCR and the LipL32-PCR were considered similar (100% versus 98.25% and 100% versus 98.68%, respectively. The OmpL1-PCR was able to classify L. interrogans into two intergroups, but this PCR was less sensitive (87.01% than the other two PCR methods. The 16S rRNA-PCR-RFLP could detect L. biflexa DNA, but LipL32-PCR and OmpL1-PCR could not. The 16S rRNA-PCR-RFLP provided an early and accurate diagnosis and was able to distinguish pathogenic and non-pathogenic Leptospira species, hence it may be used as an alternative method to the conventional gold standard techniques for the rapid disgnosis of leptospirosis.

  6. Multiplex enrichment quantitative PCR (ME-qPCR): a high-throughput, highly sensitive detection method for GMO identification.

    Science.gov (United States)

    Fu, Wei; Zhu, Pengyu; Wei, Shuang; Zhixin, Du; Wang, Chenguang; Wu, Xiyang; Li, Feiwu; Zhu, Shuifang

    2017-04-01

    Among all of the high-throughput detection methods, PCR-based methodologies are regarded as the most cost-efficient and feasible methodologies compared with the next-generation sequencing or ChIP-based methods. However, the PCR-based methods can only achieve multiplex detection up to 15-plex due to limitations imposed by the multiplex primer interactions. The detection throughput cannot meet the demands of high-throughput detection, such as SNP or gene expression analysis. Therefore, in our study, we have developed a new high-throughput PCR-based detection method, multiplex enrichment quantitative PCR (ME-qPCR), which is a combination of qPCR and nested PCR. The GMO content detection results in our study showed that ME-qPCR could achieve high-throughput detection up to 26-plex. Compared to the original qPCR, the Ct values of ME-qPCR were lower for the same group, which showed that ME-qPCR sensitivity is higher than the original qPCR. The absolute limit of detection for ME-qPCR could achieve levels as low as a single copy of the plant genome. Moreover, the specificity results showed that no cross-amplification occurred for irrelevant GMO events. After evaluation of all of the parameters, a practical evaluation was performed with different foods. The more stable amplification results, compared to qPCR, showed that ME-qPCR was suitable for GMO detection in foods. In conclusion, ME-qPCR achieved sensitive, high-throughput GMO detection in complex substrates, such as crops or food samples. In the future, ME-qPCR-based GMO content identification may positively impact SNP analysis or multiplex gene expression of food or agricultural samples. Graphical abstract For the first-step amplification, four primers (A, B, C, and D) have been added into the reaction volume. In this manner, four kinds of amplicons have been generated. All of these four amplicons could be regarded as the target of second-step PCR. For the second-step amplification, three parallels have been taken for

  7. Blood grouping based on PCR methods and agarose gel electrophoresis.

    Science.gov (United States)

    Sell, Ana Maria; Visentainer, Jeane Eliete Laguila

    2015-01-01

    The study of erythrocyte antigens continues to be an intense field of research, particularly after the development of molecular testing methods. More than 300 specificities have been described by the International Society for Blood Transfusion as belonging to 33 blood group systems. The polymerase chain reaction (PCR) is a central tool for red blood cells (RBC) genotyping. PCR and agarose gel electrophoresis are low cost, easy, and versatile in vitro methods for amplifying defined target DNA (RBC polymorphic region). Multiplex-PCR, AS-PCR (Specific Allele Polymerase Chain Reaction), and RFLP-PCR (Restriction Fragment Length Polymorphism-Polymerase Chain Reaction) techniques are usually to identify RBC polymorphisms. Furthermore, it is an easy methodology to implement. This chapter describes the PCR methodology and agarose gel electrophoresis to identify the polymorphisms of the Kell, Duffy, Kidd, and MNS blood group systems.

  8. Product differentiation during continuous-flow thermal gradient PCR.

    Science.gov (United States)

    Crews, Niel; Wittwer, Carl; Palais, Robert; Gale, Bruce

    2008-06-01

    A continuous-flow PCR microfluidic device was developed in which the target DNA product can be detected and identified during its amplification. This in situ characterization potentially eliminates the requirement for further post-PCR analysis. Multiple small targets have been amplified from human genomic DNA, having sizes of 108, 122, and 134 bp. With a DNA dye in the PCR mixture, the amplification and unique melting behavior of each sample is observed from a single fluorescent image. The melting behavior of the amplifying DNA, which depends on its molecular composition, occurs spatially in the thermal gradient PCR device, and can be observed with an optical resolution of 0.1 degrees C pixel(-1). Since many PCR cycles are within the field of view of the CCD camera, melting analysis can be performed at any cycle that contains a significant quantity of amplicon, thereby eliminating the cycle-selection challenges typically associated with continuous-flow PCR microfluidics.

  9. Selecting PCR for the Diagnosis of Intestinal Parasitosis

    DEFF Research Database (Denmark)

    Hartmeyer, G. N.; Hoegh, S. V.; Skov, M. N.

    2017-01-01

    Microscopy of stool samples is a labour-intensive and inaccurate technique for detection of intestinal parasites causing diarrhoea and replacement by PCR is attractive. Almost all cases of diarrhoea induced by parasites over a nine-year period in our laboratory were due to Giardia lamblia......, Cryptosporidium species, or Entamoeba histolytica detected by microscopy. We evaluated and selected in-house singleplex real-time PCR (RT-PCR) assays for these pathogens in 99 stool samples from patients suspected of having intestinal parasitosis tested by microscopy. The strategy included a genus-specific PCR...... assay for C. parvum and C. hominis, with subsequent identification by a PCR that distinguishes between the two species. G. lamblia was detected in five and C. parvum in one out of 68 microscopy-negative samples. The performance of the in-house RT-PCR assays was compared to three commercially available...

  10. Standardization of diagnostic PCR for the detection of foodborne pathogens

    DEFF Research Database (Denmark)

    Malorny, B.; Tassios, P.T.; Radstrom, P.

    2003-01-01

    In vitro amplification of nucleic acids using the polymerase chain reaction (PCR) has become, since its discovery in the 1980s, a powerful diagnostic tool for the analysis of microbial infections as well as for the analysis of microorganisms in food samples. However, despite its potential, PCR has...... neither gained wide acceptance in routine diagnostics nor been widely incorporated in standardized methods. Lack of validation and standard protocols, as well as variable quality of reagents and equipment, influence the efficient dissemination of PCR methodology from expert research laboratories to end......-user laboratories. Moreover, the food industry understandably requires and expects officially approved standards. Recognizing this, in 1999, the European Commission approved the research project, FOOD-PCR (http://www.PCR.dk), which aims to validate and standardize the use of diagnostic PCR for the detection...

  11. Manejo sostenible y sustentable de fincas productoras mediante procesos participativos en Sáchica, Boyacá

    OpenAIRE

    Ángel Eduardo Ramírez-Amaya; Germán Gonzalo Hurtado

    2013-01-01

    Objetivo. Elaborar un proyecto de desarrollo sostenible y sustentable de fincas productoras mediante procesos participativos en el municipio de Sáchica, Boyacá. Materiales y métodos. La investigación se realizó con familias campesinas de la vereda Arrayán Alto, del municipio de Sáchica, Boyacá, mediante la metodología Investigación Acción Participativa (IAP), que se centra en la participación de las comunidades para elaborar propuestas concertadas con ellas. El trabajo se desarrolló en varias...

  12. La prueba obtenida mediante coacción y su inadmisibilidad ante la Corte Interamericana de Derechos Humanos

    OpenAIRE

    Paúl Díaz, Álvaro

    2016-01-01

    La Corte Interamericana de Derechos Humanos efectúa un amplio análisis probatorio para determinar la ocurrencia de violaciones de derechos humanos. Ella tiende a ser muy flexible con la admisión de la prueba, sin perjuicio de ello estaría obligada a excluir confesiones obtenidas mediante coacción. En relación con esto, la Corte ha hecho afirmaciones que parecen propiciar la exclusión de toda prueba obtenida mediante coerción, y dar pie a la doctrina del fruto de árbol envenenado. Este artícul...

  13. Modelado del proceso de esterilización del hospital clínico universitario de Valladolid mediante diagramas IDEF

    OpenAIRE

    Viñas del Hoyo, Víctor

    2015-01-01

    El principal objetivo de este trabajo de fin de grado es elaborar mediante diagramas IDEF, más concretamente el IDEFO, cuál sería el funcionamiento de la central de esterilización de nueva construcción del Hospital Clínico Universitario de Valladolid, mediante la gestión por procesos. Otros objetivos secundarios pero no menos importantes de este proyecto son:comprender el modelo de gestión por procesos e identificar los pasos que hay que seguir para implantarla correctamente. Ver y aprender ...

  14. Mapas de Entornos Mediante Navegacion Difusa y Sistema de Teleoperacion de una Plataforma Pioneer P3-DX

    Directory of Open Access Journals (Sweden)

    Daniel Granda

    2013-11-01

    Full Text Available El presente proyecto describe el diseno e implementacion de aplicaciones de Teleoperacion, Adquisicion de Datos, Control Difuso de Velocidad y Mapeo de Entornos en 2D, para la plataforma movil Pioneer P3-DX mediante el uso de sonares, odometrıa y software libre GNU/Linux. El proyecto brinda una guıa para utilizar los conceptos de programacion en Python, que permite crear aplicaciones de manera versatil mediante el uso de librerıas como: GTK para el desarrollo del entorno grafico, PYFUZZY para el desarrollo del controlador difuso de velocidad y OPENCV para mostrar los mapas del entorno.

  15. Relación entre nutrición, polución del aire y enfermedad cardiovascular: PCR Ultrasensible en el personal policial de tránsito del Distrito Metropolitano de Quito. Diciembre 2008

    OpenAIRE

    Collaguazo Guamán, Andrés Gabriel; Nieto Nieto, Bertha Leonila

    2010-01-01

    Las afecciones cardiovasculares son enfermedades poligenéticas, que involucran múltiples factores de riesgo. Se ha demostrado también que los tóxicos ambientales ejercerían efectos sobre la capacidad de enfermarse, incrementando el riesgo cardiovascular previamente existente y potenciando otros factores de riesgo o actuando de manera independiente. Objetivo. Estudiar la relación entre Nutrición, Polución del aire y Enfermedad Cardiovascular mediante PCR ultrasensible en el Personal Poli...

  16. Comparison of multiplex reverse transcription-PCR-enzyme ...

    African Journals Online (AJOL)

    Comparison of multiplex reverse transcription-PCR-enzyme hybridization assay with immunofluorescence techniques for the detection of four viral respiratory pathogens in pediatric community acquired pneumonia.

  17. Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A

    OpenAIRE

    Ferreira, HL; Spilki, FR; dos Santos, MMAB; de Almeida, RS; Arns, CW

    2009-01-01

    Avian metapneumovirus (AMPV) belongs to Metapneumovirus genus of Paramyxoviridae family. Virus isolation, serology, and detection of genomic RNA are used as diagnostic methods for AMPV. The aim of the present study was to compare the detection of six subgroup A AMPV isolates (AMPV/A) viral RNA by using different conventional and real time RT-PCR methods. Two new RT-PCR tests and two real time RT-PCR tests, both detecting fusion (F) gene and nucleocapsid (N) gene were compared with an establis...

  18. Representación del Conocimiento en curriculo mediante esquemas preconceptuales

    Directory of Open Access Journals (Sweden)

    Carlos Mario Zapata

    2011-09-01

    Full Text Available El concepto de currículo se torna más y más complejo en tanto aparecen nuevos estudios que lo complementan. Como consecuencia, los modelos que, gráfica o formalmente, tratan de representar el conocimiento alrededor del currículo se ocupan cada vez más de aspectos locales, de este modo le restan generalidad de comprensión. Por ello, en este artículo de investigación se realiza una revisión acerca de los diferentes enfoques del currículo a lo largo del siglo XX y de los modelos que representan este concepto. Finalmente, se propone una representación integradora de las diferentes visiones de currículo mediante los denominados esquemas preconceptuales, que consisten en diagramas para la representación del conocimiento cercanos al lenguaje natural

  19. CONTROL ROBUSTO DE UN SISTEMA MECÁNICO SIMPLE MEDIANTE UNA HERRAMIENTA GRAFICA

    Directory of Open Access Journals (Sweden)

    LUINI HURTADO CORTÉS

    2010-01-01

    Full Text Available en este artículo se presenta el diseño de un controlador robusto para un sistema masaresorteamortiguador . Con el fin de realizar un diseño simple, se tomó en cuenta únicamente la incertidumbre en los parámetros de la planta. Los cálculos del problema se realizaron con una interfaz gráfica desarrollada para el diseño de controladores robustos, disponible para la Toolbox de Control Robusto de Matlab Ò . Se pretende que este ejercicio sirva como tutorial de introducción al análisis y diseño de sistemas de control robusto mediante el uso de la interfaz gráfica.

  20. PROMOCIÓN DE LA LECTURA E IDENTIDAD DEPORTIVA MEDIANTE TEXTOS DE HISTORIA DEL DEPORTE

    Directory of Open Access Journals (Sweden)

    José Guillermo Montero Quesada

    2016-04-01

    Full Text Available En el artículo se expone una experiencia de promoción de la lectura desarrollada mediante un proyecto conjunto de la Facultad de Cultura Física y la biblioteca de la Universidad de Las Tunas. Se fundamenta en aspectos teóricos de la promoción de la lectura y de la identidad deportiva, ambas variables tienen salida práctica en actividades donde confluyen diversas manifestaciones del arte y la literatura con la actividad física y el deporte. El objetivo del trabajo consiste en reflexionar en torno a la necesidad de asumir alternativas de cómo influir en la formación integral de los estudiantes universitarios de la carrera de Cultura Física, con énfasis en los conocimientos históricos y culturales.

  1. Predictor de Smith modificado mediante un modelo interno, robusto a perturbaciones externas no medibles.

    Directory of Open Access Journals (Sweden)

    Fernando Castillo García

    2010-11-01

    Full Text Available Normal 0 21 false false false MicrosoftInternetExplorer4 Se propone una modificación de la estructura del predictor de Smith mediante un modelo interno que posibilita aumentar su rechazo al efecto de las per-turbaciones externas no medibles en comparación con la estructura clásica del predictor de Smith. Los resultados obtenidos se aplican en el diseño de un sistema de control del proceso de variación de la temperatura del jugo en los calentadores de una fábrica de azúcar. Los resultados de la simulación del sistema diseñado mostraron su efectividad y robustez en cuanto al rechazo de perturbaciones externas no medibles. Palabras claves: Predictor de Smith, predictor de Smith modificado, rechazo a perturbaciones externas no medibles, robustez de los sistemas de control.

  2. Fomento de la conciencia ambiental mediante el blog UNAECOLÓGICA

    Directory of Open Access Journals (Sweden)

    Nereidy Velásquez

    2016-01-01

    Full Text Available El propósito del artículo es presentar una propuesta para fomentar la conciencia ambiental mediante el uso del blog. En este trabajo se organizaron los referentes teóricos relacionados con la educación ambiental y con la web, la cual constituye un espacio que ofrece información consistente y veraz sobre cualquier temática. El blog UNAECOLÓGICA es una propuesta que se crea con el fin de generar espacios para la comunicación, la interacción, la construcción del conocimiento ambiental y estimular en la comunidad unista la participación, la empatía y la solidaridad hacia su entorno.  Entre algunas de las secciones que presenta UNAECOLÓGICA, se encuentran: Notiambiente, Literambiente, Ecorelatos. Ecofrases, entre otras.

  3. DETECCIÓN Y AISLAMIENTO ROBUSTO DE FALLAS MEDIANTE OBSERVADORES CON ENTRADAS DESCONOCIDAS

    Directory of Open Access Journals (Sweden)

    JUAN ANZUREZ MARÍN

    2009-01-01

    Full Text Available En el presente artículo se muestra una metodología de diseño de observadores con entradas desconocidas para la solución del problema de Detección de Fallas. La técnica propuesta se basa principalmente en la observación de señales de error conocidas como residuos, las cuales se obtienen mediante la diferencia entre la salida actual del sistema y la salida estimada. Un observador con entradas desconocidas tiene la particularidad de que su vector de error de estimación tiende a cero asintóticamente, sin considerar la presencia de las entradas desconocidas o perturbaciones en el sistema. El algoritmo de detección se aplica satisfactoriamente en un sistema hidráulico de nivel de líquido tanto en simulación como en tiempo real.

  4. Generación de nuevos significados, mediante la metonimia, en el parlache

    Directory of Open Access Journals (Sweden)

    Juan Manuel Pérez Sánchez

    2009-04-01

    Full Text Available Reconociendo la incidencia del cambio semántico en el argot, se analiza aquí el papel de la metonimia como mecanismo de generación de nuevos significados en el argot juvenil de Medellín (Colombia, conocido como parlache. Como punto de partida, se delimita el concepto de argot y se clarifica lo que aquí se entiende por metonimia. Tras estas precisiones, se analiza una muestra de doce significados argóticos generados a partir de unidades ya existentes y mediante procesos metonímicos. Por último, como resultados, se observan tendencias dentro de la muestra en la utilización mayoritaria de ciertos tipos de metonimia. Así mismo, se vislumbra que en la muestra priman principios cognitivos universales como ''típico sobre atípico'', ''corporal sobre mental'', ''concreto sobre abstracto'', entre otros, a la hora de elegir los conceptos fuente.

  5. Comparison of kDNA PCR-hybridization assay with three PCR methods for canines visceral Leishmaniasis diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Pilatti, Marcia M.; Andrade, Antero S.R. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: marciapilatti@yahoo.com.br, e-mail: antero@cdtn.br; Ferreira, Sidney A. [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Dept. de Parasitologia], e-mail: saninoalmeida@gmail.com

    2009-07-01

    The sensitivity of the kDNA PCR-Hybridization assay, which uses radioactive DNA probes (labeled with {sup 32}P), was compared with three conventional PCR methods used for canine visceral leishmaniasis diagnosis. All PCR methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or semi nested). Two methods (kDNA PCR-Hybridization and kDNA snPCR) used primers addressed to kinetoplast minicircles and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of the ribosomal rRNA genes. The comparison was accomplished in two groups of 23 infected dogs using samples collected by the conjunctival swab procedure. In the Group 1 the DNA was extracted from cotton swabs by phenol-chloroform and in Group 2 by boiling. The most efficient PCR methods in the Group 1 were those based on kDNA targets. The kDNA PCR-Hybridization was able to detect parasites in 22/23 dogs (95.6%) and in 40/46 samples (86.9%). The kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The positivities of the kDNA based methods were significantly higher than the positivities verified for the methods based on ribosomal rRNA genes (p<0.05). In the Group 2 the kDNA PCR- Hybridization showed a better performance detecting parasites in 18/23 dogs (78.3%) and in 31/46 samples (67.4%), significantly higher than the other three methods (p<0.05). The higher sensitivity of the minicircle kDNA based assays reported by others was confirmed in this study and kDNA PCR-Hybridization showed the best sensitivity among the assays evaluated. (author)

  6. Comparison of kDNA PCR-hybridization assay with three PCR methods for canines visceral Leishmaniasis diagnosis

    International Nuclear Information System (INIS)

    Pilatti, Marcia M.; Andrade, Antero S.R.; Ferreira, Sidney A.

    2009-01-01

    The sensitivity of the kDNA PCR-Hybridization assay, which uses radioactive DNA probes (labeled with 32 P), was compared with three conventional PCR methods used for canine visceral leishmaniasis diagnosis. All PCR methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or semi nested). Two methods (kDNA PCR-Hybridization and kDNA snPCR) used primers addressed to kinetoplast minicircles and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of the ribosomal rRNA genes. The comparison was accomplished in two groups of 23 infected dogs using samples collected by the conjunctival swab procedure. In the Group 1 the DNA was extracted from cotton swabs by phenol-chloroform and in Group 2 by boiling. The most efficient PCR methods in the Group 1 were those based on kDNA targets. The kDNA PCR-Hybridization was able to detect parasites in 22/23 dogs (95.6%) and in 40/46 samples (86.9%). The kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The positivities of the kDNA based methods were significantly higher than the positivities verified for the methods based on ribosomal rRNA genes (p<0.05). In the Group 2 the kDNA PCR- Hybridization showed a better performance detecting parasites in 18/23 dogs (78.3%) and in 31/46 samples (67.4%), significantly higher than the other three methods (p<0.05). The higher sensitivity of the minicircle kDNA based assays reported by others was confirmed in this study and kDNA PCR-Hybridization showed the best sensitivity among the assays evaluated. (author)

  7. Medida de la dureza de sólidos mediante nanoindentación

    Directory of Open Access Journals (Sweden)

    Alkorta, J.

    2005-10-01

    Full Text Available Hardness is not readily measurable by means of instrumented indentation since the value of the contact area depends on the pile-up or sink-in occurring near the contact surface of the sample. The most widespread method to estimate it by means of the loading/unloading curve of indentation, Oliver and Pharr’s method, deviates, in the extreme cases, up to a 25% from the real values since it only takes into account the elastic deflection. In this work, a new correction based on Oliver and Pharr’s method is proposed that agrees with the numerical calculations. Plastic hardening behaviour of the sample must be known to accurately estimate the contact area.

    La medida de la dureza mediante indentación con registro de carga y desplazamiento no es evidente, dada la incertidumbre sobre el tamaño de huella debido al levantamiento (pile-up o hundimiento (sink-in plásticos de la superficie de la muestra alrededor del indentador. El método más utilizado para la medida de la dureza mediante la curva de carga/descarga de indentación, el de Oliver y Pharr, sólo tiene en cuenta hundimiento elástico, por lo que el error en la medida de la dureza y el módulo de Young puede llegar hasta un 25% en los casos más extremos. En el presente trabajo se discute una posible corrección al método de Oliver y Pharr para una obtención más ajustada del área de contacto de la huella. Esta corrección requiere de un conocimiento a priori o a posteriori del comportamiento plástico del material.

  8. Study On Application Of Molecular Techniques (RAPD-PCR And RAMP-PCR) To Detect Mutation In Rice Breeding

    International Nuclear Information System (INIS)

    Hoang Thi My Linh; Phan, D. T. Son; Nguyen Thi Vang; Nguyen, T. T. Hien; Le XuanTham

    2007-01-01

    The project was carried out in 2007 with the purpose of consideration for using the two simple and inexpensive molecular techniques to estimate changes in DNA of rice mutant after gamma irradiation. Three rice cultivars: Basmati370, Tam Thom (TT1), IR64 and three gamma irradiated mutants BDS, TDS and VND 95-20 respectively, were used. Suitable DNA extraction procedure was obtained. PCR optimization was conducted on three important factors including: amount of MgCl 2 , DNA concentration and annealing temperature. 2.5 mM of MgCl 2 for RAPD-PCR and 3.75 mM for RAMP-PCR were found the best. 40 ng DNA provided a good amplification for RAMP-PCR; this figure was 50 ng for RAPD-PCR. Annealing temperatures were determined at 36 o C for RAPD primer and at 55±3 o C for Microsatellite primer. Final results showed that, both RAPD-PCR and RAMP-PCR could detect changes in DNA of rice mutants after gamma irradiation compared to their parents. Percentage of DNA changes determined by RAPD-PCR and RAMP-PCR on Basmati370 and its mutant BDS were 11.49% and 21.2% respectively; These on TT1 and TDS were 8.98% and 15.4%; and on IR64 and VND 95-20 were 3.45% and 4.95%. (author)

  9. Comparison of ELISA, nested PCR and sequencing and a novel qPCR for detection of Giardia isolates from Jordan.

    Science.gov (United States)

    Hijjawi, Nawal; Yang, Rongchang; Hatmal, Ma'mon; Yassin, Yasmeen; Mharib, Taghrid; Mukbel, Rami; Mahmoud, Sameer Alhaj; Al-Shudifat, Abdel-Ellah; Ryan, Una

    2018-02-01

    Little is known about the prevalence of Giardia duodenalis in human patients in Jordan and all previous studies have used direct microscopy, which lacks sensitivity. The present study developed a novel quantitative PCR (qPCR) assay at the β-giardin (bg) locus and evaluated its use as a frontline test for the diagnosis of giardiasis in comparison with a commercially available ELISA using nested PCR and sequencing of the glutamate dehydrogenase (gdh) locus (gdh nPCR) as the gold standard. A total of 96 human faecal samples were collected from 96 patients suffering from diarrhoea from 5 regions of Jordan and were screened using the ELISA and qPCR. The analytical specificity of the bg qPCR assay revealed no cross-reactions with other genera and detected all the Giardia isolates tested. Analytical sensitivity was 1 Giardia cyst per μl of DNA extract. The overall prevalence of Giardia was 64.6%. The clinical sensitivity and specificity of the bg qPCR was 89.9% and 82.9% respectively compared to 76.5 and 68.0% for the ELISA. This study is the first to compare three different methods (ELISA, bg qPCR, nested PCR and sequencing at the gdh locus) to diagnose Jordanian patients suffering from giardiasis and to analyze their demographic data. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Procedimiento para la obtención de levaduras vínicas superproductoras de manoproteínas mediante tecnologías no recombinantes

    OpenAIRE

    Barcenilla Moraleda, José María; González Ramos, Daniel; Tabera, Laura; González García, Ramón

    2008-01-01

    Procedimiento para la obtención de levaduras vínicas superproductoras de manoproteínas mediante tecnologías no recombinantes. Procedimiento para obtener cepas de levaduras superproductoras de manoproteínas mediante la selección de mutantes resistentes a la toxina K9, cepas obtenibles por dicho procedimiento y usos.

  11. Superficie específica de una bentonita mediante la adsorción de azul de metileno

    OpenAIRE

    Pinzón Bello, Jorge Alejo

    2010-01-01

    Se estudió la determinación de la superficie específica de una bentonita colombiana, procedente del Valle del Cauca, mediante la adsorción de azul de metileno, a 298 K. Este método se comparó con el de la adsorción de nitrógeno a 77 K.

  12. Human fecal source identification with real-time quantitative PCR

    Science.gov (United States)

    Waterborne diseases represent a significant public health risk worldwide, and can originate from contact with water contaminated with human fecal material. We describe a real-time quantitative PCR (qPCR) method that targets a Bacteroides dori human-associated genetic marker for...

  13. Comparison between ICT and PCR for diagnosis of Chlamydia trachomatis.

    Science.gov (United States)

    Khan, E R; Hossain, M A; Paul, S K; Mahmud, C; Hasan, M M; Rahman, M M; Nahar, K; Kubayashi, N

    2012-04-01

    Chlamydia trachomatis is an obligate intracellular gram-negative bacterium which is the most prevalent cause of bacterial sexually transmitted infections (STI). The present study was carried to diagnose genital Chlamydia trachomatis infection among women of reproductive age, attending Mymensingh Medical College Hospital, during July 2009 to June 2010 by Immunochromatographic test (ICT) and Polymerase chain reaction (PCR). A total of 70 females were included in this study. Out of 70 cases 56 were symptomatic and 14 asymptomatic. Endocervical swabs were collected from each of the cases and examined by Immunochromatographic test (ICT) for antigen detection and Polymerase chain reaction (PCR) for detection of endogenous plasmid-based nucleic acid. A total 29(41.4%) of the cases were found positive for C. trachomatis either by ICT or PCR. Of the 56 symptomatic cases, 19(33.9%) were found ICT positive and 17(30.4%) were PCR positive. Among 14 asymptomatic females, 2(14.3%) were ICT positive and none were PCR positive. Though PCR is highly sensitive but a total of twelve cases were found ICT positive but PCR negative. It may be due to presence of plasmid deficient strain of C trachomatis which could be amplified by ompA based (Chromosomal gene) multiplex PCR.

  14. Comparison of Nested-PCR technique and culture method in ...

    African Journals Online (AJOL)

    USER

    2010-04-05

    Apr 5, 2010 ... Full Length Research Paper. Comparison of ... The aim of the present study was to evaluate the diagnostic value of nested PCR in genitourinary ... method. Based on obtained results, the positivity rate of urine samples in this study was 5.0% by using culture and PCR methods and 2.5% for acid fast staining.

  15. Development of a competitive PCR assay for the quantification of ...

    African Journals Online (AJOL)

    The E. coli malate dehydrogenase Mdh house-keeping gene was modified and used as an internal control and competitor DNA for the c-PCR. E. coli cell concentration equivalents ranging from 20 to 2 x 104 cells ml-1 could be quantified with the c-PCR. Fifty-three water samples from various sources were tested with the ...

  16. Evaluation of PCR for diagnosis of visceral leishmaniasis

    NARCIS (Netherlands)

    Osman, O. F.; Oskam, L.; Zijlstra, E. E.; Kroon, N. C.; Schoone, G. J.; Khalil, E. T.; El-Hassan, A. M.; Kager, P. A.

    1997-01-01

    An evaluation of Leishmania PCR was performed with bone marrow, lymph node, and blood samples from 492 patients, 60 positive controls, and 90 negative controls. Results were compared with microscopy results for Giemsa-stained smears. PCR and microscopy of lymph node and bone marrow aspirates from

  17. Digital PCR for direct quantification of viruses without DNA extraction.

    Science.gov (United States)

    Pavšič, Jernej; Žel, Jana; Milavec, Mojca

    2016-01-01

    DNA extraction before amplification is considered an essential step for quantification of viral DNA using real-time PCR (qPCR). However, this can directly affect the final measurements due to variable DNA yields and removal of inhibitors, which leads to increased inter-laboratory variability of qPCR measurements and reduced agreement on viral loads. Digital PCR (dPCR) might be an advantageous methodology for the measurement of virus concentrations, as it does not depend on any calibration material and it has higher tolerance to inhibitors. DNA quantification without an extraction step (i.e. direct quantification) was performed here using dPCR and two different human cytomegalovirus whole-virus materials. Two dPCR platforms were used for this direct quantification of the viral DNA, and these were compared with quantification of the extracted viral DNA in terms of yield and variability. Direct quantification of both whole-virus materials present in simple matrices like cell lysate or Tris-HCl buffer provided repeatable measurements of virus concentrations that were probably in closer agreement with the actual viral load than when estimated through quantification of the extracted DNA. Direct dPCR quantification of other viruses, reference materials and clinically relevant matrices is now needed to show the full versatility of this very promising and cost-efficient development in virus quantification.

  18. Inhibitory effect of common microfluidic materials on PCR outcome

    KAUST Repository

    Kodzius, Rimantas

    2013-10-10

    In this study, we established a simple method for evaluating the PCR compatibility of various common materials employed when fabricating microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most cases, adding bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, although they noticeably interacted with the polymerase. We provide a simple method of performing PCR-compatibility testing of materials using inexpensive instrumentation that is common in molecular biology laboratories. Furthermore, our method is direct, being performed under actual PCR conditions with high temperature. Our results provide an overview of materials that are PCR-friendly for fabricating microfluidic devices. The PCR reaction, without any additives, performed best with pyrex glass, and it performed worst with PMMA or acrylic glue materials.

  19. Detection of Bacillus spores using PCR and FTA filters.

    Science.gov (United States)

    Lampel, Keith A; Dyer, Deanne; Kornegay, Leroy; Orlandi, Palmer A

    2004-05-01

    Emphasis has been placed on developing and implementing rapid detection systems for microbial pathogens. We have explored the utility of expanding FTA filter technology for the preparation of template DNA for PCR from bacterial spores. Isolated spores from several Bacillus spp., B. subtilis, B. cereus, and B. megaterium, were applied to FTA filters, and specific DNA products were amplified by PCR. Spore preparations were examined microscopically to ensure that the presence of vegetative cells, if any, did not yield misleading results. PCR primers SRM86 and SRM87 targeted a conserved region of bacterial rRNA genes, whereas primers Bsub5F and Bsub3R amplified a product from a conserved sequence of the B. subtilis rRNA gene. With the use of the latter set of primers for nested PCR, the sensitivity of the PCR-based assay was increased. Overall, 53 spores could be detected after the first round of PCR, and the sensitivity was increased to five spores by nested PCR. FTA filters are an excellent platform to remove PCR inhibitors and have universal applications for environmental, clinical, and food samples.

  20. Inhibitory effect of common microfluidic materials on PCR outcome

    KAUST Repository

    Kodzius, Rimantas; Xiao, Kang; Wu, Jinbo; Yi, Xin; Gong, Xiuqing; Foulds, Ian G.; Wen, Weijia

    2013-01-01

    In this study, we established a simple method for evaluating the PCR compatibility of various common materials employed when fabricating microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most cases, adding bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, although they noticeably interacted with the polymerase. We provide a simple method of performing PCR-compatibility testing of materials using inexpensive instrumentation that is common in molecular biology laboratories. Furthermore, our method is direct, being performed under actual PCR conditions with high temperature. Our results provide an overview of materials that are PCR-friendly for fabricating microfluidic devices. The PCR reaction, without any additives, performed best with pyrex glass, and it performed worst with PMMA or acrylic glue materials.

  1. Splinkerette PCR for mapping transposable elements in Drosophila.

    Directory of Open Access Journals (Sweden)

    Christopher J Potter

    2010-04-01

    Full Text Available Transposable elements (such as the P-element and piggyBac have been used to introduce thousands of transgenic constructs into the Drosophila genome. These transgenic constructs serve many roles, from assaying gene/cell function, to controlling chromosome arm rearrangement. Knowing the precise genomic insertion site for the transposable element is often desired. This enables identification of genomic enhancer regions trapped by an enhancer trap, identification of the gene mutated by a transposon insertion, or simplifying recombination experiments. The most commonly used transgene mapping method is inverse PCR (iPCR. Although usually effective, limitations with iPCR hinder its ability to isolate flanking genomic DNA in complex genomic loci, such as those that contain natural transposons. Here we report the adaptation of the splinkerette PCR (spPCR method for the isolation of flanking genomic DNA of any P-element or piggyBac. We report a simple and detailed protocol for spPCR. We use spPCR to 1 map a GAL4 enhancer trap located inside a natural transposon, pinpointing a master regulatory region for olfactory neuron expression in the brain; and 2 map all commonly used centromeric FRT insertion sites. The ease, efficiency, and efficacy of spPCR could make it a favored choice for the mapping of transposable element in Drosophila.

  2. Multiplex PCR identification of Taenia spp. in rodents and carnivores.

    Science.gov (United States)

    Al-Sabi, Mohammad N S; Kapel, Christian M O

    2011-11-01

    The genus Taenia includes several species of veterinary and public health importance, but diagnosis of the etiological agent in definitive and intermediate hosts often relies on labor intensive and few specific morphometric criteria, especially in immature worms and underdeveloped metacestodes. In the present study, a multiplex PCR, based on five primers targeting the 18S rDNA and ITS2 sequences, produced a species-specific banding patterns for a range of Taenia spp. Species typing by the multiplex PCR was compared to morphological identification and sequencing of cox1 and/or 12S rDNA genes. As compared to sequencing, the multiplex PCR identified 31 of 32 Taenia metacestodes from rodents, whereas only 14 cysts were specifically identified by morphology. Likewise, the multiplex PCR identified 108 of 130 adult worms, while only 57 were identified to species by morphology. The tested multiplex PCR system may potentially be used for studies of Taenia spp. transmitted between rodents and carnivores.

  3. [A new method of processing quantitative PCR data].

    Science.gov (United States)

    Ke, Bing-Shen; Li, Guang-Yun; Chen, Shi-Min; Huang, Xiang-Yan; Chen, Ying-Jian; Xu, Jun

    2003-05-01

    Today standard PCR can't satisfy the need of biotechnique development and clinical research any more. After numerous dynamic research, PE company found there is a linear relation between initial template number and cycling time when the accumulating fluorescent product is detectable.Therefore,they developed a quantitative PCR technique to be used in PE7700 and PE5700. But the error of this technique is too great to satisfy the need of biotechnique development and clinical research. A better quantitative PCR technique is needed. The mathematical model submitted here is combined with the achievement of relative science,and based on the PCR principle and careful analysis of molecular relationship of main members in PCR reaction system. This model describes the function relation between product quantity or fluorescence intensity and initial template number and other reaction conditions, and can reflect the accumulating rule of PCR product molecule accurately. Accurate quantitative PCR analysis can be made use this function relation. Accumulated PCR product quantity can be obtained from initial template number. Using this model to do quantitative PCR analysis,result error is only related to the accuracy of fluorescence intensity or the instrument used. For an example, when the fluorescence intensity is accurate to 6 digits and the template size is between 100 to 1,000,000, the quantitative result accuracy will be more than 99%. The difference of result error is distinct using same condition,same instrument but different analysis method. Moreover,if the PCR quantitative analysis system is used to process data, it will get result 80 times of accuracy than using CT method.

  4. Comparison of droplet digital PCR and seminested real-time PCR for quantification of cell-associated HIV-1 RNA

    NARCIS (Netherlands)

    Kiselinova, Maja; Pasternak, Alexander O.; de Spiegelaere, Ward; Vogelaers, Dirk; Berkhout, Ben; Vandekerckhove, Linos

    2014-01-01

    Cell-associated (CA) HIV-1 RNA is considered a potential marker for assessment of viral reservoir dynamics and antiretroviral therapy (ART) response in HIV-infected patients. Recent studies employed sensitive seminested real-time quantitative (q)PCR to quantify CA HIV-1 RNA. Digital PCR has been

  5. Transient neonatal diabetes mellitus with macroglossia diagnosed by methylation specific PCR (MS-PCR

    Directory of Open Access Journals (Sweden)

    Hye Young Jin

    2010-03-01

    Full Text Available Transient neonatal diabetes mellitus (TNDM has been associated with paternal uniparental isodisomy of chromosome 6, paternally inherited duplication of 6q24, or a methylation defect at a CpG island of the ZAC or HYMAI gene. We experienced a case of TNDM in which the patient presented with hyperglycemia, macroglossia, and intrauterine growth retardation, caused by a paternally derived HYMAI. An 18-day-old female infant was admitted to the hospital because of macroglossia and recurrent hyperglycemia. In addition to the macroglossia, she also presented with large fontanelles, micrognathia, and prominent eyes. Serum glucose levels were 200&#8211;300 mg/dL and they improved spontaneously 2 days after admission. To identify the presence of a maternal methylated allele, bisulfite-treated genomic DNA from peripheral blood was prepared and digested with BssHII after polymerase chain reaction (PCR amplification with methylation-specific HYMAI primers. PCR and restriction fragment length polymorphism analysis showed that the patient had only the paternal origin of the HYMA1 gene. TNDM is associated with a methylation defect in chromosome 6, suggesting that an imprinted gene on chromosome 6 is responsible for this phenotype.

  6. Recubrimientos de TiN depositados mediante ACPVD sobre aleaciones de magnesio AM60

    Directory of Open Access Journals (Sweden)

    Pichel, M.

    2013-06-01

    Full Text Available Magnesium alloys are reaching special interest due to their good specific properties, low cost and good manufacturing properties. However, their low hardness, wear and corrosion resistance limit their applications in certain sectors of industry. These drawbacks can be solved by applying hard ceramic coatings, such as nitrides or metal carbides. TiN is one of the most used coatings due to its high adhesion, hardness, low coefficient of friction and chemical stability. Physical vapor deposition by cathodic arc CAPVD, is a versatile technique, which uses low temperatures and high ionization energies, generating homogeneous coatings. To achieve coatings with high quality, a careful control of the manufacturing parameters is required, such as bias voltage, gas flow or intensity. This paper focuses on magnesium alloys, AM60, coated with TiN using physical vapor deposition cathodic arc technique (CAPVD at different intensity values (40A and 100A and surface preparation (grinding up to 4000 grit and polished to 3μm. It was included a final condition with an intermediate Al film. The samples were characterized by X-ray diffraction, roughness, optical microscopy and scanning electron.Las aleaciones de magnesio están alcanzando especial interés gracias a sus buenas propiedades específicas, bajo coste y buenas propiedades de moldeabilidad. No obstante su baja dureza, resistencia a desgaste y corrosión, limita sus aplicaciones en ciertos campos de la industria. Estos inconvenientes se pueden solucionar aplicando recubrimientos duros cerámicos, como nitruros o carburos metálicos. El TiN es uno de los más utilizados debido a su alta adherencia, dureza, bajo coeficiente de fricción y estabilidad química. La deposición física en fase vapor mediante arco catódico, ACPVD, es una técnica muy versátil, que emplea bajas temperaturas y altas energías de ionización, generando recubrimientos de bajo espesor, homogéneos y de calidad. Para alcanzar

  7. RT-PCR Protocols - Methods in Molecular Biology

    Directory of Open Access Journals (Sweden)

    Manuela Monti

    2011-03-01

    Full Text Available “The first record I have of it, is when I made a computer file which I usually did whenever I had an idea, that would have been on the Monday when I got back, and I called it Chain Reaction.POL, meaning polymerase. That was the identifier for it and later I called the thing the Polymerase Chain Reaction, which a lot of people thought was a dumb name for it, but it stuck, and it became PCR”. With these words the Nobel prize winner, Kary Mullis, explains how he named the PCR: one of the most important techniques ever invented and currently used in molecular biology. This book “RT-PCR Protocols” covers a wide range of aspects important for the setting of a PCR experiment for both beginners and advanced users. In my opinion the book is very well structured in three different sections. The first one describes the different technologies now available, like competitive RT-PCR, nested RT-PCR or RT-PCR for cloning. An important part regards the usage of PCR in single cell mouse embryos, stressing how important...........

  8. Shape based kinetic outlier detection in real-time PCR

    Directory of Open Access Journals (Sweden)

    D'Atri Mario

    2010-04-01

    Full Text Available Abstract Background Real-time PCR has recently become the technique of choice for absolute and relative nucleic acid quantification. The gold standard quantification method in real-time PCR assumes that the compared samples have similar PCR efficiency. However, many factors present in biological samples affect PCR kinetic, confounding quantification analysis. In this work we propose a new strategy to detect outlier samples, called SOD. Results Richards function was fitted on fluorescence readings to parameterize the amplification curves. There was not a significant correlation between calculated amplification parameters (plateau, slope and y-coordinate of the inflection point and the Log of input DNA demonstrating that this approach can be used to achieve a "fingerprint" for each amplification curve. To identify the outlier runs, the calculated parameters of each unknown sample were compared to those of the standard samples. When a significant underestimation of starting DNA molecules was found, due to the presence of biological inhibitors such as tannic acid, IgG or quercitin, SOD efficiently marked these amplification profiles as outliers. SOD was subsequently compared with KOD, the current approach based on PCR efficiency estimation. The data obtained showed that SOD was more sensitive than KOD, whereas SOD and KOD were equally specific. Conclusion Our results demonstrated, for the first time, that outlier detection can be based on amplification shape instead of PCR efficiency. SOD represents an improvement in real-time PCR analysis because it decreases the variance of data thus increasing the reliability of quantification.

  9. IDENTIFIKASI TIPE HLA KELAS II DENGAN TEKNIK PCR

    Directory of Open Access Journals (Sweden)

    Ervi Salwati

    2012-09-01

    Full Text Available HLA (Human Leukocyte Antigen contains a set of genes located together on the short arm of chromosome 6. These genes control immune responses, graft acceptance or rejection and tumor surveillance. These abilities have close relationship with genetic variation (occur in "many forms" or alleles that bind and present antigens to T lymphocytes. Using advanced technology and molecular biology approaches (PCR technique detection of genetic variation in the HLA region (or HLA typing has been performed based on DNA.. PCR is an in vitro technique to amplify the DNA sequence enzymatically. "Sequence Specific Primers" (SSP are designed for this PCR to obtain amplification of specific alleles or groups of alleles. The PCR products are visualized through agarose gel electrophoresis stained with ethidium bromide. The PCR technique requires small amount of whole blood (0.5 - 1 ml, gives rapid, accurate and complete result. This paper discuss identification of HLA class II typing using PCR-SSP technique and show the examples of the results.   Key words: HLA (Human Leukocyte Antigen class II, PCR (Polymerase Chain Reaction

  10. Broad-range PCR: past, present, or future of bacteriology?

    Science.gov (United States)

    Renvoisé, A; Brossier, F; Sougakoff, W; Jarlier, V; Aubry, A

    2013-08-01

    PCR targeting the gene encoding 16S ribosomal RNA (commonly named broad-range PCR or 16S PCR) has been used for 20 years as a polyvalent tool to study prokaryotes. Broad-range PCR was first used as a taxonomic tool, then in clinical microbiology. We will describe the use of broad-range PCR in clinical microbiology. The first application was identification of bacterial strains obtained by culture but whose phenotypic or proteomic identification remained difficult or impossible. This changed bacterial taxonomy and allowed discovering many new species. The second application of broad-range PCR in clinical microbiology is the detection of bacterial DNA from clinical samples; we will review the clinical settings in which the technique proved useful (such as endocarditis) and those in which it did not (such as characterization of bacteria in ascites, in cirrhotic patients). This technique allowed identifying the etiological agents for several diseases, such as Whipple disease. This review is a synthesis of data concerning the applications, assets, and drawbacks of broad-range PCR in clinical microbiology. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  11. Ureaplasma parvum prosthetic joint infection detected by PCR.

    Science.gov (United States)

    Farrell, John J; Larson, Joshua A; Akeson, Jeffrey W; Lowery, Kristin S; Rounds, Megan A; Sampath, Rangarajan; Bonomo, Robert A; Patel, Robin

    2014-06-01

    We describe the first reported case of Ureaplasma parvum prosthetic joint infection (PJI) detected by PCR. Ureaplasma species do not possess a cell wall and are usually associated with colonization and infection of mucosal surfaces (not prosthetic material). U. parvum is a relatively new species name for certain serovars of Ureaplasma urealyticum, and PCR is useful for species determination. Our patient presented with late infection of his right total knee arthroplasty. Intraoperative fluid and tissue cultures and pre- and postoperative synovial fluid cultures were all negative. To discern the pathogen, we employed PCR coupled with electrospray ionization mass spectrometry (PCR/ESI-MS). Our patient's failure to respond to empirical antimicrobial treatment and our previous experience with PCR/ESI-MS in culture-negative cases of infection prompted us to use this approach over other diagnostic modalities. PCR/ESI-MS detected U. parvum in all samples. U. parvum-specific PCR testing was performed on all synovial fluid samples to confirm the U. parvum detection. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  12. DNA Differential Diagnosis of Taeniasis and Cysticercosis by Multiplex PCR

    Science.gov (United States)

    Yamasaki, Hiroshi; Allan, James C.; Sato, Marcello Otake; Nakao, Minoru; Sako, Yasuhito; Nakaya, Kazuhiro; Qiu, Dongchuan; Mamuti, Wulamu; Craig, Philip S.; Ito, Akira

    2004-01-01

    Multiplex PCR was established for differential diagnosis of taeniasis and cysticercosis, including their causative agents. For identification of the parasites, multiplex PCR with cytochrome c oxidase subunit 1 gene yielded evident differential products unique for Taenia saginata and Taenia asiatica and for American/African and Asian genotypes of Taenia solium with molecular sizes of 827, 269, 720, and 984 bp, respectively. In the PCR-based detection of tapeworm carriers using fecal samples, the diagnostic markers were detected from 7 of 14 and 4 of 9 T. solium carriers from Guatemala and Indonesia, respectively. Test sensitivity may have been reduced by the length of time (up to 12 years) that samples were stored and/or small sample volumes (ca. 30 to 50 mg). However, the diagnostic markers were detected by nested PCR in five worm carriers from Guatemalan cases that were found to be negative by multiplex PCR. It was noteworthy that a 720 bp-diagnostic marker was detected from a T. solium carrier who was egg-free, implying that it is possible to detect worm carriers and treat before mature gravid proglottids are discharged. In contrast to T. solium carriers, 827-bp markers were detected by multiplex PCR in all T. saginata carriers. The application of the multiplex PCR would be useful not only for surveillance of taeniasis and cysticercosis control but also for the molecular epidemiological survey of these cestode infections. PMID:14766815

  13. Tratamiento del polvo de aluminio mediante disolución acuosa

    Directory of Open Access Journals (Sweden)

    López, F. A.

    2004-10-01

    Full Text Available Aluminium dust from aluminium remelting industry is a hazardous residue because of its high reactivity in the presence of water. In order to apply the new European Directive about landfill of waste, a study of its hydrolysis was carried out. The influence of temperature, time and pH on the hydrolysis of the aluminium dust was studied. The hydrolysed solids were characterized by XRD and AAS; in the aqueous solutions the pH and the ionic conductivity were determined. The evolved gases were analysed by mass spectrometry. The reactivity of the dust, before and after hydrolysis, was investigated by analysing the ammonia, hydrogen sulphide and metallic aluminium. By hydrolysis at 60 °C and 48 h a much lower reactive material was obtained which could be disposed with minimal environmental impact.

    El polvo de aluminio es un residuo generado en la metalurgia secundaria del aluminio y considerado peligroso como consecuencia de su elevada reactividad en presencia de humedad. Con objetivo de buscar un procedimiento de pretratamiento de dicho residuo, de acuerdo con la Directiva Europea sobre vertederos, se ha realizado el estudio del comportamiento del polvo de aluminio en medio acuoso. Para ello, se han analizado la influencia de la temperatura, el tiempo y el pH de reacción en su hidrólisis. Los sólidos hidrolizados se caracterizaron mediante EAA y DRX, mientras que en las soluciones acuosas resultantes se determinaron el pH y la conductividad iónica. Los gases liberados durante el proceso de hidrólisis se analizaron mediante espectrometría de masas. Asimismo, se ha determinado la reactividad del residuo antes y después de la hidrólisis, analizando amoniaco, sulfuro de hidrógeno y aluminio metálico. La hidrólisis, a 60 °C y después de 48 h, permite obtener material de muy baja reactividad que podría ser almacenado en vertedero.

  14. Estudio mediante afm de estructuras de silicalita para la separación de gases

    Directory of Open Access Journals (Sweden)

    Prádanos, P.

    2004-02-01

    Full Text Available In this work, we study films of silicalite crystals used in gas separation processes. These crystals were obtained by hydrothermal synthesis during different times and using different gel composition. They were deposited on an alumina support growing in two preferential directions. Finally, the material was placed in a stove at 480ºC during 8 h in order to remove the structurant agent with heating and cooling rates of 0.5 y 2 ºC/min respectively. The resulting surfaces were analysed by atomic force microscopy (AFM in tapping mode in order to notice the deposition of the silicalite crystals in the indicated directions, and also to distinguish the evolution of the nuclei growth. At the same time, the porous structure of silicalite has been determined, leading to results in good agreement with those obtained by other techniques.

    En este trabajo se han estudiado láminas de silicalita con aplicación en los procesos de separación de gases. Dichas láminas se han depositado mediante síntesis hidrotermal durante distintos tiempos y usando varias composiciones en el gel precursor. La deposición se hizo sobre un soporte de alúmina con crecimiento preferencial en dos direcciones. Finalmente las láminas se calcinaron a 753 K durante 8 h para eliminar el agente estructurante, usando velocidades de calentamiento y enfriamiento de 0.5 y 1 K/min respectivamente. Las superficies resultantes se han analizado mediante microscopía de fuerza atómica en modo de contacto-intermitente (tapping con el fin de ver si efectivamente se ha conseguido depositar cristales de silicalita en las direcciones indicadas y seguir la evolución de crecimiento de los núcleos. Por otro lado, se ha determinado la estructura porosa de la silicalita comprobando que los resultados concuerdan con los obtenidos por otras técnicas.

  15. Microestructura y propiedades de capas de tribaloy T-800 depositadas mediante plaqueado láser

    Directory of Open Access Journals (Sweden)

    Navas, C.

    2004-04-01

    Full Text Available The present work is based on obtaining Co based coatings (Tribaloy T-800 on plates of 18/8 stainless steel (AISI 304 by laser cladding technique. After the treatment, samples were characterized by optical and scanning electron microscopy with EDS analysis. The elemental composition of the coating was determined with a glow discharge lamp spectrometer (GDL. The study of the interface revealed a good adherence between the substrate and coating without substantial defects. For the laser cladded coatings, the microstructure was dendritic with a high degree of refinement and chemical homogeneity close to the original powder. The grain coarsening was observed in the overlapping zones due to the second heat treatment. Microhardness of the coatings reached 750 HV, a considerably higher value than the substrate hardness (200 HV. Also, the coating corrosion resistance in saline solutions had a great improvement.

    El presente trabajo se centra en la obtención de capas de aleación base Cobalto (Tribaloy T-800 sobre un sustrato de acero inoxidable 18/8 (AISI 304 mediante la técnica de plaqueado láser. Tras los tratamientos, se caracterizaron las probetas mediante microscopía óptica y electrónica de barrido con microanálisis (EDS. La composición elemental del recubrimiento se determinó en un espectrómetro de emisión óptica con fuente de excitación (GDL. El estudio de la interfase reveló una perfecta adherencia entre el recubrimiento y el material base, sin defectos apreciables. La microestructura de las capas depositadas es dendrítica con un alto grado de refinamiento y una homogeneidad química a lo largo del cordón y con valores muy próximos a los del polvo de partida. En las zonas de solape entre cordones, se observó un crecimiento del grano debido al segundo tratamiento térmico recibido. La microdureza de las capas alcanza los 750 HV, valor considerablemente superior al del sustrato (200 HV. Asimismo, se obtuvo una mejora

  16. PCR-RFLP y RAPD para la tipificación de Leishmania neotropical

    Directory of Open Access Journals (Sweden)

    Ana Margarita Montalvo

    2008-12-01

    Full Text Available Introducción. El análisis de la longitud de los fragmentos de restricción del producto amplificado y el estudio del ADN polimórfico amplificado al azar han demostrado ser herramientas útiles para la tipificación de Leishmania. Objetivos. Estudiar la utilidad de las técnicas moleculares para la identificación y tipificación de cepas de referencia de Leishmania spp. del Nuevo Mundo y valorar su aplicabilidad a muestras clínicas. Materiales y métodos. Se aplicó PCR para amplificar el gen que codifica la cisteíno-proteinasa B, y el análisis de la longitud de los fragmentos de restricción del producto amplificado utilizando ácido desoxirribonucleico de 16 cepas de referencia de Latinoamérica y de muestras clínicas de pacientes colombianos con leishmaniasis, y la técnica del ácido desoxirribonucleico polimórfico amplificado al azar utilizando ocho cepas de referencia. Se establecieron los patrones de bandas en cada caso. Resultados. Se obtuvo producto de amplificación en la PCR para Leishmania braziliensis, L. peruviana, L. panamensis y L. guyanensis. Para el resto, no fue posible amplificar el gen con los cebadores utilizados. La restricción mostró un patrón de bandas común para L. peruviana, L. guyanensis y L. panamensis, mientras L. braziliensis, presentaba un perfil individual único. El análisis de restricción del producto amplificado generó un patrón de bandas similar en los cinco pacientes estudiados, que se correspondía con el patrón generado por L. peruviana, L. guyanensis o L. panamensis. Mediante la amplificación al azar se obtuvieron patrones de bandas reproducibles con todas las cepas estudiadas, que posibilitaron la diferenciación. Se discuten las ventajas y limitaciones de ambos procederes. Conclusiones. El combinar ambas metodologías resultaría útil para identificar especies de importancia médica, tomando en cuenta sus ventajas y desventajas.

  17. A multiplex PCR for detection of six viruses in ducks.

    Science.gov (United States)

    Wang, Yongjuan; Zhu, Shanyuan; Hong, Weiming; Wang, Anping; Zuo, Weiyong

    2017-10-01

    In this study, six pairs of specific primers that can amplify DNA fragments of different sizes were designed and synthesized according to viral protein gene sequences published in GenBank. Then, a multiplex PCR method was established for rapid detection of duck hepatitis virus 1, duck plague virus, duck Tembusu virus, muscovy duck parvovirus, muscovy duck reovirus, and duck H9N2 avian influenza virus, and achieve simple and rapid detection of viral diseases in ducks. Single PCR was used to confirm primer specificity, and PCR conditions were optimized to construct a multiplex PCR system. Specificity and sensitivity assays were also developed. The multiplex PCR was used to detect duck embryos infected with mixed viruses and those with clinically suspected diseases to verify the feasibility of the multiplex PCR. Results show that the primers can specifically amplify target fragments, without any cross-amplification with other viruses. The multiplex PCR system can amplify six DNA fragments from the pooled viral genomes and specifically detect nucleic acids of the six duck susceptible viruses when the template amount is 10 2 copies/μl. In addition, the system can be used to detect viral nucleic acids in duck embryos infected with the six common viruses. The detection results for clinical samples are consistent with those detected by single PCR. Therefore, the established multiplex PCR method can perform specific, sensitive, and high-throughput detection of six duck-infecting viruses and can be applied to clinical identification and diagnosis of viral infection in ducks. Copyright © 2017. Published by Elsevier B.V.

  18. PCR IN TRAUMATOLOGY AND ORTHOPAEDICS: METHOD DESCRIPTION AND APPLICABILITY

    Directory of Open Access Journals (Sweden)

    E. M. Polyakova

    2014-01-01

    Full Text Available Review brief presents description of polymerase chain reaction method (PCR and its most common variants. Three PCR-based lines of research, carried out in the traumatology and orthopaedics, include identifying a causative agents of the implant-associated infection after orthopaedic surgery; detection of antibiotic resistance genes and biofilm forming genes. It was shown that PCR can be used as additional method for detection of genetic disorders, significant for traumatology and orthopaedics, and for investigation of cartilage and bone regeneration.

  19. PCR amplification on microarrays of gel immobilized oligonucleotides

    Science.gov (United States)

    Strizhkov, Boris; Tillib, Sergei; Mikhailovich, Vladimir; Mirzabekov, Andrei

    2003-11-04

    The invention relates two general methods for performing PCR amplification, combined with the detection and analysis of the PCR products on a microchip. In the first method, the amplification occurs both outside and within a plurality of gel pads on a microchip, with at least one oligonucleotide primer immobilized in a gel pad. In the second method, PCR amplification also takes place within gel pads on a microchip, but the pads are surrounded by a hydrophobic liquid such as that which separates the individual gel pads into environments which resemble micro-miniaturized test tubes.

  20. Sensitive simultaneous detection of seven sexually transmitted agents in semen by multiplex-PCR and of HPV by single PCR.

    Directory of Open Access Journals (Sweden)

    Fabrícia Gimenes

    Full Text Available Sexually transmitted diseases (STDs may impair sperm parameters and functions thereby promoting male infertility. To date limited molecular studies were conducted to evaluate the frequency and type of such infections in semen Thus, we aimed at conceiving and validating a multiplex PCR (M-PCR assay for the simultaneous detection of the following STD pathogens in semen: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Herpes virus simplex (HSV -1 and -2, and Treponema pallidum; We also investigated the potential usefulness of this M-PCR assay in screening programs for semen pathogens. In addition, we aimed: to detect human Papillomavirus (HPV and genotypes by single PCR (sPCR in the same semen samples; to determine the prevalence of the seven STDs, HPV and co-infections; to assess the possibility that these infections affect semen parameters and thus fertility. The overall validation parameters of M-PCR were extremely high including agreement (99.2%, sensitivity (100.00%, specificity (99.70%, positive (96.40% and negative predictive values (100.00% and accuracy (99.80%. The prevalence of STDs was very high (55.3%. Furthermore, associations were observed between STDs and changes in semen parameters, highlighting the importance of STD detection in semen. Thus, this M-PCR assay has great potential for application in semen screening programs for pathogens in infertility and STD clinics and in sperm banks.

  1. Whole blood Nested PCR and Real-time PCR amplification of Talaromyces marneffei specific DNA for diagnosis.

    Science.gov (United States)

    Lu, Sha; Li, Xiqing; Calderone, Richard; Zhang, Jing; Ma, Jianchi; Cai, Wenying; Xi, Liyan

    2016-02-01

    Talaromyces marneffei is a dimorphic pathogenic fungus, which is a life-threatening invasive mycosis in the immunocompromised host. Prompt diagnosis of T. marneffei infection remains difficult although there has been progress in attempts to expedite the diagnosis of this infection. We previously demonstrated the value of nested polymerase chain reaction (PCR) to detect T. marneffei in paraffin embedded tissue samples with high sensitivity and specificity. In this study, this assay was used to detect the DNA of T. marneffei in whole blood samples. Real-time PCR assay was also evaluated to identify T. marneffei in the same samples. Twenty out of 30 whole blood samples (67%) collected from 23 patients were found positive by using the nested PCR assay, while 23/30 (77%) samples were found positive by using the real-time PCR assay. In order to express accurately the fungal loads, we used a normalized linearized plasmid as an internal control for real-time PCR. The assay results were correlated as the initial quantity (copies/μl) with fungal burden. These data indicate that combination of nested PCR and real-time PCR assay provides an attractive alternative for identification of T. marneffei DNA in whole blood samples of HIV-infected patients. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Detection of Coxiella burnetii in ticks by PCR and by PCR - Restriction Fragment Length Polymorphism (RFLP)

    International Nuclear Information System (INIS)

    2010-01-01

    Coxiella burnetii, as an obligata intracellular bacterium, is the etiologic agent of Q-fever. It is widely distributed in nature and is responsible for infection in various animals (cattle, sheep, goat) and humans. C. burnetii has been isolated from milk, ticks and human patients with acute and chronic Q fever. Ticks are the principal vectors and reservoirs of C. burnetii. Since over 40 species of ticks have been found to be infected with C. burnetii, ticks can serve as indicators of infection in nature. In this study, total of 2472 ticks (1446 female, 1021 male and 5 nymphs) were collected from 38 provinces of Turkey. The ticks were gathered into groups of 1 to 7 ticks as to the provinces, species and gender for DNA extraction. Following DNA extraction, the groups were examined for the presence of C. burtii by using the CB1and CB2. The ticks collected from the province of Denizli (56 in total) were gathered into 13 groups according to the species and gender. From these groups, 6 were positive for C. burnetii. The ticks collected from Ankara province, total of 160 ticks, were grouped into 53 as to their species and gender, only one group was found to be positive for C. burnetii. The specificities of PCR products were evaluated by restriction analysis. The positive PCR products were digested with the enzyme Taq1 and for bands in order of 118, 57, 43 and 39 bp's were appeared such as seen in the positive control DNA (C. burnetii Nine Mile RSA493)

  3. DESULFURACIÓN DE RELAVE MEDIANTE LA FLOTACIÓN DE SULFUROS DE HIERRO

    Directory of Open Access Journals (Sweden)

    Luis Valderrama Campusano

    2015-12-01

    Full Text Available En Chile la minería se desarrolla principalmente en  la concentración de cobre, molibdeno, oro y plata, mediante el proceso de flotación. Este genera grandes tonelajes de relaves que contienen diversas minerales, principalmente pirita. Dado que estos minerales se depositan en los tranques, estos pueden generar aguas ácidas; se propone como alternativa la desulfuración de los súlfuros por flotación. En una primera parte se estudió la flotabilidad de la pirita en un tubo Hallimond, utilizando como colector AP 404. Luego se realizaron pruebas de flotación a nivel de laboratorio en celda Denver D-12, utilizando pulpa de relave fresco de la etapa rougher de un circuito de flotación de cobre,  determinando el pH óptimo y dosificación de colector y espumantes. Pruebas de cinética de flotación permitieron determinar el tiempo óptimo de flotación tanto para la etapa rougher como  para la etapa scavenger. Se concluye que la flotación de los relaves sea una técnicamente factible su desulfuración,  obteniendo relaves que contienen  0,08% de pirita.

  4. Interoperabilidad en Sistemas Domóticos Mediante Pasarela Infrarrojos-ZigBee

    Directory of Open Access Journals (Sweden)

    Gonzalo B. Asencio

    2011-10-01

    Full Text Available Resumen: La domótica consiste en la aplicación de técnicas provenientes de la automática industrial al hogar con objeto de ofrecer servicios que aporten, entre otras cosas, confort, seguridad y eficiencia energética a los usuarios. Hasta el momento la penetración de dichas técnicas en los hogares ha sido reducida. Una de las razones fundamentales de esta lenta transposición de técnicas de control al hogar es la dificultad de integración entre los diferentes sistemas presentes en el hogar. En este artículo se presenta un desarrollo encaminado a mejorar la integración de los sistemas domóticos con aquellos dispositivos que sean controlables mediante infrarrojos. En concreto se ha desarrollado una pasarela inalámbrica que permite a una red domótica el envío de tramas de infrarrojos. De esta manera se posibilita un despliegue rápido y económico de los nodos que sean necesarios para integrar dispositivos tales como los sistemas de aire acondicionado en una red domótica. Copyright c 2011 CEA. Publicado por Elsevier España, S.L. Todos los derechos reservados. Palabras clave: Control a través de redes de comunicación, Impacto social de la automática

  5. Galactosemia: Diagnóstico precoz mediante estudio enzimático

    Directory of Open Access Journals (Sweden)

    Úrsula Carrillo Estrada

    2003-09-01

    Full Text Available Se presentan los resultados obtenidos del estudio realizado a un paciente masculino de 45 días de nacido, cuyo motivo de ingreso fue pérdida de peso, decaimiento, retraso psicomotor y crisis de hipoglucemia. Los síntomas comenzaron en el período neonatal y coincidieron con la introducción de la lactancia materna. En estudios realizados se constató en la orina la presencia de lactosa y galactosa. Se confirma el diagnóstico por estudio enzimático. La evolución clínica ha sido satisfactoria. El tratamiento dietético que excluía a los alimentos que contienen galactosa y lactosa fue de mucha importancia. Es el primer caso diagnosticado en Cuba mediante estudio enzimático.This paper presents the results of a study performed on a 45-day old male patient who was admitted to the hospital for weight loss, tiredness, psychomotor retardation and hypoglicemic crisis. The symptoms had begun in the neonatal period and had coincided with the introduction of breast feeding. The studies detected lactose and galactose in urine. The enzymatic study confirmed the diagnosis. The clinical recovery was satisfactory. The dietary treatment that excluded foods containing galactose and lactose was important and successful. He is the first case diagnosed on enzymatic study in Cuba.

  6. Prácticas educativas. Desarrollo de la lectura mediante estrategias integradoras

    Directory of Open Access Journals (Sweden)

    Maira Solé

    2005-01-01

    Full Text Available La lectura y la escritura son procesos que cada día ameritan nuevos cambios y transformaciones. La propuesta de un Proyecto Pedagógico Integrador, (Fraca 2003 desarrollado con éxito en algunas instituciones venezolanas, se perfila como una alternativa significativa para el desarrollo de estos elementos. La idea o núcleo central es la integración de las diferentes asignaturas curriculares y lograr una globalización partiendo de sus objetivos y contenidos programáticos. El eje pedagógico integrador le permite al docente, evidenciar con mayor prontitud los resultados mediante actividades prácticas de lectura y escritura. Así mismo combina elementos claves del aprendizaje ausbeliano: información previa, información nueva y construcción de la información definitiva o integrada. La puesta en ejecución de las estrategias integradoras, en esta ocasión por maestros en formación (UNEG, a diferentes niños de escuelas del Estado Bolívar (Venezuela, certificando cómo la lectura y la escritura pueden tener un espacio ideal y significativo en la instrucción actual. Solo se necesita la intención, creatividad, dinamismo e ingenio.

  7. BIORREMEDIACIÓN DE UN SUELO CON DIESEL MEDIANTE EL USO DE MICROORGANISMOS AUTÓCTONOS

    Directory of Open Access Journals (Sweden)

    ARRIETA RAMÍREZ OLGA MARIA

    2012-07-01

    Full Text Available En este estudio, se aisló y caracterizó bioquímica y molecularmente un consorcio bacteriano capaz de degradar los diferentes hidrocarburos presentes en un combustible diesel,conformado por los siguientes géneros: Enterobacter sp, Bacillus sp, Staphylococcus aureus, Sanguibacter soli, Arthrobacter sp y Flavobacterium sp, a partir de un suelo contaminado con diesel a escala de laboratorio, y tratado mediante 2 tecnologías de biorremediación: atenuación natural y bioestimulación. Se definió como parámetro de control la concentración de Hidrocarburos Totales del Petróleo (HTP y para el cual, se obtuvo una reducción en la concentración en un periodo de 4 meses de 36,86% para atenuación natural y 50,99% para bioestimulación. La medición de la eficiencia de remoción de hidrocarburos se cuantificó por cromatografía de gases acoplada a masas (GC-MS.

  8. Análisis de una columna desbutanizadora mediante técnica de perfilaje gamma

    Directory of Open Access Journals (Sweden)

    Óscar Chaverri

    2007-05-01

    Full Text Available En este trabajo se reportan los resultados de un estudio, realizado mediante la técnica no destructiva de perfilaje por rayos gamma, de una columna desbutanizadora en una refinería de petróleo. Dicho estudio tuvo entre sus objetivos el de resaltar los beneficios que se derivan de la aplicación pacifica de la energía nuclear, en la determinación de problemas que afectan la eficiencia y seguridad de procesos industriales. En este caso particular, se inspeccionó parte de una torre para la destilación de petróleo, y se determinó la presencia de algunas pequeñas anomalías, aunque el funcionamiento de la columna era correcto y seguro. Cabe recalcar que,por tratarse de una técnica no destructiva y con la posibilidad de medir a través de la pared de la columna, el ensayo fue realizado sin necesidad de abrir la torre misma o de introducir personal técnico en su interior.

  9. Operación de un PLC Mediante un PDA Vía ZIGBEE

    Directory of Open Access Journals (Sweden)

    Juan Zuluaga-Botero

    2015-07-01

    Full Text Available En este artículo se muestra la parte preliminar de interconexión de un módulo ZigBee con un Asistente Digital Personal (PDA a través de la red del Sistema de telefonía Móvil Global (GSM, para aplicación futura de comunicación con un Controlador Lógico Programable (PLC. Esta aplicación permite realizar monitoreo y control del sistema, de manera remota y con dispositivos móviles siendo el alcance del proyecto de investigación. Para este proyecto inicialmente se realizan comunicaciones que permiten empalmar las tecnologías de las redes de telefonía móvil celular con la red ZigBee, haciendo una interfaz transparente para el usuario, presentando dichos resultados en este artículo. Para esto se trabaja con tramas de datos básicas a través de la red de telefonía móvil celular, mediante mensajería corta recibida por el puerto serial del Zigbee remoto, utilizando microcontroladores para el control de los módulos que conforman esta parte del sistema de comunicación, estableciendo la interfaz con el dispositivo móvil o Smartphone vía Zigbee.

  10. Medida de similaridad entre imágenes de marcas de ganado mediante distribuciones de forma

    Directory of Open Access Journals (Sweden)

    Germán Sánchez Torres

    2014-12-01

    Full Text Available Este documento reporta los resultados de una investigación orientada hacia el diseño de un método de tratamiento de imágenes digitales para la automatización de los procesos de registro y control de marcas de ganado requeridas por las regulaciones del sector ganadero en Colombia. El método permite automatizar los procesos de búsqueda y de comparación necesarios para garantizar la unicidad de las marcas dentro de un sistema asistido por computadora. Se inicia con la generación de un histograma estimado de la geometría de la marca, lo que permite comparar y detectar similitudes entre las figuras previamente almacenadas, mediante una métrica de similitud basada en la distancia de Minkowski. Los resultados obtenidos indican que el método es adecuado para realizar un proceso de discriminación de dichas imágenes, reducir las ambigüedades y garantizar la unicidad de los registros. Los resultados obtenidos y un análisis de su aplicación son reportados.

  11. Vigilancia del riesgo de ocurrencia de incendios forestales mediante estaciones meteorológicas de superficie

    Directory of Open Access Journals (Sweden)

    I. M. Domínguez-Hurtado

    2008-01-01

    Full Text Available Para la estimación del riesgo de ocurrencia de incendios forestales se emplean diferentes variantes a nivel mundial, en este trabajo se propone un sistema de vigilancia para la detección del riesgo de ocurrencia de incendios forestales a partir de la información procedente de estaciones meteorológicas de superficie, debido básicamente a la carencia de salidas operativas diarias que evaluarán las condiciones de riesgo de fuegos a nivel nacional. Se emplearon los índices de Nesterov modificado y Monte Alegre, los cuales se estimaron a partir de los datos procedentes de las 68 estaciones meteorológicas de Cuba. A partir de dicha información se obtiene la salida mapificada diaria de cada uno de ellos, mediante la utilización de la técnica de interpolación bicubic spline. Se obtuvo un sistema operativo totalmente automatizado para la evaluación de las condiciones de riesgo de ocurrencia de incendios para Cuba, el cual brinda la posibilidad de completar y perfeccionar el sistema de vigilancia de fuegos, basado en la reducción de la escala temporal y el tratamiento de la información.

  12. LA EXPERIENCIA DE UNA CLASE INTEGRADA MEDIANTE RESOLUCIÓN DE PROBLEMAS

    Directory of Open Access Journals (Sweden)

    Fernanda Ortiz Rivera

    2014-05-01

    Full Text Available Es una experiencia en la Institución educativa Tomas Cipriano de Mosquera con estudiantes del ciclo noveno en una clase de ciencias naturales sobre el sentido de la audición, teniendo en cuenta integrar la biología  y la física  mediante el tema del órgano de la audición con el tema del sonido, en el marco de un enfoque didáctico por resolución de problemas, en el cual la pregunta problema central que se generó a los estudiantes, producto de consultar sus intereses, fue: ¿por qué se produce la sordera? Esta pregunta se realizó al comienzo y al final de la clase, mostrando de esta manera al final buenos resultados de aprendizaje en los estudiantes, ya que fueron construyendo  nociones, ideas y conceptos necesarios sobre el sonido con sus propiedades y cualidades; y la audición con las funciones y partes del oído, para poder resolver la pregunta central gracias a una secuencia didáctica de clase que construimos cuidadosamente en el que incluía experimentos, lecturas, explicaciones, materiales didácticos, entre otros, siendo así una clase muy activa y participativa, ya que el estudiante siempre fue el que llevo el papel principal de la clase.

  13. Determinación del umbral de detección de Pseudococcus viburni (Hemiptera: Pseudococcidae por PCR Determination of the detection threshold of Pseudococcus viburni (Hemiptera: Pseudococcidae by PCR

    Directory of Open Access Journals (Sweden)

    Diana Vera

    2012-06-01

    Full Text Available La cochinilla harinosa de los frutales Pseudococcus viburni (Signoret es una plaga cuarentenaria, presente en el Alto Valle de Río Negro y Neuquén, Argentina. Su detección durante la fiscalización aduanera, aun en los estados inmaduros, provoca rechazos de la fruta fresca argentina con destino a los mercados internacionales. Las técnicas actuales de identificación de pseudocóccidos y otros cocoideos implican la realización de preparados microscópicos que requieren varios días. Por esto, la disminución de los tiempos de identificación es importante sobre todo en las tareas de fiscalización. En este trabajo, se determinó el umbral de detección específica de P. viburni mediante PCR, como así también, la implementación de un método rápido de extracción de ADN mediante DNAzolT. Insectos de diferentes estados de desarrollo (huevo, ninfas (tres estados ninfales y adulto, conservados en etanol pro análisis a -20 ºC, provenientes de montes frutales del Alto Valle, Argentina, fueron procesados según el protocolo del fabricante y se logró obtener ADN de buena calidad y concentración. Una alícuota del mismo fue utilizado como templado para una reacción de PCR usando primers específicos para P. viburni, registrados en bibliografía y como control positivo ADN de P. viburni de colección entomológica. Los primers utilizados y sus secuencias son A4 (5'-cccgcggccgttctctcttt-3' y A5 (5'-atatgttgtgcatagttgtgtgtgcgc-3', diseñados por Beuning et al. (1999. La amplificación generó una banda de peso molecular esperado de 650 pb. en gel de agarosa al 1.5% en todos los estadios, se determinó como límite de detección el estado de huevo. Esta técnica constituye una detección específica de P. viburni en un lapso máximo de 48 h.The obscure mealybug Pseudococcus viburni (Signoret is a quarantine pest present in the Upper Valley of Río Negro and Neuquén, Argentina. The detection of any growth stage of the mealybug in quarantine

  14. DNA extraction method for PCR in mycorrhizal fungi.

    Science.gov (United States)

    Manian, S; Sreenivasaprasad, S; Mills, P R

    2001-10-01

    To develop a simple and rapid DNA extraction protocol for PCR in mycorrhizal fungi. The protocol combines the application of rapid freezing and boiling cycles and passage of the extracts through DNA purification columns. PCR amplifiable DNA was obtained from a number of endo- and ecto-mycorrhizal fungi using minute quantities of spores and mycelium, respectively. DNA extracted following the method, was used to successfully amplify regions of interest from high as well as low copy number genes. The amplicons were suitable for further downstream applications such as sequencing and PCR-RFLPs. The protocol described is simple, short and facilitates rapid isolation of PCR amplifiable genomic DNA from a large number of fungal isolates in a single day. The method requires only minute quantities of starting material and is suitable for mycorrhizal fungi as well as a range of other fungi.

  15. Some factors determining the PCr recovery overshoot in skeletal muscle.

    Science.gov (United States)

    Korzeniewski, Bernard; Zoladz, Jerzy A

    2005-07-01

    It has been proposed recently that the phosphocreatine (PCr) overshoot (increase above the resting level) during muscle recovery after exercise is caused by a slow decay during this recovery of the direct activation of oxidative phosphorylation taking place during muscle work. In the present article the factors determining the appearance and size of the PCr overshoot are studied using the computer model of oxidative phosphorylation in intact skeletal muscle developed previously. It is demonstrated that the appearance and duration of this overshoot is positively correlated with the value of the characteristic decay time of the direct activation of oxidative phosphorylation. It is also shown that the size of PCr overshoot is increased by low resting PCr/Cr ratio (what is confirmed by our unpublished experimental data), by high intensity of the direct activation of oxidative phosphorylation, by high muscle work intensity and by low rate of the return of cytosolic pH to the resting value during muscle recovery.

  16. polymerase chain reaction (pcr) provides a superior tool

    African Journals Online (AJOL)

    boaz

    Keywords: Streptococcus pneumoniae, meningitis, rt-PCR, standard bacteriological methods ... qualification de techniciens et les matériels pour son application constituent des ... Streptococcus pneumonia (pneumococcus) is a common.

  17. Multiplex polymerase chain reaction (PCR) and fluorescence-based ...

    African Journals Online (AJOL)

    reading 7

    2011-12-28

    Dec 28, 2011 ... mitochondrial DNA and cytochrome b as an internal PCR control. The amplified species- ... more labour-saving than using each pair of species- specific primers separately for .... obtained from the NCBI nucleotide data bank.

  18. Comparison of PCR-ELISA and LightCycler real-time PCR assays for detecting Salmonella spp. in milk and meat samples

    DEFF Research Database (Denmark)

    Perelle, Sylvie; Dilasser, Françoise; Malorny, Burkhard

    2004-01-01

    , minced beef and raw milk, and 92 naturally-contaminated milk and meat samples. When using either PCR-ELISA or LC-PCR assays, only Salmonella strains were detected. PCR-ELISA and LC-PCR assays gave with pure Salmonella cultures the same detection limit level of 10(3) CFU/ml, which corresponds respectively...

  19. Development of RT-PCR and Nested PCR for Detecting Four Quarantine Plant Viruses Belonging to Nepovirus

    Directory of Open Access Journals (Sweden)

    Siwon Lee

    2013-09-01

    Full Text Available For quarantine purpose, we developed the RT- and nested PCR module of Tomato black ring virus (TBRV, Arabis mosaic virus (ArMV, Cherry leafroll virus (CLRV and Grapevine fanleaf virus (GFLV. The PCR modules, developed in this study make diagnosis more convenient and speedy because of same PCR condition. And also, the methods are more accurate because it can check whether the result is contamination or not using the mutation-positive control. We discard or return the 27 cases of Nepovirus infection seed by employing the module past 3 years. This study provides a rapid and useful method for detection of four quarantine plant viruses.

  20. Procedimiento de estabilización de mercurio líquido mediante cemento polimérico de azufre, vía sulfuro de mercurio.

    OpenAIRE

    López-Delgado, Aurora; López Gómez, Félix Antonio; Alguacil, Francisco José; Alonso Gámez, Manuel

    2011-01-01

    Procedimiento de estabilización de mercurio líquido mediante cemento polimérico de azufre, vía sulfuro de mercurio. Procedimiento para la estabilización de mercurio líquido mediante la obtención de cementos poliméricos de azufre que comprende: (a) transformación del mercurio líquido en sulfuro de mercurio (metacinabrio) mediante reacción química, en condiciones estequiométricas, entre el mercurio y el azufre elemental; y (b) obtención de cemento polimérico de azufre me...

  1. Diseño mediante elementos finitos de componentes estructurales de un cuadricóptero para impresión 3D

    OpenAIRE

    PARDO APARISI, IVÁN

    2016-01-01

    El trabajo tiene como objetivo diseñar componentes estructurales, mediante el método de elementos finitos, que serán utilizados en un dron de cuatro rotores (cuadricóptero). Una característica particular de este proyecto es que los componentes estructurales a diseñar serán fabricados mediante impresión 3D. Pardo Aparisi, I. (2016). Diseño mediante elementos finitos de componentes estructurales de un cuadricóptero para impresión 3D. http://hdl.handle.net/10251/75994. TFGM

  2. Culture independent PCR: an alternative enzyme discovery strategy

    DEFF Research Database (Denmark)

    Jacobsen, Jonas; Lydolph, Magnus; Lange, Lene

    2005-01-01

    Degenerate primers were designed for use in a culture-independent PCR screening of DNA from composite fungal communities, inhabiting residues of corn stovers and leaves. According to similarity searches and alignments amplified clone sequences affiliated with glycosyl hydrolase family 7 and glyco...... the value of culture-independent PCR in microbial diversity studies and could add to development of a new enzyme screening technology....

  3. MPprimer: a program for reliable multiplex PCR primer design

    Directory of Open Access Journals (Sweden)

    Wang Xiaolei

    2010-03-01

    Full Text Available Abstract Background Multiplex PCR, defined as the simultaneous amplification of multiple regions of a DNA template or multiple DNA templates using more than one primer set (comprising a forward primer and a reverse primer in one tube, has been widely used in diagnostic applications of clinical and environmental microbiology studies. However, primer design for multiplex PCR is still a challenging problem and several factors need to be considered. These problems include mis-priming due to nonspecific binding to non-target DNA templates, primer dimerization, and the inability to separate and purify DNA amplicons with similar electrophoretic mobility. Results A program named MPprimer was developed to help users for reliable multiplex PCR primer design. It employs the widely used primer design program Primer3 and the primer specificity evaluation program MFEprimer to design and evaluate the candidate primers based on genomic or transcript DNA database, followed by careful examination to avoid primer dimerization. The graph-expanding algorithm derived from the greedy algorithm was used to determine the optimal primer set combinations (PSCs for multiplex PCR assay. In addition, MPprimer provides a virtual electrophotogram to help users choose the best PSC. The experimental validation from 2× to 5× plex PCR demonstrates the reliability of MPprimer. As another example, MPprimer is able to design the multiplex PCR primers for DMD (dystrophin gene which caused Duchenne Muscular Dystrophy, which has 79 exons, for 20×, 20×, 20×, 14×, and 5× plex PCR reactions in five tubes to detect underlying exon deletions. Conclusions MPprimer is a valuable tool for designing specific, non-dimerizing primer set combinations with constrained amplicons size for multiplex PCR assays.

  4. Template preparation for rapid PCR in Colletotrichum lindemuthianum

    Directory of Open Access Journals (Sweden)

    Roca M. Gabriela

    2003-01-01

    Full Text Available Isolation of DNA for PCR is time-consuming and involves many reagents. The aim of this work was to optimise a rapid and easy PCR methodology without previous DNA isolation. Different strains of the phytopathogenic fungus Colletotrichum lindemuthianum were used. Protoplasts were generated using lytic enzymes under high incubation temperatures using different methodologies to obtain the template. A rapid (10 minute methodology was successful for smaller amplicons (<750 bp.

  5. DNA microarray-based PCR ribotyping of Clostridium difficile.

    Science.gov (United States)

    Schneeberg, Alexander; Ehricht, Ralf; Slickers, Peter; Baier, Vico; Neubauer, Heinrich; Zimmermann, Stefan; Rabold, Denise; Lübke-Becker, Antina; Seyboldt, Christian

    2015-02-01

    This study presents a DNA microarray-based assay for fast and simple PCR ribotyping of Clostridium difficile strains. Hybridization probes were designed to query the modularly structured intergenic spacer region (ISR), which is also the template for conventional and PCR ribotyping with subsequent capillary gel electrophoresis (seq-PCR) ribotyping. The probes were derived from sequences available in GenBank as well as from theoretical ISR module combinations. A database of reference hybridization patterns was set up from a collection of 142 well-characterized C. difficile isolates representing 48 seq-PCR ribotypes. The reference hybridization patterns calculated by the arithmetic mean were compared using a similarity matrix analysis. The 48 investigated seq-PCR ribotypes revealed 27 array profiles that were clearly distinguishable. The most frequent human-pathogenic ribotypes 001, 014/020, 027, and 078/126 were discriminated by the microarray. C. difficile strains related to 078/126 (033, 045/FLI01, 078, 126, 126/FLI01, 413, 413/FLI01, 598, 620, 652, and 660) and 014/020 (014, 020, and 449) showed similar hybridization patterns, confirming their genetic relatedness, which was previously reported. A panel of 50 C. difficile field isolates was tested by seq-PCR ribotyping and the DNA microarray-based assay in parallel. Taking into account that the current version of the microarray does not discriminate some closely related seq-PCR ribotypes, all isolates were typed correctly. Moreover, seq-PCR ribotypes without reference profiles available in the database (ribotype 009 and 5 new types) were correctly recognized as new ribotypes, confirming the performance and expansion potential of the microarray. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  6. Diagnosis of aerobic vaginitis by quantitative real-time PCR

    OpenAIRE

    Rumyantseva, T. A.; Bellen, G.; Savochkina, Y. A.; Guschin, A. E.; Donders, G.G.G.

    2016-01-01

    Abstract: Purpose To evaluate a real-time PCR-based technique to quantify bacteria associated with aerobic vaginitis (AV) as a potential test. Methods Vaginal samples from 100 women were tested by wet-mount microscopy, gram stain and quantitative real-time PCR targeting Enterobacteriacea, Staphylococcus spp., Streptococcus spp., Enterococcus spp., Escherichia coli, Streptococcus agalactiae, S. aureus; Lactobacillus spp. AV diagnosis obtained by wet-mount microscopy was used as reference. Resu...

  7. [Optimized application of nested PCR method for detection of malaria].

    Science.gov (United States)

    Yao-Guang, Z; Li, J; Zhen-Yu, W; Li, C

    2017-04-28

    Objective To optimize the application of the nested PCR method for the detection of malaria according to the working practice, so as to improve the efficiency of malaria detection. Methods Premixing solution of PCR, internal primers for further amplification and new designed primers that aimed at two Plasmodium ovale subspecies were employed to optimize the reaction system, reaction condition and specific primers of P . ovale on basis of routine nested PCR. Then the specificity and the sensitivity of the optimized method were analyzed. The positive blood samples and examination samples of malaria were detected by the routine nested PCR and the optimized method simultaneously, and the detection results were compared and analyzed. Results The optimized method showed good specificity, and its sensitivity could reach the pg to fg level. The two methods were used to detect the same positive malarial blood samples simultaneously, the results indicated that the PCR products of the two methods had no significant difference, but the non-specific amplification reduced obviously and the detection rates of P . ovale subspecies improved, as well as the total specificity also increased through the use of the optimized method. The actual detection results of 111 cases of malarial blood samples showed that the sensitivity and specificity of the routine nested PCR were 94.57% and 86.96%, respectively, and those of the optimized method were both 93.48%, and there was no statistically significant difference between the two methods in the sensitivity ( P > 0.05), but there was a statistically significant difference between the two methods in the specificity ( P PCR can improve the specificity without reducing the sensitivity on the basis of the routine nested PCR, it also can save the cost and increase the efficiency of malaria detection as less experiment links.

  8. Diagnóstico de distrofia muscular de Duchenne mediante análisis del ácido desoxinucleotico y su aplicación en la prevención

    Directory of Open Access Journals (Sweden)

    Mayra Rodríguez Hernández

    1996-04-01

    Full Text Available Se define una estrategia para la prevención en Cuba de la distrofia muscular de Duchenne (DMD, una de las enfermedades hereditarias letales más frecuentes, y se evalúan la factibilidad de su aplicación y los problemas que pudieran dificultar su implantación al nivel nacional. La estrategia se basa fundamentalmente en la necesidad de detectar las familias afectadas, la definición de las mujeres portadoras o en riesgo de serlo y el estudio molecular de los miembros de interés con anterioridad al ofrecimiento de los servicios de diagnóstico prenatal mediante análisis directo -detección de deleciones en el gen DMD mediante reacción en cadena de la polimerasa (PCR o análisis indirecto- empleo de los marcadores denominados polimorfismos en la longitud de los fragmentos de restricción (RFLPs en análisis de ligamento. Se concluye en que la aplicación de esta estrategia es factible y conveniente, pues permite ofrecer el diagnóstico prenatal al 75 % de las mujeres portadoras. Su eficiencia en la prevención del nacimiento de nuevos enfermos DMD se demuestra en 2 diagnósticos prenatales realizados, uno de los cuales detectó un embarazo afectado que fue interrumpido por solicitud de los padres.A strategy is defined for the prevention in Cuba of the Duchenne's muscular dystrophy (DMD, one of the most frequent lethal hereditary diseases, and the feasibility of its application, and the troubles that might difficult its implantation at a national level, are evaluated. This strategy is mainly based on the need of detecting the affected families, the definition of the carrier women, or the women at risk of being carriers, and the molecular study of the members of interest with anteriority to the offering of prenatal diagnosis services by direct analysis -detection of DMD gen deletions by (PCR polymerase chain reaction, or indirect analysis-, use of the markers called polymorphisms in the length of the restriction fragments (RFLPs in ligament

  9. Diseño y prototipaje del álabe para un miniaerogenerador mediante impresión 3D

    OpenAIRE

    Roy Mota, Andrea

    2017-01-01

    El objetivo de este proyecto consiste en el diseño de una maqueta de álabe para un mini aerogenerador y su posterior fabricación con PLA mediante la tecnología de impresión 3D no industrial. Para conseguirlo se creó una hoja de cálculo que torna la superficie del ala; se analizó la impresora 3D y se diseñó la estructura interna del aspa para dotarlo de resistencia según sus límites de impresión de la impresora mediante el programa Siemens Unigraphics NX10; se simularon los esfuerzos y a parti...

  10. Extracción de ADN de Trypanosoma cruzi mediante tratamiento con bromuro de hexadecil-trimetil-amonio

    Directory of Open Access Journals (Sweden)

    Marcela Escalante

    1997-06-01

    Full Text Available En el presente trabajo se describe un método rápido, sencillo y eficaz para la obtención de ADN genómico de Trypanosoma cruzi, libre de impurezas y fácil de manipular. Dicho procedimiento se basa en la lisis del parásito con SDS y remoción de proteínas mediante la digestión con proteinasa K, seguida de la precipitación selectiva de carbohidratos y proteínas residuales con bromuro de hexadecil-trimetil-amonio (CTAB. Finalmente, el ADN se extrae con cloroformo: alcohol isoamílico y se recupera de la fase acuosa mediante precipitación con isopropanol.

  11. Control mediante modos deslizantes en tiempo discreto para el seguimiento de trayectorias de un robot móvil1

    Directory of Open Access Journals (Sweden)

    P.A. Niño-Suárez

    2007-10-01

    Full Text Available Resumen: En este trabajo se presenta una estrategia de control en tiempo discreto para el seguimiento de trayectorias de un robot móvil tipo (2,0 controlado remotamente. La estrategia de control se desarrolló mediante un enfoque de modos deslizantes, considerando el modelo discreto exacto del vehículo en el cual se incluyen los efectos del retardo de transporte causado por la propagación de las señales sobre una red de comunicación. El esquema de control garantiza el seguimiento de trayectorias predeterminadas obteniéndose convergencia asintótica de los errores de seguimiento. La estrategia propuesta es evaluada mediante una serie de resultados por simulación. Palabras clave: Robot móvil, retardos de transporte, control en tiempo discreto, modos deslizantes

  12. PCR performance of a thermostable heterodimeric archaeal DNA polymerase

    Science.gov (United States)

    Killelea, Tom; Ralec, Céline; Bossé, Audrey; Henneke, Ghislaine

    2014-01-01

    DNA polymerases are versatile tools used in numerous important molecular biological core technologies like the ubiquitous polymerase chain reaction (PCR), cDNA cloning, genome sequencing, and nucleic acid based diagnostics. Taking into account the multiple DNA amplification techniques in use, different DNA polymerases must be optimized for each type of application. One of the current tendencies is to reengineer or to discover new DNA polymerases with increased performance and broadened substrate spectra. At present, there is a great demand for such enzymes in applications, e.g., forensics or paleogenomics. Current major limitations hinge on the inability of conventional PCR enzymes, such as Taq, to amplify degraded or low amounts of template DNA. Besides, a wide range of PCR inhibitors can also impede reactions of nucleic acid amplification. Here we looked at the PCR performances of the proof-reading D-type DNA polymerase from P. abyssi, Pab-polD. Fragments, 3 kilobases in length, were specifically PCR-amplified in its optimized reaction buffer. Pab-polD showed not only a greater resistance to high denaturation temperatures than Taq during cycling, but also a superior tolerance to the presence of potential inhibitors. Proficient proof-reading Pab-polD enzyme could also extend a primer containing up to two mismatches at the 3' primer termini. Overall, we found valuable biochemical properties in Pab-polD compared to the conventional Taq, which makes the enzyme ideally suited for cutting-edge PCR-applications. PMID:24847315

  13. PCR performance of a thermostable heterodimeric archaeal DNA polymerase

    Directory of Open Access Journals (Sweden)

    Tom eKillelea

    2014-05-01

    Full Text Available DNA polymerases are versatile tools used in numerous important molecular biological core technologies like the ubiquitous polymerase chain reaction (PCR, cDNA cloning, genome sequencing and nucleic acid based diagnostics. Taking into account the multiple DNA amplification techniques in use, different DNA polymerases must be optimized for each type of application. One of the current tendencies is to reengineer or to discover new DNA polymerases with increased performance and broadened substrate spectra. At present, there is a great demand for such enzymes in applications, e.g., forensics or paleogenomics. Current major limitations hinge on the inability of conventional PCR enzymes, such as Taq, to amplify degraded or low amounts of template DNA. Besides, a wide range of PCR inhibitors can also impede reactions of nucleic acid amplification. Here we looked at the PCR performances of the proof-reading D-type DNA polymerase from P. abyssi, Pab-polD. Fragments, 3 kilobases in length, were specifically PCR-amplified in its optimized reaction buffer. Pab-polD showed not only a greater resistance to high denaturation temperatures than Taq during cycling, but also a superior tolerance to the presence of potential inhibitors. Proficient proof-reading Pab-polD enzyme could also extend a primer containing up to two mismatches at the 3’ primer termini. Overall, we found valuable biochemical properties in Pab-polD compared to the conventional Taq, which makes the enzyme ideally suited for cutting-edge PCR-applications.

  14. Immunomagnetic nanoparticle based quantitative PCR for rapid detection of Salmonella

    International Nuclear Information System (INIS)

    Bakthavathsalam, Padmavathy; Rajendran, Vinoth Kumar; Saran, Uttara; Chatterjee, Suvro; Ali, Baquir Mohammed Jaffar

    2013-01-01

    We have developed a rapid and sensitive method for immunomagnetic separation (IMS) of Salmonella along with their real time detection via PCR. Silica-coated magnetic nanoparticles were functionalized with carboxy groups to which anti-Salmonella antibody raised against heat-inactivated whole cells of Salmonella were covalently attached. The immuno-captured target cells were detected in beverages like milk and lemon juice by multiplex PCR and real time PCR with a detection limit of 10 4 cfu.mL −1 and 10 3 cfu.mL −1 , respectively. We demonstrate that IMS can be used for selective concentration of target bacteria from beverages for subsequent use in PCR detection. PCR also enables differentiation of Salmonella typhi and Salmonella paratyphi A using a set of four specific primers. In addition, IMS—PCR can be used as a screening tool in the food and beverage industry for the detection of Salmonella within 3–4 h which compares favorably to the time of several days that is needed in case of conventional detection based on culture and biochemical methods. (author)

  15. Proyectar con la naturaleza mediante la Metodología de los Estudios de Impacto Ambiental en ordenaciones residenciales

    OpenAIRE

    Higueras García, Esther

    2013-01-01

    Proyectar con la naturaleza mediante la Metodología de los Estudios de Impacto Ambiental en ordenaciones residenciales .- Procedimiento de acción en la planificación ambiental .- Las técnicas de agregación de impactos .- Las afecciones de los planes de ordenación sobre el territorio. .- Las medidas preventivas y correctoras de planes .- Evaluación critica de los estudios de impacto ambiental

  16. Superficie específica de una bentonita mediante la adsorción de azul de metileno

    Directory of Open Access Journals (Sweden)

    Jorge Alejo Pinzón Bello

    2010-07-01

    Full Text Available Se estudió la determinación de la superficie específica de una bentonita colombiana, procedente del Valle del Cauca, mediante la adsorción de azul de metileno, a 298 K. Este método se comparó con el de la adsorción de nitrógeno a 77 K.

  17. Simulación del perfil tensión-corriente para paneles solares mediante convertidor CC-CC reductor

    OpenAIRE

    Muñoz Palaguachi, Henry Patricio; Patiño Guiracocha, Claudio Vinicio

    2017-01-01

    El desarrollo de este proyecto tiene como objetivo la reconstrucción del perfil tensión-corriente para paneles solares ante diferentes condiciones de funcionamiento al variar la temperatura e irradiancia. Este perfil se logra con el control de un convertidor reductor de tipo conmutado mediante controladores de corriente y tensión. In this work the emulation of voltage-current profiles for photovoltaic panels is developed for different operating conditions, specifically for variations in ir...

  18. IDENTIFIKASI DAGING BABI MENGGUNAKAN METODE PCR-RFLP GEN Cytochrome b DAN PCR PRIMER SPESIFIK GEN AMELOGENIN (Pork Identification Using PCR-RFLP of Cytochrome b Gene and Species Specific PCR of Amelogenin Gene

    Directory of Open Access Journals (Sweden)

    Yuny Erwanto

    2013-03-01

    Full Text Available A polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP and species specific PCR methods had been applied for identifying pork in mixture of meat. Pork sample in various levels (1, 3, 5 and 10% was prepared in mixture with beef, chicken and mutton. The primary CYTb1 and CYTb2 were designed in the mitochondrial cytochrome b b (cytochrome b gene and PCR successfully amplified fragments of 359 bp. To distinguish pig species existence, the amplified PCR products of mitochondrial DNA were cut by BseDI restriction enzyme. The result showed that pig mitochondrial DNA was cut into 131 and 228 bp fragments. A polymerase chain reaction (PCR method based on the nucleotide sequence variation in the amelogenin gene has been chosen for the specific identification of pork DNAs in mixture meat. The primers designed generated specific fragments of 353 and 312 bp length for pork. The specificity of the primary designed was tested on 4 animal species including pig, cattle, chicken and goat species. Analysis of experimental mixture meat demonstrated that 1% of raw pork tissues could be detected using PCR-RFLP with BseDI restriction enzyme but detection using species-specific PCR showed the cross reactivity to beef, chicken and mutton. The cytochrome b PCR-RFLP species identification assay yielded excellent results for identification of pig species. PCR-RFLP is a potentially reliable technique for detection of the existence of pork in animal food product for Halal authentication. Keywords: Pork identification, cytochrome b, amelogenin, polymerase chain reaction   ABSTRAK   Penelitian ini dilakukan untuk mengaplikasikan metode deteksi daging babi dalam campuan daging dengan sapi, kambing dan ayam melalui PCR-RFLP dan PCR dengan primer spesifik untuk babi. Level kontaminasi daging babi dibuat sebesar 1, 3, 5 dan 10% dari total daging dalam campuran. Metode PCR-RFLP menggunakan sepasang primer yaitu gen cytochrome b dari mitokondria yang

  19. Detección por PCR de Colletotrichum lindemuthianum en cultivos y semillas de frijol en Antioquia, Colombia

    Directory of Open Access Journals (Sweden)

    Leonardo Martínez Pacheco

    2014-12-01

    Full Text Available Colletotrichum lindemuthianum, agente causal de la antracnosis del frijol, es uno de los patógenos más limitantes en la producción de este cultivo. La detección y correcta identificación de este hongo resulta fundamental para el manejo de la enfermedad, siendo las pruebas moleculares alternativas rápidas y sensibles para este fin. Mediante la técnica de PCR se evaluaron cuatro juegos de cebadores (CY1/CY2, CD1/CD2, ClF4/ITS4 y ClF432/ClR533 para la detección de C. lindemuthianum a partir de tejidos foliares, de vainas y de semillas procedentes de cultivos de frijol de Antioquia, Colombia. Los resultados indicaron que el par CD1/CD2, dirigido al pseudogen de permeasa de hierro Ftr1, fue el más efectivo para detectar el hongo en tejidos y semillas de frijol, así como para identificar aislamientos en cultivos microbiológicos. Para los cebadores CY1/CY2, dirigidos a los ITS del rDNA, se recomienda un esquema de PCR-RFLPs con MseI (=Tru1I para la diferenciación con las especies C. orbiculare y C. trifolii. Estos cebadores generaron resultados consistentes cuando se utilizaron en combinación con ITS1 (ITS1/CY2 e ITS4 (CY1/ITS4. Finalmente, los cebadores ClF4/ITS4 resultaron en amplificaciones inespecíficas y ClF432/ClR533 en fragmentos de difícil resolución en electroforesis de agarosa. Este estudio servirá de apoyo para los programas de certificación de semilla y mejoramiento genético de frijol.

  20. Comparative validation using quantitative real-time PCR (qPCR and conventional PCR of bovine semen centrifuged in continuous density gradient

    Directory of Open Access Journals (Sweden)

    M.V. Resende

    2011-06-01

    Full Text Available The objective of the present study was to determine the sperm enrichment with X-bearing spermatozoa, after one centrifugation in a Percoll or OptiPrep continuous density gradient, using quantitative real-time polymerase chain reaction (qPCR of sperm DNA and resultant in vitro-produced bovine embryos by PCR. Frozen/thawed sperm was layered on density gradients and the tubes were centrifuged. Supernatants were gently aspirated and the sperm recovered from the bottom of the tubes. Cleavage and blastocyst rates were determined through in vitro production of embryos and PCR was performed to identify the embryos' genetic sex. A difference in blastocyst rate was found in the Percoll treatment compared to OptiPrep (P<0.05. The percentage of female embryos in the Percoll and OptiPrep groups was 62.0% and 47.1%, respectively. These results were confirmed by qPCR of spermatozoa DNA and underestimation was seen only in the Percoll group. It was possible to sexing sperm using simple approach.

  1. RT-PCR Detection of HIV in Republic of Macedonia

    Directory of Open Access Journals (Sweden)

    Golubinka Bosevska

    2008-11-01

    Full Text Available The aim of the study was to detect HIV RNA in seropositive patients using RT-PCR method and thus, to establish PCR methodology in the routine laboratory works.The total of 33 examined persons were divided in two groups: 1 13 persons seropositive for HIV; and 2 20 healthy persons - randomly selected blood donors that made the case control group. The subjects age was between 25 and 52 years (average 38,5.ELFA test for combined detection of HIV p24 antigen and anti HIV-1 + 2 IgG and ELISA test for detection of antibodies against HIV-1 and HIV-2, were performed for each examined person. RNA from the whole blood was extracted using a commercial kit based on salt precipitation. Detection of HIV RNA was performed using RT-PCR kit. Following nested PCR, the product was separated by electrophoresis in 1,5 % agarose gel. The result was scored positive if the band of 210bp was visible regardless of intensity Measures of precaution were taken during all the steps of the work and HIV infected materials were disposed of accordingly.In the group of blood donors ELFA, ELISA and RT-PCR were negative. Assuming that prevalence of HIV infection is zero, the clinical specificity of RT-PCR is 100 %. The analytical specificity of RT-PCR method was tested against Hepatitis C and B, Human Papiloma Virus, Cytomegalovirus, Herpes Simplex Virus, Rubella Virus, Mycobacterium tuberculosis, Chlamydia trachomatis. None of these templates yielded amplicon. In the group of 13 seropositive persons, 33 samples were analyzed. HIV RNA was detected in 15 samples. ELISA and ELFA test were positive in all samples. Different aliquots of the samples were tested independently and showed the same results. After different periods of storing the RNA samples at -70°C, RT-PCR reaction was identical to the one performed initially. The obtained amplicons were maintained frozen at -20°C for a week and the subsequently performed electrophoresis was identical to the previous one. The reaction is

  2. Detection of Streptococcus pneumoniae in whole blood by PCR.

    Science.gov (United States)

    Zhang, Y; Isaacman, D J; Wadowsky, R M; Rydquist-White, J; Post, J C; Ehrlich, G D

    1995-03-01

    Streptococcus pneumoniae is a major cause of bacteremia in both children and adults. Currently, the diagnosis of pneumococcal bacteremia relies on the isolation and identification of the bacteria from blood cultures. We have developed a sensitive assay for the detection of S. pneumoniae in whole blood by the PCR. A specific primer-probe set (JM201 and JM202 primers with JM204 probe) designed from the penicillin-binding protein 2B gene was demonstrated to reproducibly detect between 10 and 100 fg of input purified S. pneumoniae DNA. This assay system was shown to be inclusive for all strains of S. pneumoniae evaluated, including 15 different serotypes and a battery of penicillin-resistant and -sensitive strains. The specificity of this PCR-based assay was demonstrated by its inability to support amplification from a series of human, bacterial, and yeast genomic DNAs. A general specimen preparation method which should be suitable for the purification of DNA from any pathogens in whole blood was developed. With this protocol it was possible to detect S. pneumoniae-specific DNA from whole blood specimens inoculated with as little as 4 CFU/ml. Copurified human blood DNA, ranging from 0 to 4.5 micrograms per PCR, did not affect the sensitivity of S. pneumoniae detection by PCR. A blinded clinical trial was used to compare the PCR-based assay with standard microbiological blood culture for the detection of S. pneumoniae bacteremia in 36 specimens obtained from pediatric patients seen in the emergency room of Children's Hospital of Pittsburgh. With culture as the "gold standard," the PCR-based assay had a sensitivity of 80% (4 of 5 culture-positive specimens were PCR positive) and a specificity of 84% (26 of 31 culture-negative specimens were PCR negative). However, three patients whose specimens were PCR positive and culture negative had histories suggestive of bacteremia, including recent positive blood cultures, treatment with antibiotics, cellulitis, and multiple

  3. Species Identification of Fox-, Mink-, Dog-, and Rabbit-Derived Ingredients by Multiplex PCR and Real-Time PCR Assay.

    Science.gov (United States)

    Wu, Qingqing; Xiang, Shengnan; Wang, Wenjun; Zhao, Jinyan; Xia, Jinhua; Zhen, Yueran; Liu, Bang

    2018-05-01

    Various detection methods have been developed to date for identification of animal species. New techniques based on PCR approach have raised the hope of developing better identification methods, which can overcome the limitations of the existing methods. PCR-based methods used the mitochondrial DNA (mtDNA) as well as nuclear DNA sequences. In this study, by targeting nuclear DNA, multiplex PCR and real-time PCR methods were developed to assist with qualitative and quantitative analysis. The multiplex PCR was found to simultaneously and effectively distinguish four species (fox, dog, mink, and rabbit) ingredients by the different sizes of electrophoretic bands: 480, 317, 220, and 209 bp. Real-time fluorescent PCR's amplification profiles and standard curves showed good quantitative measurement responses and linearity, as indicated by good repeatability and coefficient of determination R 2  > 0.99. The quantitative results of quaternary DNA mixtures including mink, fox, dog, and rabbit DNA are in line with our expectations: R.D. (relative deviation) varied between 1.98 and 12.23% and R.S.D. (relative standard deviation) varied between 3.06 and 11.51%, both of which are well within the acceptance criterion of ≤ 25%. Combining the two methods is suitable for the rapid identification and accurate quantification of fox-, dog-, mink-, and rabbit-derived ingredients in the animal products.

  4. Analysis of ELA-DQB exon 2 polymorphism in Argentine Creole horses by PCR-RFLP and PCR-SSCP.

    Science.gov (United States)

    Villegas-Castagnasso, E E; Díaz, S; Giovambattista, G; Dulout, F N; Peral-García, P

    2003-08-01

    The second exon of equine leucocyte antigen (ELA)-DQB genes was amplified from genomic DNA of 32 Argentine Creole horses by PCR. Amplified DNA was analysed by PCR-restriction fragment length polymorphism (RFLP) and PCR-single-strand conformation polymorphism (SSCP). The PCR-RFLP analysis revealed two HaeIII patterns, four RsaI patterns, five MspI patterns and two HinfI patterns. EcoRI showed no variation in the analysed sample. Additional patterns that did not account for known exon 2 DNA sequences were observed, suggesting the existence of novel ELA-DQB alleles. PCR-SSCP analysis exhibited seven different band patterns, and the number of bands per animal ranged from four to nine. Both methods indicated that at least two DQB genes are present. The presence of more than two alleles in each animal showed that the primers employed in this work are not specific for a unique DQB locus. The improvement of this PCR-RFLP method should provide a simple and rapid technique for an accurate definition of ELA-DQB typing in horses.

  5. Inhibitory effect of common microfluidic materials on PCR outcome

    KAUST Repository

    Kodzius, Rimantas

    2012-02-20

    Microfluidic chips have a variety of applications in the biological sciences and medicine. In contrast with traditional experimental approaches, microfluidics entails lower sample and reagent consumption, allows faster reactions and enables efficient separation. Additionally microfluidics offers other advantages accruing from the fluids’ various distinct behaviors, such as energy dissipation, fluidic resistance, laminar flow, and surface tension. Biological molecules suspended in fluid and transported through microfluidics channels interact with the channel-wall material. This interaction is even stronger in high surface-area-to-volume ratio (SAVR) microfluidic channels. Adsorption and inhibition of biomolecules occur when these materials come in contact with biomolecular reaction components. Polymerase chain reaction (PCR) is a thermal cycling procedure for amplifying target DNA. The PCR compatibility of silicon, silicon dioxide (SiO2) and other surfaces have been studied; however the results are inconclusive. Usually for protein-surface interaction measurements, bulky and expensive equipment is used, such as Atomic Force Microscopy (AFM), Scanning or Transmission Electron Microscopy (SEM, TEM), spectrophotometric protein concentration measurement, Fourier transform infrared spectroscopy (FTIR) or X-Ray photoelectron spectroscopy (XPS). \\tThe PCR reaction components include the DNA template, primers, DNA polymerase (the main component), dNTPs, a buffer, divalent ions (MgCl2), and KCl. \\tWe designed a simple, relatively quick measurement that only requires a PCR cycler; thus it mimics actual conditions in PCR cycling. In our study, we evaluated the inhibitory affect of different materials on PCR, which is one of the most frequently used enzymatic reactions in microfluidics. PCR reaction optimization through choice of surface materials is of the upmost importance, as it enables and improves enzymatic reaction in microfluidics. Our assessment of the PCR

  6. Micromecanizado de materiales cerámicos mediante láser de femtosegundo

    Directory of Open Access Journals (Sweden)

    Moreno, P.

    2005-02-01

    Full Text Available In this work, we present the application of ultrashort and intense laser pulses (110 fs @ 1 kHz; up to 1.1 mJ/pulse to the micromachining of ceramics, specifically RubalitTM708S, a material based on alumina and widely used as a substrate in microelectronics. The mechanism for removing material is the so called direct ablation. It differs from thermal ablation of conventional lasers in the practically total absence of thermal effects which produces a remarkable increase of quality and precision of the machining. By means of an optical diffraction-based technique we find out the energy density threshold to work in the direct ablation regime. Adjusting the energy per pulse as well as the number of pulses, we are able to drill holes of the desired diameter and depth. In addition, processing is developed in air. We also demonstrate that high quality fs-laser micromachining is suitable for every ceramic, whatever the mechanical properties, with similar working parameters. In order to show this point, we have also processed sintered SiN, a material of wide-ranging interest in industry.

    En este trabajo presentamos la aplicación de pulsos láser ultracortos (110 fs @ 1 kHz; hasta 1.1 mJ/pulso al micromecanizado de materiales cerámicos, en concreto RubalitTM708S, un material compuesto principalmente de alúmina y empleado en la industria microelectrónica. El mecanismo de eliminación de material es la ablación directa, que difiere de la ablación térmica empleada por los láseres convencionales en la prácticamente total ausencia de efectos térmicos, lo que redunda en un aumento significativo de la precisión y calidad del mecanizado. Mediante técnicas basadas en la difracción óptica determinamos el umbral de energía necesario para que tenga lugar el proceso de ablación directa. Con ese dato y regulando la energía por pulso y el número de pulsos somos capaces de producir mecanizados del diámetro y profundidad deseados. Además, el

  7. Efecto de la adición de Ag en Bi-2212 texturado mediante laser

    Directory of Open Access Journals (Sweden)

    Mora, M.

    2005-08-01

    Full Text Available The addition of Ag into Bi-2212 compounds has demonstrated to be a suitable method to improve both, the thermal and mechanical properties as well as the electrical ones. The final properties have been found to be in strong dependence of Ag content and the processing technique. In the present work the influence of Ag addition on Bi-2212 bulk materials grown from the melt, using a laser floating zone melting technique, has been studied. Samples with different Ag contents (0 to 40 wt.% were prepared for this work. The Bi-2212 + x wt.% Ag powders have been prepared by a sol-gel method via nitrates to assure total cation solution, small particle size and good homogeneity in the mixture. Cylindrical precursors, fabricated from these powders, were used as feed in a LFZ melting installation to obtain textured Bi-2212/Ag composites. The effect of the Ag addition on the microstructure is analysed as a function of Ag content. The changes on the microstructure are also correlated with the mechanical and superconducting properties.

    La incorporación de Ag en los compuestos de Bi-2212 ha demostrado ser un método adecuado para mejorar tanto las propiedades mecánicas, térmicas como eléctricas de estos materiales. Las propiedades finales dependen fuertemente de la cantidad de Ag añadida al sistema pero también del tipo de procesado que sufre. En el presente trabajo se realiza un estudio del efecto de la adición de Ag en materiales masivos Bi-2212 texturados mediante fusión zonal inducida por láser, con el objetivo de comprender el efecto de la adición de Ag en sistemas Bi-2212 que pasan totalmente por un fundido. Para ello se preparon muestras con diferentes contenidos en Ag (hasta el 40% en peso. Debido a la inmiscibilidad en estado sólido de la Ag y del Bi-2212, se ha utilizado un método de síntesis de estos materiales por medio de técnicas sol-gel para asegurar una buena homogeneidad y un tamaño de partícula reducido en la cerámica de

  8. Cleaning of contaminated soils with hydrocarbons by biocell; Saneamiento de suelos contaminados con hidrocarburos mediante biopilas

    Energy Technology Data Exchange (ETDEWEB)

    Iturbe-Arguelles, R.; Flores-Torres, C; Chavez-Lopez, C.; Roldan-Martin, A [Universidad Nacional Autonoma de Mexico, Mexico, D.F. (Mexico)

    2002-03-01

    In 1990 the Instituto de Ingenieria de la UNAM, initiated an evaluation through the soil and groundwater sampling and a risk health assessment in a Mexican refinery. An extended area was found contaminated with hydrocarbons. This area requires a soil remediation, taking into account that some zones present more than 30 000 mg/kg of Total Petroleum Hydrocarbons (TPH). Biopile system was recommended as the best remediation method to diminish TPG and some poliaromathic hydrocarbons (PAH). Therefore an experimental biopile of 30 m3 was constructed with contaminated soil. After 22 weeks, results show more than 80 % of TPH and PAH remotion. [Spanish] El grupo de saneamiento de suelos y acuiferos del Instituto de Ingenieria de la UNAM, inicio en 1999 la evaluacion de la contaminacion del subsuelo de una refineria en una zona costera del pais, mediante el muestreo de 425 puntos a 1.5 m de profundidad y con el analisis de los siguientes parametros: hidrocarburos totales del petroleo (HTP), hidrocarburo poliaromaticos (HAP), diesel, gasolina, metilterbutileter (MTBE) y los metales hierro, vanadio, zinc, cadmio, cromo y plomo. Asimismo, se lleva a cabo una evaluacion de riesgo a la salud a fin de determinar los niveles de limpieza de las areas contaminadas. Una vez realizado el estudio se propuso probar a nivel piloto dos tecnicas de saneamiento para las areas contaminadas con valores superiores a 30 000 mg/Kg de http, o bien, para las zonas en donde la evaluacion de riesgo a la salud indica la existencia de riesgo para uno o mas compuestos. Las tecnicas propuesta son biopilas y lavado de suelos con surfactantes. En este trabajo se presenta la prueba piloto con biopilas, de la cual se obtuvo una eficiencia de remocion de http del 80 porciento con cinco meses de operacion. Se muestra las partes de una biopila y se dan los resultados de la biopila experimental en la refineria Francisco I. Madero.

  9. ESTIMACIÓN ROBUSTA DE MODELOS ADITIVOS MEDIANTE EL ALGORITMO DE BACKFITTING

    Directory of Open Access Journals (Sweden)

    Luis P. Yapu Quispe

    2012-07-01

    Full Text Available En este trabajo se presenta un método de estimación y simulación de un modelo aditivo a dos variables mediante splines robustos, el método general puede ser aplicado con varias variables. El software utilizado para las simulaciones es S+ y se utiliza explícitamente la función smooth.splineRob en una implementación del algoritmo de backfitting. La función smooth.splineRob ha sido escrita en base al trabajo de Cantoni y Ronchetti [3], en el cual se pone énfasis en la selección robusta del parámetro de suavizamiento utilizando una versión robusta del Cp de Mallows, RCp, y de la validación cruzada, RCV. La existencia de datos extremos o no-normales en la parte estocástica de un modelo aditivo puede provocar una mala estimación del parámetro de suavizamiento, lo que tendrá influencia global en la estimación por splines. Para la etapa de simulación se realizan las estimaciones por splines clásicos y robustos (con estimación robusta del parámetro. La estimación obtenida es muy convincente pero el tiempo de ejecución del programa es relativamente elevado tanto para RCp y RCV, aun cuando, en ciertos casos, con pocas iteraciones robustas se obtienen ya resultados más útiles que la estimación clásica.

  10. Análisis del rendimiento academico mediante un modelo logit

    Directory of Open Access Journals (Sweden)

    María del Carmen Ibarra

    2010-07-01

    Full Text Available Este trabajo analiza el rendimiento académico de los estudiantes de la Facultad de Ingeniería de la Universidad Nacional de Misiones; la población objetivo está conformada por los alumnos de las cohortes 1999 a 2003 (589 estudiantes. Se define al rendimiento académico como el promedio de materias aprobadas anualmente y mediante la técnica estadística multivariada de Regresión Logística, se determina la incidencia que tienen diferentes factores de índole personal, socioeconómica y académica. Los resultados obtenidos permiten concluir que las variables significativas del rendimiento académico son: el promedio de calificaciones del nivel medio, el tipo de Institución donde cursó estos estudios y el número de asignaturas aprobadas en el primer año de carrera, siendo este último factor el más relevante, destacando la importancia de esta primera etapa de la carrera en los posteriores resultados académicos del estudiante. The purpose of this work is to analyse the determining factors which influence students’ performance at university. The research has been carried out on five (5 engineering students’ cohorts (1999-2003 from Universidad Nacional de Misiones (UNaM and includes 589 students. The academic performance is defined as average subjects approved annually. By means of the Logistic Regression technique, we determine the impact of different personal, socioeconomic and academic factors. The main conclusion that can be drawn is that students’ performance, is related to the grade point average (GPA at high school, the kind of high school (public or private students had attended, and the number of passing subjects in their first year at university. The latter being the most important factor, emphasizing the importance of this first stage at the Universtity in the student’s academic performance.

  11. Método para el Modelado y Prueba de Diagramas de Actividades Mediante Redes de Petri

    Directory of Open Access Journals (Sweden)

    Lionel Baquero Hernández

    2015-11-01

    Full Text Available Las redes de Petri son ideales para describir y estudiar sistemas que procesan información y con características concurrentes, asíncronas, distribuidas, paralelas, no determinísticas y/o estocásticas. . Sin embargo, la teoría clásica de estas redes se enfrenta al desafío de adecuarse a las nuevas necesidades de los sistemas informáticos que los ingenieros diseñan en la actualidad. Un diagrama de actividades representa una secuencia de actividades. Podría aplicarse a cualquier propósito, pero se considera especialmente útil para visualizar los flujos de trabajo y los procesos del negocio, o casos de uso. El presente trabajo se ha desarrollado con el objetivo de diseñar un método para el modelado y prueba de diagramas de actividades mediante redes de Petri. El método propuesto brinda una serie de pasos necesarios para modelar un diagrama de actividades con redes de Petri y a partir de dicha red obtener una representación matemática de la misma. Esta representación matemática se utiliza para con ella realizar una serie de operaciones basadas en el marcaje de la red que permiten realizar pruebas al diagrama. Este método es capaz de identificar errores comunes que se cometen en el modelado de los diagramas de actividades, lo cual se demostró a través del método de experimentación.

  12. Reconstrucción de defectos amplios en tronco mediante colgajo de perforante en piedra clave

    Directory of Open Access Journals (Sweden)

    J. Aguilera-Sáez

    Full Text Available El colgajo en piedra clave (Keystone Perforator Island Flap KPIF, es un colgajo local fasciocutáneo en isla que no requiere identificación de perforantes, descrito por Behan en 2003, que permite el cierre en un único acto quirúrgico tanto del defecto como de la zona dadora sin precisar, generalmente, autoinjerto cutáneo. Nuestro objetivo es el mostrar la utilidad de este colgajo para la cobertura de defectos amplios en tronco. De los más de 120 casos realizados en nuestro Servicio con esta técnica sin tener en cuenta la localización del problema entre abril de 2011 y abril de 2013, presentamos 5 casos que, tras resección quirúrgica por diferentes etiologías, fueron los que presentaron defectos de cobertura de mayor tamaño en tronco (entre 10 y 25 cm de diámetro máximo que fueron solventados satisfactoriamente mediante el KPIF. En todos obtuvimos un cierre directo tanto de la zona del defecto como de la zona dadora, sin complicaciones durante el acto quirúrgico ni en el postoperatorio, con resultados estéticos óptimos, sin deformidades significativas ni alteraciones funcionales. Creemos que el KPIF es un colgajo óptimo para la cobertura de defectos amplios en el tronco ya que permite cubrir el defecto y hacer un cierre primario de la zona dadora generando poca morbilidad, con un buen resultado estético, con un corto tiempo quirúrgico y con un bajo coste.

  13. PoV-GAME: PUNTOS DE VISTA MEDIANTE JUEGOS PoV-GAME: VIEWPOINTS THROUGH GAMES

    Directory of Open Access Journals (Sweden)

    Carlos Zapata-Jaramillo

    2012-06-01

    Full Text Available El desarrollo de proyectos de software considera importante la consistencia entre la información entregada por todos los interesados de una aplicación. En otras palabras, se procura llegar a acuerdos entre los diferentes puntos de vista de cada actor y llevarlos a cabo durante todo el proyecto. Con los métodos de enseñanza tradicionales se procura formar habilidades en este campo, pero no se afianzan mediante la práctica que, generalmente, se emplea para el aprendizaje en este campo. Por ello, en este artículo se propone ''PoV-GAME'', una nueva forma de ''jugar aprendiendo'', la cual busca fortalecer algunos conceptos básicos en la ingeniería de software como la consistencia en la información que se maneja a lo largo de un proyecto y afianzar la importancia de los puntos de vista en el desarrollo de un producto de software. Posteriormente, se analizan los resultados obtenidos luego de realizar el juego en diferentes grupos de estudiantes.Information for software applications is delivered by the stakeholders. Software development projects need consistency as an important aspect of such information. In other words, agreement among the different viewpoints must be reached in order to be implemented in the project. Well-known teaching methods try to create capabilities in this field, but they fail in reinforcing them in practice, which is commonly used for teaching in this field. Therefore, in this paper we propose ''PoV GAME'' a new way of ''play-to-learn''. We aim the reinforcement of some basic concepts about software engineering such as consistency in the information handled throughout the project, and the importance of viewpoints in the software product development. Subsequently, we analyze the results obtained after holding the game in several groups of students.

  14. Diagnóstico de Fallos en Sistemas Híbridos Mediante Anidamiento Latente de Fallos

    Directory of Open Access Journals (Sweden)

    Orestes Llanes Santiago

    2010-09-01

    Full Text Available Normal 0 21 false false false ES-TRAD X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabla normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} Este artículo presenta un nuevo enfoque para la detección y diagnóstico de fallos (SDDF en sistemas híbridos denominado de Anidamiento Latente de Fallos. La misma está basada en el uso de Redes de Petri Coloreadas (RdPCs que aportan su capacidad intrínseca de síntesis y de implementación en modelos realizables. Esta técnica se contrapone al clásico fenómeno de explosión combinacional que se produce mediante la utilización de metodologías basadas en Máquinas de Estados Finitos (MEFs.

  15. Caracterización microestructural de aleaciones base cobre obtenidas mediante molienda reactiva

    Directory of Open Access Journals (Sweden)

    Palma, R.

    2010-06-01

    Full Text Available The micro and nanostructure of Cu-Al, Cu-V and Cu-Ti alloys produced by reactive milling were analyzed using X-ray diffraction (XRD and transmission electron microscopy (TEM. Samples with different milling times (t= 0, 10, 20 and 30 h were considered. The grain size, dislocation density and residual microstrain were evaluated form the XRD data using the Williamson-Hall and Klug-Alexander methods. The evolution of texture as a function of milling time was also studied using XRD. It was found, using TEM, that the grain size and dispersoid size were nanometric in all three alloys considered.

    Se analizó la micro y nano estructura de aleaciones Cu-Al, Cu-V y Cu-Ti obtenidas por molienda reactiva, mediante difracción de rayos X (XRD y microscopía electrónica de transmisión (TEM. Se consideraron muestras con distintos tiempos de molienda (t= 0, 10, 20 y 30 h. A partir de los datos XRD, usando los métodos de Williamson- Hall y Klug-Alexander, se evaluaron el tamaño de grano, la densidad de dislocaciones y la microdeformación residual; también se estudió la evolución de la textura de la matriz de cobre en función del tiempo de molienda. En los polvos molidos durante 30 h, de las tres aleaciones consideradas, se encontró, por TEM, que los tamaños de grano y de los dispersoides desarrollados son nanométricos.

  16. Tratamiento quirúrgico de las lesiones intradurales extramedulares mediante hemilaminectomía

    Science.gov (United States)

    Villalonga, Juan F.; Cervio, Andrés

    2017-01-01

    Resumen Objetivo: Evaluar la utilidad de la hemilaminectomía como abordaje quirúrgico en pacientes con tumores intradurales-extramedulares. Material y métodos: Estudio descriptivo retrospectivo que incluye a 53 pacientes en los que se utilizó la hemilaminectomía como abordaje a tumores intradurales-extramedulares durante el periodo junio de 2006 a diciembre de 2015. Se analizaron datos demográficos, signo-sintomatología preoperatoria, características imagenológicas, hallazgos intraoperatorios, estirpe histológico y complicaciones post-quirúrgicas. El periodo de seguimiento promedio fue de 48.9 meses (6-120 meses). Resultados: Cincuenta y tres pacientes con tumores intradurales-extramedulares fueron intervenidos mediante hemilaminectomía. La serie incluyó 5 tumores cervicales, 24 dorsales y 24 lumbares. El análisis histológico reveló 28 neurinomas, 11 meningiomas, 7 ependimomas y 7 “lesiones varias”. En el 96% de los casos se efectuó una exéresis total sin causar déficit neurológico agregado. No se evidenció recidiva en ninguno de los casos durante el periodo de seguimiento. Conclusión: La hemilaminectomía constituye una vía efectiva para la resección de tumores intradurales-extramedulares lateralizados a nivel cervicodorsal. Mientras que a nivel lumbar esta técnica puede ser también útil en lesiones de línea media. PMID:29142776

  17. PREACONDICIONAMIENTO DE Pinus engelmannii Carr. MEDIANTE DÉFICIT DE RIEGO EN VIVERO

    Directory of Open Access Journals (Sweden)

    Israel J. Ávila-Flores

    2014-01-01

    Full Text Available Con el objetivo de mejorar el preacondicionamiento de plántulas de Pinus engelmannii Carr. en vivero, se evaluaron tres frecuencias de riego: 48, 96 y 192 h. Las plántulas de ocho meses de edad se evaluaron durante 40 días. El ensayo se desarrolló en condiciones de invernadero en el vivero del Campo Experimental Valle del Guadiana del INIFAP. Los datos se analizaron mediante prueba de permutaciones. Los resultados muestran que el riego cada 48 h causó los mejores efectos en altura (7.70 cm, diámetro (4.38 mm, biomasa total (6.59 g e índice de calidad de Dickson (0.55. Por otro lado, el estrés hídrico originado por el tratamiento con riego cada 192 h (-1.22 MPa limitó el crecimiento de las plántulas; sin embargo, el índice de lignificación (26.63 fue significativa - mente mejor ( P = 0.0001 con esta frecuencia de riego. A pesar de haberse encontrado diferencias im - portantes ( P = 0.0001 en el potencial hídrico y en las variables morfológicas, no se encontró evidencia suficiente que permita definir un tratamiento absolutamente superior para mejorar el preacondiciona - miento de las plántulas en vivero. Por lo tanto, es recomendable evaluar dichos tratamientos durante mayor tiempo, además de evaluarlos en campo.

  18. Combate del moho gris (Botrytis cinerea de la fresa mediante Gliocladium roseum

    Directory of Open Access Journals (Sweden)

    Néstor Chaves

    2004-01-01

    Full Text Available En la zona de Poasito de Alajuela, se evaluó la acción del antagonista Gliocladium roseum, en forma individual y en conjunto con los fungicidas empleados en la finca, para el combate de Botrytis cinerea en fresa; comparándose los resultados contra los obtenidos con el manejo comercial. Se empleó un diseño de bloques completos al azar con 4 repeticiones y se hicieron aplicaciones semanales del antagonista (a una concentración ³ 107 conidios ml-1 durante un período aproximado de 4 meses (julio-octubre del 2000. Se evaluó la incidencia de moho gris en condiciones de campo y poscosecha, así como el efecto de los fungicidas aplicados sobre la germinación de los conidios del antagonista, mediante una prueba in vitro. Se obtuvo un combate más efectivo de la enfermedad en condiciones de campo al emplear el biocontrolador sólo o en conjunto con los fungicidas, con respecto al manejo comercial que se hace de la misma. En poscosecha, el desempeño del antagonista fue estadísticamente igual al del combate químico. Estos resultados muestran que los fungicidas aplicados no afectan considerablemente al antagonista, lo que se corroboró con la prueba in vitro. Al emplear G. roseum para el combate de B. cinerea no sólo se logra combatir efectivamente a este, sino también el resto de los patógenos (Colletotrichum, Phytophthora, Rhizoctonia, Rhizopus,Alternaria, Fusarium, Verticillium y Penicillium, ya que el porcentaje de frutas sanas es mayor al integrar la acción del antagonista al manejo de enfermedades de la finca. Sin embargo, estas diferencias no son estadísticamente significativas. Por lo anterior se concluye que G. roseum constituye una posible alternativa de manejo integrado del moho gris en fresa.

  19. Estudio del crecimiento de Prioria copaifera (Caesalpinaceae mediante técnicas dendrocronológicas

    Directory of Open Access Journals (Sweden)

    Jorge Andrés Giraldo Jiménez

    2011-12-01

    Full Text Available El cativo (Prioria copaifera G. forma bosques dominados por la especie llamados cativales. Durante más de 70 años el cativo ha sido la base principal de la industria maderera en el Darién colombiano, siendo, por su alta productividad y dominancia uno de los bosques tropicales más fáciles de ordenar sosteniblemente. El propósito de esta investigación es modelar el crecimiento del diámetro y el volumen del cativo en función de la edad así como las tasas instantáneas, medias y relativas, empleando anillos de crecimiento. La anualidad de los anillos se demostró usando el efecto de las bombas nucleares y mediante cofechado. Con base en los anillos de crecimiento radiales se modelo el crecimiento del diámetro y del volumen. Esta especie alcanza 40cm de diámetro en 90 años y presenta crecimiento medio de 0.31cm/año. El lapso vital de la especie es de aproximadamente 614 años. El incremento corriente anual máximo del volumen a los 90 años es 0.033m3/año, e iguala al incremento medio a los 145 años con 0.018m3/año. La tasa media de crecimiento absoluta es 0.021m3/año. Los resultados aquí obtenidos sirven como base técnica para el manejo sostenible de los cativales.

  20. Droplet digital PCR (ddPCR) vs quantitative real-time PCR (qPCR) approach for detection and quantification of Merkel cell polyomavirus (MCPyV) DNA in formalin fixed paraffin embedded (FFPE) cutaneous biopsies.

    Science.gov (United States)

    Arvia, Rosaria; Sollai, Mauro; Pierucci, Federica; Urso, Carmelo; Massi, Daniela; Zakrzewska, Krystyna

    2017-08-01

    Merkel cell polyomavirus (MCPyV) is associated with Merkel cell carcinoma and high viral load in the skin was proposed as a risk factor for the occurrence of this tumour. MCPyV DNA was detected, with lower frequency, in different skin cancers but since the viral load was usually low, the real prevalence of viral DNA could be underestimated. To evaluate the performance of two assays (qPCR and ddPCR) for MCPyV detection and quantification in formalin fixed paraffin embedded (FFPE) tissue samples. Both assays were designed to simultaneous detection and quantification of both MCPyV as well as house-keeping DNA in clinical samples. The performance of MCPyV quantification was investigated using serial dilutions of cloned target DNA. We also evaluated the applicability of both tests for the analysis of 76 FFPE cutaneous biopsies. The two approaches resulted equivalent with regard to the reproducibility and repeatability and showed a high degree of linearity in the dynamic range tested in the present study. Moreover, qPCR was able to quantify ≥10 5 copies per reaction, while the upper limit of ddPCR was 10 4 copies. There was not significant difference between viral load measured by the two methods The detection limit of both tests was 0,15 copies per reaction, however, the number of positive samples obtained by ddPCR was higher than that obtained by qPCR (45% and 37% respectively). The ddPCR represents a better method for detection of MCPyV in FFPE biopsies, mostly these containing low copies number of viral genome. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Species identification of Cannabis sativa using real-time quantitative PCR (qPCR).

    Science.gov (United States)

    Johnson, Christopher E; Premasuthan, Amritha; Satkoski Trask, Jessica; Kanthaswamy, Sree

    2013-03-01

    Most narcotics-related cases in the United States involve Cannabis sativa. Material is typically identified based on the cystolithic hairs on the leaves and with chemical tests to identify of the presence of cannabinoids. Suspect seeds are germinated into a viable plant so that morphological and chemical tests can be conducted. Seed germination, however, causes undue analytical delays. DNA analyses that involve the chloroplast and nuclear genomes have been developed for identification of C. sativa materials, but they require several nanograms of template DNA. Using the trnL 3' exon-trnF intragenic spacer regions within the C. sativa chloroplast, we have developed a real-time quantitative PCR assay that is capable of identifying picogram amounts of chloroplast DNA for species determination of suspected C. sativa material. This assay provides forensic science laboratories with a quick and reliable method to identify an unknown sample as C. sativa. © 2013 American Academy of Forensic Sciences.

  2. Instalación eléctrica de una vivienda unifamiliar aislada mediante suministro de energías renovables

    OpenAIRE

    LOZANO VALLADOLID, FERNANDO

    2015-01-01

    [ES] Instalación eléctrica con grado de electrificación elevada de una vivienda unifamiliar aislada mediante suministro de energías renovables (solar, eólica, geotérmicas). Lozano Valladolid, F. (2015). Instalación eléctrica de una vivienda unifamiliar aislada mediante suministro de energías renovables. http://hdl.handle.net/10251/58751. TFGM

  3. Valoración mediante una encuesta de la negativa a la vacunación frente al virus del papiloma humano: estudio de los motivos para no vacunar

    OpenAIRE

    Alonso Benito, Cristina

    2017-01-01

    Treball de Final de Grau en Medicina. Codi: MD1158. Curs acadèmic 2016-2017 Objetivo: La vacuna frente al Virus del Papiloma Humano (VPH) ha demostrado ser la estrategia más eficaz en la prevención del cáncer de cérvix. Desde el año 2008 forma parte del Calendario de Vacunación Sistemática Infantil en la Comunidad Valenciana. Sin embargo, se ha observado que la tasa de vacunación no alcanza los objetivos recomendados. Este trabajo tiene como fin, identificar los motivos por los...

  4. Design and optimization of reverse-transcription quantitative PCR experiments.

    Science.gov (United States)

    Tichopad, Ales; Kitchen, Rob; Riedmaier, Irmgard; Becker, Christiane; Ståhlberg, Anders; Kubista, Mikael

    2009-10-01

    Quantitative PCR (qPCR) is a valuable technique for accurately and reliably profiling and quantifying gene expression. Typically, samples obtained from the organism of study have to be processed via several preparative steps before qPCR. We estimated the errors of sample withdrawal and extraction, reverse transcription (RT), and qPCR that are introduced into measurements of mRNA concentrations. We performed hierarchically arranged experiments with 3 animals, 3 samples, 3 RT reactions, and 3 qPCRs and quantified the expression of several genes in solid tissue, blood, cell culture, and single cells. A nested ANOVA design was used to model the experiments, and relative and absolute errors were calculated with this model for each processing level in the hierarchical design. We found that intersubject differences became easily confounded by sample heterogeneity for single cells and solid tissue. In cell cultures and blood, the noise from the RT and qPCR steps contributed substantially to the overall error because the sampling noise was less pronounced. We recommend the use of sample replicates preferentially to any other replicates when working with solid tissue, cell cultures, and single cells, and we recommend the use of RT replicates when working with blood. We show how an optimal sampling plan can be calculated for a limited budget. .

  5. Droplet digital PCR technology promises new applications and research areas.

    Science.gov (United States)

    Manoj, P

    2016-01-01

    Digital Polymerase Chain Reaction (dPCR) is used to quantify nucleic acids and its applications are in the detection and precise quantification of low-level pathogens, rare genetic sequences, quantification of copy number variants, rare mutations and in relative gene expressions. Here the PCR is performed in large number of reaction chambers or partitions and the reaction is carried out in each partition individually. This separation allows a more reliable collection and sensitive measurement of nucleic acid. Results are calculated by counting amplified target sequence (positive droplets) and the number of partitions in which there is no amplification (negative droplets). The mean number of target sequences was calculated by Poisson Algorithm. Poisson correction compensates the presence of more than one copy of target gene in any droplets. The method provides information with accuracy and precision which is highly reproducible and less susceptible to inhibitors than qPCR. It has been demonstrated in studying variations in gene sequences, such as copy number variants and point mutations, distinguishing differences between expression of nearly identical alleles, assessment of clinically relevant genetic variations and it is routinely used for clonal amplification of samples for NGS methods. dPCR enables more reliable predictors of tumor status and patient prognosis by absolute quantitation using reference normalizations. Rare mitochondrial DNA deletions associated with a range of diseases and disorders as well as aging can be accurately detected with droplet digital PCR.

  6. Result Variation and Efficiency Kinetics in Real-Time PCR

    Directory of Open Access Journals (Sweden)

    Reza Shahsiah

    2010-10-01

    Full Text Available Fluorescent monitoring of DNA amplification is the basis of real-time PCR. Absolute quantification can be achieved using a standard curve method. The standard curve is constructed by amplifying known amounts of standards under identical conditions to that of the samples.The objective of the current study is to propose a mathematical model to assess the acceptability of PCR resulys.Four commercial standards for HCV-RNA (hepatitis C virus RNA along with 6 patient samples were measured by real-time PCR, using two different RT-PCR reagents. The standard deviation of regression (Sy,x was calculated for each group of standard and compared by F-Test. The efficiency kinetics was computed by logistic regression, c2 goodness of fit test was preformed to assess the appropriateness of the efficiency curves.Calculated efficiencies were not significantly different from the value predicted by logistic regression model. Reactions with more variation showed less stable efficiency curves, with wider range of amplification efficiencies.Amplification efficiency kinetics can be computed by fitting a logistic regression curve to the gathered fluorescent data of each reaction. This model can be employed to assess the acceptability of PCR results calculated by standard curve method.

  7. Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR (PMA-qPCR).

    Science.gov (United States)

    Li, Dan; Tong, Tiezheng; Zeng, Siyu; Lin, Yiwen; Wu, Shuxu; He, Miao

    2014-02-01

    The detection of viable bacteria in wastewater treatment plants (WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide (PMA) combined with the quantitative polymerase chain reaction (PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 micromol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.

  8. PCR detection of Bartonella spp. in the dog

    Directory of Open Access Journals (Sweden)

    Jarmila Konvalinová

    2014-01-01

    Full Text Available Our study aimed at using PCR to identify the incidence of Bartonella spp. in blood of dogs. Altogether 286 dogs of 92 breeds aged 3 month to 17 years were tested from October 2008 to December 2009. Healthy dogs as well as dogs with various clinical symptoms of disease were included in the group. Samples were tested by polymerase chain reaction (PCR specific for the presence of Bartonella spp. Following the DNA examination in 286 dogs by PCR and subsequent sequencing, two samples were identified as Bartonella henselae (0.7%. Other species of Bartonella were not found. It was the first time in the Czech Republic when incidence of Bartonella spp. was determined in dogs.

  9. [Application of rapid PCR to authenticate medicinal snakes].

    Science.gov (United States)

    Chen, Kang; Jiang, Chao; Yuan, Yuan; Huang, Lu-Qi; Li, Man

    2014-10-01

    To obtained an accurate, rapid and efficient method for authenticate medicinal snakes listed in Chinese Pharmacopoeia (Zaocysd humnades, Bungarus multicinctus, Agkistrodon acutus), a rapid PCR method for authenticate snakes and its adulterants was established based on the classic molecular authentication methods. DNA was extracted by alkaline lysis and the specific primers were amplified by two-steps PCR amplification method. The denatured and annealing temperature and cycle numbers were optimized. When 100 x SYBR Green I was added in the PCR product, strong green fluorescence was visualized under 365 nm UV whereas adulterants without. The whole process can complete in 30-45 minutes. The established method provides the technical support for authentication of the snakes on field.

  10. RUCS: Rapid identification of PCR primers for unique core sequences

    DEFF Research Database (Denmark)

    Thomsen, Martin Christen Frølund; Hasman, Henrik; Westh, Henrik

    2017-01-01

    Designing PCR primers to target a specific selection of whole genome sequenced strains can be a long, arduous, and sometimes impractical task. Such tasks would benefit greatly from an automated tool to both identify unique targets, and to validate the vast number of potential primer pairs...... for the targets in silico . Here we present RUCS, a program that will find PCR primer pairs and probes for the unique core sequences of a positive genome dataset complement to a negative genome dataset. The resulting primer pairs and probes are in addition to simple selection also validated through a complex...... in silico PCR simulation. We compared our method, which identifies the unique core sequences, against an existing tool called ssGeneFinder, and found that our method was 6.5-20 times more sensitive. We used RUCS to design primer pairs that would target a set of genomes known to contain the mcr-1 colistin...

  11. Analysis of Multiallelic CNVs by Emulsion Haplotype Fusion PCR.

    Science.gov (United States)

    Tyson, Jess; Armour, John A L

    2017-01-01

    Emulsion-fusion PCR recovers long-range sequence information by combining products in cis from individual genomic DNA molecules. Emulsion droplets act as very numerous small reaction chambers in which different PCR products from a single genomic DNA molecule are condensed into short joint products, to unite sequences in cis from widely separated genomic sites. These products can therefore provide information about the arrangement of sequences and variants at a larger scale than established long-read sequencing methods. The method has been useful in defining the phase of variants in haplotypes, the typing of inversions, and determining the configuration of sequence variants in multiallelic CNVs. In this description we outline the rationale for the application of emulsion-fusion PCR methods to the analysis of multiallelic CNVs, and give practical details for our own implementation of the method in that context.

  12. Typing DNA profiles from previously enhanced fingerprints using direct PCR.

    Science.gov (United States)

    Templeton, Jennifer E L; Taylor, Duncan; Handt, Oliva; Linacre, Adrian

    2017-07-01

    Fingermarks are a source of human identification both through the ridge patterns and DNA profiling. Typing nuclear STR DNA markers from previously enhanced fingermarks provides an alternative method of utilising the limited fingermark deposit that can be left behind during a criminal act. Dusting with fingerprint powders is a standard method used in classical fingermark enhancement and can affect DNA data. The ability to generate informative DNA profiles from powdered fingerprints using direct PCR swabs was investigated. Direct PCR was used as the opportunity to generate usable DNA profiles after performing any of the standard DNA extraction processes is minimal. Omitting the extraction step will, for many samples, be the key to success if there is limited sample DNA. DNA profiles were generated by direct PCR from 160 fingermarks after treatment with one of the following dactyloscopic fingerprint powders: white hadonite; silver aluminium; HiFi Volcano silk black; or black magnetic fingerprint powder. This was achieved by a combination of an optimised double-swabbing technique and swab media, omission of the extraction step to minimise loss of critical low-template DNA, and additional AmpliTaq Gold ® DNA polymerase to boost the PCR. Ninety eight out of 160 samples (61%) were considered 'up-loadable' to the Australian National Criminal Investigation DNA Database (NCIDD). The method described required a minimum of working steps, equipment and reagents, and was completed within 4h. Direct PCR allows the generation of DNA profiles from enhanced prints without the need to increase PCR cycle numbers beyond manufacturer's recommendations. Particular emphasis was placed on preventing contamination by applying strict protocols and avoiding the use of previously used fingerprint brushes. Based on this extensive survey, the data provided indicate minimal effects of any of these four powders on the chance of obtaining DNA profiles from enhanced fingermarks. Copyright © 2017

  13. Soil Baiting, Rapid PCR Assay and Quantitative Real Time PCR to Diagnose Late Blight of Potato in Quarantine Programs

    Directory of Open Access Journals (Sweden)

    Touseef Hussain

    2018-05-01

    Full Text Available Phytophthora infestans (mont de Bary is a pathogen of great concern across the globe, and accurate detection is an important component in responding to the outbreaks of potential disease. Although the molecular diagnostic protocol used in regulatory programs has been evaluated but till date methods implying direct comparison has rarely used. In this study, a known area soil samples from potato fields where light blight appear every year (both A1 and A2 mating type was assayed by soil bait method, PCR assay detection and quantification of the inoculums. Suspected disease symptoms appeared on bait tubers were further confirmed by rapid PCR, inoculums were quantified through Real Time PCR, which confirms presence of P. infestans. These diagnostic methods can be highly correlated with one another. Potato tuber baiting increased the sensitivity of the assay compared with direct extraction of DNA from tuber and soil samples. Our study determines diagnostic sensitivity and specificity of the assays to determine the performance of each method. Overall, molecular techniques based on different types of PCR amplification and Real-time PCR can lead to high throughput, faster and more accurate detection method which can be used in quarantine programmes in potato industry and diagnostic laboratory.

  14. Droplet Digital™ PCR Next-Generation Sequencing Library QC Assay.

    Science.gov (United States)

    Heredia, Nicholas J

    2018-01-01

    Digital PCR is a valuable tool to quantify next-generation sequencing (NGS) libraries precisely and accurately. Accurately quantifying NGS libraries enable accurate loading of the libraries on to the sequencer and thus improve sequencing performance by reducing under and overloading error. Accurate quantification also benefits users by enabling uniform loading of indexed/barcoded libraries which in turn greatly improves sequencing uniformity of the indexed/barcoded samples. The advantages gained by employing the Droplet Digital PCR (ddPCR™) library QC assay includes the precise and accurate quantification in addition to size quality assessment, enabling users to QC their sequencing libraries with confidence.

  15. Single base pair mutation analysis by PNA directed PCR clamping

    DEFF Research Database (Denmark)

    Ørum, H.; Nielsen, P.E.; Egholm, M.

    1993-01-01

    A novel method that allows direct analysis of single base mutation by the polymerase chain reaction (PCR) is described. The method utilizes the finding that PNAs (peptide nucleic acids) recognize and bind to their complementary nucleic acid sequences with higher thermal stability and specificity...... allows selective amplification/suppression of target sequences that differ by only one base pair. Finally we show that PNAs can be designed in such a way that blockage can be accomplished when the PNA target sequence is located between the PCR primers....

  16. Identification of Meat Species by Polymerase Chain Reaction (PCR) Technique

    OpenAIRE

    İLHAK, O. İrfan; ARSLAN, Ali

    2014-01-01

    The origin of horse, dog, cat, bovine, sheep, porcine, and goat meat was determined by the polymerase chain reaction (PCR) technique, using species-specific primers. Test mixtures of meat were prepared by adding 5%, 2.5%, 1%, 0.5%, and 0.1% levels of pork, horse, cat, or dog meat to beef, sheep, and goat meat. Samples taken from those combinations were analyzed by PCR for species determination. Mitochondrial DNA (mt DNA) fragments of 439, 322, 274, 271, 225, 212, and 157 bp for horse, dog, ca...

  17. Typing of Y chromosome SNPs with multiplex PCR methods

    DEFF Research Database (Denmark)

    Sanchez Sanchez, Juan Jose; Børsting, Claus; Morling, Niels

    2005-01-01

    We describe a method for the simultaneous typing of Y-chromosome single nucleotide polymorphism (SNP) markers by means of multiplex polymerase chain reaction (PCR) strategies that allow the detection of 35 Y chromosome SNPs on 25 amplicons from 100 to 200 pg of chromosomal deoxyribonucleic acid...... factors for the creation of larger SNP typing PCR multiplexes include careful selection of primers for the primary amplification and the SBE reaction, use of DNA primers with homogenous composition, and balancing the primer concentrations for both the amplification and the SBE reactions....

  18. Detection of hepatitis C virus RNA using reverse transcription PCR

    International Nuclear Information System (INIS)

    Yap, S.F.

    1998-01-01

    Detection of the viral genome (HCV RNA) is by a combination of cDNA synthesis and PCR followed by gel analysis and/or hybridization assay. In principle, cDNA is synthesized using the viral RNA as template and the enzyme, reverse transcriptase. The cDNA is then amplified by PCR and the product detected. Agarose gel electrophoresis provides a rapid and simple detection method; however, it is non-quantitative. The assay protocol described in this paper is adapted from that published by Chan et al. Comments on various aspects of the assay are based on experience with the method in our laboratory

  19. Fecal specimens preparation methods for PCR diagnosis of human taeniosis

    Directory of Open Access Journals (Sweden)

    Nunes Cáris Maroni

    2006-01-01

    Full Text Available Sample preparation and DNA extraction protocols for DNA amplification by PCR, which can be applied in human fecal samples for taeniasis diagnosis, are described. DNA extracted from fecal specimens with phenol/chloroform/isoamilic alcohol and DNAzol® reagent had to be first purified to generate fragments of 170 pb and 600 pb by HDP2-PCR. This purification step was not necessary with the use of QIAmp DNA stool mini kit®. Best DNA extraction results were achieved after eggs disruption with glass beads, either with phenol/chloroform/isoamilic alcohol, DNAzol® reagent or QIAmp DNA stool mini kit®.

  20. Obtención de Si3N4 mediante SHS

    Directory of Open Access Journals (Sweden)

    Rodríguez, M. A.

    2003-04-01

    parámetros que definen la propia reacción. Con la información obtenida se propone el mecanismo predominante de la síntesis del Nitruro de Silicio mediante SHS.

  1. Efecto del argon en películas CNxHy depositadas mediante ECR-CVD

    Directory of Open Access Journals (Sweden)

    Albella, J. M.

    2004-04-01

    Full Text Available Carbon nitride films have been deposited by ECR-CVD, from Ar/CH4/N2 gas mixtures with different methane concentrations. Infrared Spectroscopy (IRS and Elastic Recoil Detection Analysis (ERDA have been used for films characterisation and Optical Emission Spectroscopy (OES for plasma analysis. Argon concentration in the gas mixture controls the growth rate as well as the composition of the film. In the proposed model, argon plays a key role in the activation of methane molecules. Also, during the growth of the film, two processes may be considered: i Film formation and ii Etching of the growing surface. Changing the gas mixture composition affects both processes, which results in films with different composition and structure as well as different deposition rates.Se ha estudiado el efecto del argon durante el proceso de CVD asistido por un plasma ECR para la síntesis de películas de nitruro de carbono (CNxHy a partir de mezclas gaseosas Ar/CH4/N2 con diferente contenido de metano. Las películas depositadas han sido analizadas mediante espectroscopía infrarroja (IRS y ERDA (Elastic Recoil Detection Analysis, y el análisis del plasma ha sido realizado utilizando la técnica de espectroscopía de emisión óptica (OES. La velocidad de deposición y la composición de las películas depositadas se encuentran determinadas por la concentración de argon en la mezcla gaseosa. Se propone un modelo, según el cual el argon juega un papel fundamental como activador de las moléculas de metano. El modelo propuesto incluye dos procesos simultáneos durante el crecimiento de las capas : i formación de la capa y ii ataque de la superficie de crecimiento. Según la composición de la mezcla gaseosa se favorece uno u otro proceso, lo que conduce a velocidades de deposición diferentes así como a depósitos con diferente composición y estructura atómica.

  2. Standardizzazione isogravitá di un case-mix ospedaliero mediante Charlson index

    Directory of Open Access Journals (Sweden)

    G. Messina

    2003-05-01

    Full Text Available

    Introduzione: la comorbidità è un importante fattore confondente negli studi epidemiologici valutativi dell’assistenza ospedaliera. Diversi strumenti di risk adjustment misurano la complessità della malattia, consentendo di correlarla al consumo di risorse assistenziali, agli esiti, nonchè di confrontare studi eseguiti in tempi e realtà diversi.

    Obiettivi: - Standardizzare per complessità casistica la mortalità dei pazienti assistiti da un grande ospedale; - Identificare le variabili in grado di migliorare la capacità predittiva di mortalità intraospedaliera (IM. Materiali e

    Metodi: sono state analizzate 40.801 schede di dimissione prodotte dal Policlinico Senese nel 2001. Sono stati studiati i tassi di IM specifici per Charlson Index Score (CSI: quest’ultimo considera 19 categorie di patologia e si basa sull’ ICD-IX-CM. Le variabili studiate mediante analisi bivariate e regressione logistica, sono state: CSI (codificato in 5 livelli 0, 1, 2, 3, 4, lunghezza del ricovero (LR, sesso ed età. Risultati: CSI è risultato associato con IM (p‹0.001. Sesso, età e LR sono risultati associati sia con CSI (p‹0.001 che IM (p‹0.001. Confrontando i 4 livelli di CSI con quello di riferimento (0, le seguenti Odds Ratio (OR di IM sono state trovate: Livello 1 verso livello 0 OR: 6.79 (p‹0.001, Livello 2 verso livello 0 OR: 15.8 (p‹0.001, Livello 3 verso livello 0 OR: 9.36 (p‹0.001, Livello 4 verso livello 0 OR: 7.4 (p‹0.001. La variabile sesso non è risultata aver un effetto confondente tra CSI e IM al contrario delle variabili LR ed età.

    Conclusioni: il CSI aiuta a valutare (predire il rischio di mortalità intraospedaliera, sebbene in modo non lineare. Abbiamo sempre rilevato valori più alti di mortalità confrontando i livelli 1, 2, 3, e 4 con quello di riferimento (0. In particolare il valore più alto lo ha raggiunto il secondo livello

  3. EDUCACIÓN MEDIANTE ENCICLOPEDIAS VISUALES TEMÁTICAS: INTYPEDIA UN CASO DE ÉXITO

    Directory of Open Access Journals (Sweden)

    ALFONSO MUÑOZ MUÑOZ

    2013-01-01

    Full Text Available La presente investigación recoge una experiencia de innovación educativa que demuestra la utilidad de las enciclopedias visuales temáticas para la formación académica a través de Internet. Los objetivos de esta experiencia recaen en verificar si es posible crear enciclopedias visuales temáticas que mediante recursos audiovisuales permitan el aprendizaje de aspectos complejos de una manera asequible. Con el interés de obtener datos que permitan comprobar la idoneidad de este tipo de recursos formativos, se construye una enciclopedia visual temática centrada en aspectos de la seguridad de la información y protección de las comunicaciones digitales. Para el diseño de esta enciclopedia se considera una metodología de diseño basada en la definición de una temática de interés, tipo de aprendizaje, formato y calidad de las unidades temáticas, difusión y coste, etc. Esta enciclopedia visual, el proyecto Intypedia, novedoso a escala mundial, está compuesta por lecciones en vídeo de unos 12 minutos y material docente complementario. El lenguaje utilizado permite el correcto seguimiento de las lecciones tanto a expertos en seguridad como internautas en general. Por tanto, la calidad y claridad de estos contenidos se convierten en un valor añadido del proyecto. Los resultados derivados reflejan la utilidad de Intypedia. En 2 años de vida la enciclopedia recibe miles de visitas al mes y se ha convertido en una enciclopedia de referencia en Iberoamérica y en las redes sociales en las que está presente. Algunos de los resultados más importantes son: 232.786 reproducciones únicas de las lecciones, siendo los consumidores principalmente internautas, profesionales y técnicos de España, Iberoamérica y EE.UU.; más de 176.000 documentos descargados, en torno a los 2.100 seguidores uniendo las redes sociales y una visibilidad en Google con más de 40.000 entradas que enlazan al proyecto.

  4. Identificación de territorios críticos en salud materna mediante indicadores

    Directory of Open Access Journals (Sweden)

    Marcos Paz Ballivián

    2002-07-01

    Full Text Available Objetivos. El Ministerio de Salud y Previsión Social de Bolivia convocó a principios de 2000 a varios expertos para que, a partir de la información disponible, elaboraran un método que permitiera identificar territorios críticos en un tema de por sí prioritario --la salud materna y neonatal-- y crear un mapa de la situación y de la capacidad instalada en las 112 provincias nacionales. En este estudio se describe el método utilizado. Métodos. Se crearon índices de la situación de salud y de la capacidad instalada. Los pasos seguidos en este proceso fueron los siguientes: 1 identificación de las variables incluidas en cada índice; 2 ponderación de las variables de cada índice; creación de una fórmula para cada índice; 4 elaboración de una planilla con los datos de cada provincia correspondientes a las variables elegidas y al valor porcentual provincial de cada índice, obtenido mediante la aplicación de la respectiva fórmula; 5 identificación de tres categorías continuas para cada índice, y 6 definición de la taxonomía. Resultados. De este modo, se obtuvo un mapa nacional de la situación de salud materna y de la capacidad instalada en cada una de las 112 provincias bolivianas, lo cual permitió seleccionar un número mínimo de provincias para realizar intervenciones conjuntas entre el Ministerio de Salud y Previsión Social y otras instituciones. Las 9 provincias seleccionadas tienen un total de 26 municipios que implican a 17 distritos sanitarios y concentran el 29% de la población del país, el 33% de las muertes maternas y el 35% de las defunciones neonatales tempranas estimadas. Conclusiones. Basándose en información disponible, el método utilizado proporcionó un mapa de la situación global, en este caso de la salud materna, de las 112 provincias de los 9 departamentos bolivianos que permite identificar territorios críticos para las intervenciones en materia de salud.

  5. Air pollution biomonitoring using terrestrial mosses; Il biomonitoraggio di pollutanti nell`aria mediante i muschi

    Energy Technology Data Exchange (ETDEWEB)

    Cenci, R. M. [C.C.R. Ispra, Istituto dell`Ambiente, La Spezia (Italy); Palmieri, F.; Mousty, F. [U.S.L. n.5, La Spezia (Italy)

    1998-06-01

    The spatial-temporal concentration of elements harmful for life has been investigated using terrestrial mosses of the Hypnum cupressiforme species. The results obtained have been implemented with elements of soil collected in the same area, for a wider information. Concerning the mosses, the investigated elements are: Al, As, Cd, Co, Cr, Fe, Mn, Pb, Cu, Ti, V, and Zn. The elements investigated on soil samples, collected only once, are: As, Cd, Cr, Mn, Ni, Cu and Pb. The results obtained allowed to identify an area of about 100 Kmq, located North-West with regard to La Spezia city (Italy), where the concentrations are higher than the remaining area. In the smaller investigation area, a surface of 0,12 kmq was found where the fall out rate for Pb is 0,21 g m-2 y-1, a value three times greater than the maximal european value tested in Rumania area (Ruehling, 1994). Mosses, which are highly efficient indicators of atmospheric pollution, have permitted to identify anthropogenic polluted areas and to evaluate the fall out rate. [Italiano] Mediante l`utilizzo del muschio appartenente alla specie Hypnum cupressiforme e` stata valutata la distribuzione nello spazio e nel tempo della concentrazione di elementi dannosi per la salute dell`uomo. I dati raccolti sono stati ampliati analizzando campioni di suolo raccolti nelle medesime localita`. Complessivamente sono state approntate 32 stazioni. Dopo il posizionamento i talli di muschio venivano raccolti per tre volte con cadenza quadrimestrale. per quanto riguarda i muschi sono state integrate le concentrazioni dei seguenti elementi: Al, As, Cd, Co, Cr, Fe, Mn, Pb, Cu, Ti, V, e Zn. Nei suoli, raccolti una sola volta, l`elenco degli elementi e` stato il seguente: As, Cd, Cr, Mn, Ni, Cu e Pb. I risultati ottenuti hanno permesso l`identificazione di un`area di circa 100 kmq posta a Nord-Ovest della citta` di La Spezia, dove le concentrazioni di As, Co, Ni, Cu, Ti e V sono superiori rispetto alla restante area. Nella piccola area

  6. Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of Japanese encephalitis virus

    International Nuclear Information System (INIS)

    Huang, S-H; Tsai, M-H; Lin, C-W; Yang, T-C; Chuang, P-H; Tsai, I-S; Lu, H-C; Wan Lei; Lin, Y-J; Lai, C-H

    2008-01-01

    Virus isolation and antibody detection are routinely used for diagnosis of Japanese encephalitis virus (JEV) infection, but the low level of transient viremia in some JE patients makes JEV isolation from clinical and surveillance samples very difficult. We describe the use of gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of JEV from its RNA genome. We tested the effect of gold nanoparticles on four different PCR systems, including conventional PCR, reverse-transcription PCR (RT-PCR), and SYBR green real-time PCR and RT-PCR assays for diagnosis in the acute phase of JEV infection. Gold nanoparticles increased the amplification yield of the PCR product and shortened the PCR time compared to the conventional reaction. In addition, nanogold-based real-time RT-PCR showed a linear relationship between Ct and template amount using ten-fold dilutions of JEV. The nanogold-based RT-PCR and real-time quantitative RT-PCR assays were able to detect low levels (1-10 000 copies) of the JEV RNA genomes extracted from culture medium or whole blood, providing early diagnostic tools for the detection of low-level viremia in the acute-phase infection. The assays described here were simple, sensitive, and rapid approaches for detection and quantitation of JEV in tissue cultured samples as well as clinical samples

  7. DNA fingerprinting by ERIC-PCR for comparing Listeria spp. strains isolated from different sources in San Luis: Argentina Caracterización molecular por ERIC-PCR de cepas de Listeria spp. aisladas de diversos orígenes en San Luis: Argentina

    Directory of Open Access Journals (Sweden)

    A. Laciar

    2006-04-01

    Full Text Available In this study, a total of 24 Listeria spp. strains were analyzed. Twenty-two isolates were obtained in San Luis (Argentina from human, animal, and food samples. Two types of strains, Listeria monocytogenes CLIP 22762 and Listeria innocua CLIP 74915, were included as reference strains. All isolates were biochemically identified and characterized by serotyping, phage typing, and amplification of the flaA gene by polymerase chain reaction (PCR. Repetitive intergenic consensus (ERIC sequence-based PCR was used to generate DNA fingerprints. On the basis of ERIC-PCR fingerprints, Listeria spp. strains were divided into three major clusters matching origin of isolation. ERIC-PCR fingerprints of human and animal isolates were different from those of food isolates. In addition, groups I and II included ten L. monocytogenes strains, and only one Listeria seeligeri strain. Group III included nine L. innocua strains and four L. monocytogenes strains. Computer evaluation of ERIC-PCR fingerprints allowed discrimination between the tested serotypes 1/2b, 4b, 6a, and 6b within each major cluster. The index of discrimination calculated was 0.94. This study suggests that the ERIC-PCR technique provides an alternative method for the identification of Listeria species and the discrimination of strains within one species.En este estudio se analizaron 24 cepas de Listeria spp. De ellas, 22 fueron obtenidas en San Luis (Argentina, a partir de muestras humanas, de animales y alimentos. Se incluyeron 2 cepas de referencia Listeria monocytogenes CLIP 22762 y Listeria innocua CLIP 74915. Todos los aislamientos fueron identificados bioquímicamente y caracterizados por serotipificación, fagotipificación y detección del gen flaA por reacción en cadena de la polimerasa (PCR. Se generaron perfiles de bandas de ADN mediante la amplificación de secuencias repetitivas de consenso intergénico de enterobacterias (ERIC-PCR. De acuerdo a los resultados obtenidos por ERIC-PCR

  8. Designing multiplex PCR system of Campylobacter jejuni for efficient typing by improving monoplex PCR binary typing method.

    Science.gov (United States)

    Yamada, Kazuhiro; Ibata, Ami; Suzuki, Masahiro; Matsumoto, Masakado; Yamashita, Teruo; Minagawa, Hiroko; Kurane, Ryuichiro

    2015-01-01

    Campylobacter jejuni is responsible for the majority of Campylobacter infections. As the molecular epidemiological study of outbreaks, pulsed-field gel electrophoresis (PFGE) is performed in general. But PFGE has several problems. PCR binary typing (P-BIT) method is a typing method for Campylobacter spp. that was recently developed, and was reported to have a similar discriminatory power and stability to those of PFGE. We modified the P-BIT method from 18 monoplex PCRs to two multiplex PCR systems (mP-BIT). The same results were obtained from monoplex PCRs using original primers and multiplex PCR in the representative isolates. The mP-BIT can analyze 48 strains at a time by using 96-well PCR systems and can identify C. jejuni because mP-BIT includes C. jejuni marker. The typing of the isolates by the mP-BIT and PFGE demonstrated generally concordant results and the mP-BIT method (D = 0.980) has a similar discriminatory power to that of PFGE with SmaI digest (D = 0.975) or KpnI digest (D = 0.987) as with original article. The mP-BIT method is quick, simple and easy, and comes to be able to perform it at low cost by having become a multiplex PCR system. Therefore, the mP-BIT method with two multiplex PCR systems has high potential for a rapid first-line surveillance typing assay of C. jejuni and can be used for routine surveillance and outbreak investigations of C. jejuni in the future. Copyright © 2014 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  9. Síntesis de nitruro de titanio mediante láser y energía solar concentrada

    Directory of Open Access Journals (Sweden)

    García, I.

    1998-04-01

    Full Text Available The possibility of the employment of solar energy concentrated by Fresnel lens is investigated in order to synthesize materials by gas-solid reaction. These first results are compared by two similar techniques as high power laser and xenon are lamp. The TiN coatings obtained with xenon are lamp and Fresnel lens are homogenous, without pores or defects, with a uniform thickness of about 6 μm for treatments of 2 min. The good quality of the TiN coating for all the testing conditions was confirmed by the x-ray diffraction measurements.

    Se presenta la utilización de la energía solar concentrada mediante lentes de Fresnel para la síntesis de materiales por reacción gas-sólido. Estos primeros resultados sobre nitruración superficial de titanio y aleaciones de titanio se comparan con los obtenidos con técnicas similares como el láser de alta potencia y la lámpara de descarga de xenón. Las capas de nitruro de titanio obtenidas mediante energía solar concentrada por lentes de Fresnel y lámpara de xenón son homogéneas, sin grietas ni defectos, y con un espesor uniforme de 6 μm en tiempos de sólo 2 min. La buena calidad de estas capas se confirma mediante difracción de rayos X.

  10. Detection and subtyping (H5 and H7) of avian type A influenza virus by reverse transcription-PCR and PCR-ELISA

    DEFF Research Database (Denmark)

    Munch, M.; Nielsen, L.P.; Handberg, Kurt

    2001-01-01

    A. A panel of reference influenza strains from various hosts including avian species, human, swine and horse were evaluated in a one tube RT-PCR using primers designed for the amplification of a 218 bp fragment of the NP gene. The PCR products were detected by PCR-ELISA by use of an internal......Avian influenza virus infections are a major cause of morbidity and rapid identification of the virus has important clinical, economical and epidemiological implications. We have developed a one-tube Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) for the rapid diagnosis of avian influenza...... catching probe confirming the NP influenza A origin. The PCR-ELISA was about 100 times more sensitive than detection of PCR products by agarose gel electrophoresis. RT-PCR and detection by PCR-ELISA is comparable in sensitivity to virus propagation in eggs. We also designed primers for the detection...

  11. One-step triplex PCR/RT-PCR to detect canine distemper virus, canine parvovirus, and canine kobuvirus.

    Science.gov (United States)

    Liu, Dafei; Liu, Fei; Guo, Dongchun; Hu, Xiaoliang; Li, Zhijie; Li, Zhigang; Ma, Jianzhang; Liu, Chunguo

    2018-01-23

    To rapidly distinguish Canine distemper virus (CDV), canine parvovirus (CPV), and canine kobuvirus (CaKoV) in practice, a one-step multiplex PCR/RT-PCR assay was developed, with detection limits of 10 2.1 TCID 50 for CDV, 10 1.9 TCID 50 for CPV and 10 3 copies for CaKoV. This method did not amplify nonspecific DNA or RNA from other canine viruses. Therefore, the assay provides a sensitive tool for the rapid clinical detection and epidemiological surveillance of CDV, CPV and CaKoV in dogs.

  12. A novel polymerase chain reaction (PCR) for rapid isolation of a new ...

    African Journals Online (AJOL)

    mediated self-formed panhandle PCR, for gene or chromosome walking. It combined the advantages of ligation-mediated PCR in its specificity and of panhandle PCR in its efficiency. Self-formed panhandle PCR was used for a new rbcS gene ...

  13. Evaluation of a PCR for detection of Actinobacillus pleuropneumoniae in mixed bacterial cultures from tonsils

    DEFF Research Database (Denmark)

    Gram, T.; Ahrens, Peter; Nielsen, J.P.

    1996-01-01

    strains of A. lignieresii. The lower detection limit of the PCR test was 10(3) A. pleuropneumoniae CFU/PCR test tube and was not affected by addition of 10(6) E. coli CFU/PCR test tube. Mixed bacterial cultures from tonsils of 101 pigs from 9 different herds were tested by culture and by PCR using four...

  14. Análisis del yeso empleado en revestimientos exteriores mediante técnicas geológicas

    OpenAIRE

    Sanz Arauz, David

    2011-01-01

    El presente trabajo se centra en el yeso fabricado en hornos tradicionales y empleado históricamente para la ejecución de revestimientos exteriores. Para ello se realiza un estudio documental y un trabajo experimental mediante técnicas geológicas. En el estudio documental se ha pasado revisión a la historia del yeso como material de construcción con especial atención a su empleo en revestimientos exteriores, desde la antigüedad hasta mitad del siglo XX, momento en el que la industrializaci...

  15. Estudio de la microestructura femoral de pacientes con coxartrosis y con fractura de cadera mediante micro-TAC

    OpenAIRE

    Sainz-Aja Guerra, J.A.; Alonso, M.A.; Ferreño Blanco, D.; Pérez-Núñez, M.I.; Ruiz Martínez, E.; García-Ibarbia, C.; Casado del Prado, J.A.; Gutiérrez-Solana, F.; Riancho, J.A.

    2016-01-01

    La disminución de la densidad mineral ósea (DMO), es decir, del volumen de tejido óseo por unidad de volumen del esqueleto, es característica de la osteoporosis, mientras que se ha sugerido que la artrosis se acompaña de un aumento de la DMO a nivel local y sistémico. Para comprobar esta hipótesis analizamos mediante microTAC el hueso trabecular de la cabeza femoral de 10 pacientes con fractura de cadera y 9 con coxartrosis. El análisis no reveló diferencias significativas entre ambos grupos ...

  16. Estudio de una celda de fabricación flexible mediante la simulación de eventos discretos

    OpenAIRE

    Sanz Lucero, Flavia

    2014-01-01

    En la actualidad las fábricas se ven afectadas por la alta competencia tanto nacional como internacional, y obligadas a mejorar sus procesos productivos para poder continuar siendo competitivas en un mercado cada vez más hostil. El presente proyecto trata de resolver una parte importante de esta mejora: la reducción de los tiempos improductivos o muertos, mediante la búsqueda de una secuencia óptima de alimentación en una celda de fabricación flexible compuesta por máquinas ali...

  17. Reducción de percloroetileno y cromo hexavalente mediante FE(0) y bioestimulación de microorganismos anaerobios

    OpenAIRE

    VÁZQUEZ MORILLAS, Alethia; VACA MIER, Mabel; BELTRÁN VILLAVICENCIO, Margarita; LÓPEZ CALLEJAS, Raymundo; ÁLVAREZ, Pedro J

    2007-01-01

    En este trabajo se estudiaron las interacciones Fe-microorganismos simulando el entorno de una barrera reactiva permeable (BRP), para la reducción de percloroetileno (PCE) y cromo hexavalente en acuíferos contaminados. La bioestimulación de los microorganismos se llevó a cabo mediante la adición de un cosustrato para el desarrollo de las comunidades microbianas presentes de manera natural en el suelo. Se empleó un compuesto comercial que libera hidrógeno (HRC, por sus siglas en inglés) a tasa...

  18. Estudio de aleaciones amorfas Hf1-x Cux mediante correlaciones angulares perturbadas

    OpenAIRE

    Damonte, Laura Cristina

    1988-01-01

    Con la intención de contribuir a un mejor entendimiento de la estructura atómica local en aleaciones del tipo MT-MT, en este trabajo de tesis se realiza un estudio, mediante la aplicación de la técnica TDPAC, sobre aleaciones amorfas Hf1-x Cux (x=0.33, 0.44, 0.50, 0.59) y sus contrapartes cristalinas. Muy pocos trabajos han sido publicados sobre este sistema. El objetivo general de este trabajo es analizar el orden local en estas aleaciones, su evolución con tratamientos térmicos (r...

  19. Protocolo para la producción de plantas dihaploides de trigo duro mediante cruzamientos con maíz

    OpenAIRE

    García Llamas, Carmen

    2011-01-01

    El objetivo de este trabajo ha sido desarrollar un protocolo para producir plantas dihaploides de trigo duro, mediante cruzamientos con maíz, que permita acortar la duración de los programas de mejora de esta especie. Entre las variantes de este método se ha elegido el cultivo de tallos cortados, porque permite controlar las condiciones ambientales durante el desarrollo de los embriones y abaratar los costes de aplicación de las hormonas. Se han abordado y mejorado tres aspectos de esta técni...

  20. Refuerzo de puentes por cambio de esquema estructural: optimización mediante algoritmo genético

    OpenAIRE

    Valenzuela, Matías A.; Casas Rius, Joan Ramon

    2011-01-01

    La presente comunicación entrega un estudio detallado del proceso constructivo y de tesado para el refuerzo de puentes con tipología longitudinal de vigas continuas convirtiéndolos en puentes en arco atirantado (tipo network). Se presentan las etapas básicas propuestas en aspectos de construcción, se establecen las hipótesis del estudio de tesado e implementa el método de tesado a partir de la optimización automatizada mediante algoritmos genéticos, definiendo criterios como: variables de ...

  1. Tratamiento de las osteitis diafisarias de tibia mediante sinostosis tibioperonea y resección ósea

    OpenAIRE

    Fernández Sabaté, Alfons; Riu Labrador, R.; Moreta Munujos, D.; Cáceres Palou, Enrique

    1984-01-01

    El tratamiento de las osteítis crónicas de la diáfisis tibial obliga a menudo a practicar una amplia resección para alcanzar el tejido óseo sano y esta exigencia operatoria debilita la diáfisis. Para obviar la fractura operatoria o posterior por fatiga y para facilitar la resección sin temor se ha utilizado una táctica quirúrgica consistente en una solidarización tibioperonea que construye un residente eje en la pierna ya sea mediante injerto intertibioperoneo ya con peroné protibia de Zanoli...

  2. Sistema “Gestor Fiducia Fondos JEE” Mediante Servicios de Cloud Computing, Estudio de Factibilidad e Implementación

    OpenAIRE

    Unda, Priscila; Coral, Henry; Marcillo, Diego

    2016-01-01

    Este proyecto presenta el estudio de factibilidad y el diseño e implementación del sistema “Gestor Fiducia Fondos JEE”, mediante servicios en la nube computacional. Para llevarlo a cabo, se realizó una evaluación técnica de tres importantes proveedores de nubes computacionales Amazon Ec2, Rackspace, y Terremark, valorando la calidad del soporte, costos, escalabilidad y usabilidad. Posteriormente, se realizó el diseño y definición de una infraestructura básica, media y superior que permita alb...

  3. Segmentación automática de noticias mediante procesamiento de formas prosódicas

    OpenAIRE

    Mas Manchón, Lluís

    2014-01-01

    La segmentación automatizada de noticias en tiempo real es un problema que ha tenido un abordaje fundamentalmente lingüístico y de procesamiento de la señal en los últimos años. El trabajo que presentamos tiene un enfoque sustancialmente diferente: desde una perspectiva comunicológica, se toman las formas prosódicas típicas del discurso informativo y se intentan programar mediante el procesamiento cepstrum en el entorno Labview. Después de numerosas pruebas se fijan los diferentes parámetros ...

  4. Generación de escenarios de planificación de redes mediante técnicas OLAP

    OpenAIRE

    Ramos García, Jesús

    2015-01-01

    RESUMEN: Debido a la gran cantidad de datos que a veces se tienen que manejar, es necesario el uso de herramientas que se encarguen de almacenar, reducir, extraer, analizar, procesar o modificar dichos datos de la forma más rápida y sencilla posible. Las bases de datos multidimensionales, con el nombre de cubo OLAP (On-Line Analytical Processing) se encargan de representar la información elegida de forma multidimensional, es decir, mediante cubos de varias dimensiones, haciendo que dicha info...

  5. Propuesta de mejora de habilidades sociales en niños con discapacidad intelectual mediante la terapia asistida por animales

    OpenAIRE

    Bermejo Lorenzo, Julia

    2015-01-01

    Este Trabajo de Fin de Grado se centra en la aplicación práctica de un programa de mejora de la Educación Socioemocional, utilizando como herramienta innovadora la Terapia Asistida con Animales de Compañía, aplicándose a un caso real de un alumno con Parálisis Cerebral y Discapacidad Intelectual asociada. Los objetivos de las actividades están centrados en el desarrollo de la autoconciencia, el autocontrol, la conciencia social y la toma de decisiones responsables. Mediante este proyecto se p...

  6. Metodologías de análisis de túneles excavados con tuneladora mediante el programa Plaxis

    OpenAIRE

    Alcahuaman Villanueva, Victor Abner

    2016-01-01

    El programa de elementos finitos Plaxis se ha convertido en una herramienta popular para diseñar y estudiar túneles excavados mediante máquinas tuneladoras. En el caso de túneles urbanos, los movimientos originados en la excavación son de gran importancia, y se ha observado que en función de la metodología de cálculo seguida en Plaxis los resultados pueden ser muy diferentes. La tesina pretende comparar las diversas metodologías disponibles y establecer criterios para su uso en casos reales....

  7. Análisis comparativo de distintas toolkits para el reconocimiento biométrico de personas mediante voz

    OpenAIRE

    Ruíz, Silvia; Miranda, Ernesto; Herlein, Mauro; Etchart, Graciela; Alvez, Carlos E.

    2017-01-01

    El objetivo de este trabajo es realizar un análisis comparativo de distintas toolkits para el reconocimiento biométrico de personas mediante voz. Hoy en día los sistemas de identificación de personas se han convertido en una necesidad para la sociedad. A medida que avanza la tecnología y la aplicación de la misma en entornos tanto de ocio como de seguridad, la evolución en desarrollo biométrico es muy grande. Los sistemas de identificación o verificación tradicionales (tarjetas o claves) se h...

  8. Generación de experiencias visuales en ciegos mediante estimulación táctil repetitiva

    OpenAIRE

    Tomás Ortiz; Juan M. Santos

    2012-01-01

    Estudios recientes de nuestro laboratorio han establecido que se puede generar una activación estable del córtex visual en ciegos mediante entrenamiento táctil pasivo prolongado. Esta neuroplasticidad cortical se acompaña en un 40% de los participantes invidentes de sensaciones visuales, así como de una mayor rapidez en el reconocimiento táctil de información espacial tales como líneas, iconos o imágenes, y es crucial para su correcta interpretación. Estos hallazgos pueden ser útiles para el ...

  9. Detección de movimiento mediante técnicas radar CW-FM en banda W

    OpenAIRE

    Vargas González, Daniel

    2014-01-01

    Aplicació de diverses tècniques de detecció i localització per detectar moviments amb un radar C2-FM que opera en banda W. [ANGLÈS] Integration of a SAR adquisition system using a FM-CW 94 GHz radar and test the system by different measurement campaigns with the aim of detecting micrometic displacements using a phase analysis of the recived signal [CASTELLÀ[ Integración de un sistema de adquisición SAR mediante el uso de un radar FM-CW a 94 GHz y probar la validez del mencionado sistema...

  10. Recubrimientos metálicos sobre alúmina mediante procesos de reducción autocatalítica

    Directory of Open Access Journals (Sweden)

    Gómez de Salazar, J. M.

    2000-10-01

    Full Text Available In this work, a method for obtaining copper coating on alumina is described. One of the main applications for this coating is in the electronic industry, although it can be used as well as interlayer for dissimilar bonding between metals and alumina, both by solid state joining and by active brazing. The optimal activation conditions for the alumina using Ni salts and the influence of the surface preparation on the copper coating characteristic are described. The coating application is based on an autocatalithic reduction method. The influence of the deposition rate on the adherence of the copper coating has been studied as well. A kinetic study was carried out applying gravimetric and electrochemical methods. To obtaining coating with high adherence, it was necessary to apply heat treatments after the metallization process. The main objective of them was to achieve a chemical bond between the alumina substrate and the copper coating by formation of Al-Cu spinels, instead of the single mechanical bond.

    En el presente trabajo se describe un método de obtención de recubrimientos de cobre sobre un cerámico tenaz como es la alúmina. Una de sus principales aplicaciones se encuentra en la industria electrónica, aunque también puede ser empleado como intermediario en la fabricación de uniones disimilares entre un metal y un cerámico mediante técnicas de unión en estado sólido o en soldadura fuerte reactiva. Se describen las condiciones óptimas de activación de la alúmina mediante sales de Ni, y la influencia que posee la preparación superficial de este substrato (Al2O3 sobre las capas de Cu obtenidas. Este proceso se realiza mediante reducción autocatalítica, habiéndose estudiado como influye la velocidad de deposición sobre la adherencia de la capa de cobre. También se realizaron estudios cinéticos del proceso de recubrimiento mediante ensayos gravimétricos y electroquímicos. Con el fin de obtener recubrimientos que

  11. Control estadístico de la calidad de un servicio mediante Gráficas X y R

    OpenAIRE

    Alberto Isaac Pierdant Rodríguez; Jesús Rodríguez Franco

    2009-01-01

    Las empresas y organismos públicos que proporcionan servicios en México no utilizan frecuentemente técnicas cuantitativas para el control de calidad de dicho servicio, por lo que este trabajo representa una propuesta de control de calidad mediante herramientas simples de control estadístico de calidad. Existen diversas técnicas cualitativas y pocas técnicas cuantitativas como las gráficas, que nos permiten determinar si la prestación de un servicio se encuentra bajo control; es decir, verific...

  12. Control de bacteriemia nosocomial pediátrica mediante un programa de cultivo de soluciones parenterales en uso

    OpenAIRE

    Muñoz Juan M.; Macías Alejandro E.; Guerrero Francisco J.; Hernández Isabel; Medina Humberto; Vargas Enrique

    1999-01-01

    OBJETIVO. Dado que Klebsiella, Enterobacter y Serratia se multiplican en soluciones parenterales y son responsables de una elevada proporción de bacteriemias en los hospitales de México, se propone una estrategia de control mediante la vigilancia microbiológica de las soluciones en uso. MATERIAL Y MÉTODOS. Hospital de enseñanza de segundo nivel con 193 camas. Atiende principalmente pacientes de escasos recursos. En 1992 se inició la vigilancia de la esterilidad de las soluciones parenterales ...

  13. Teleoperación de un robot mindstorm mediante técnicas de visión artificial

    OpenAIRE

    Méndez Rodríguez, Eduardo

    2011-01-01

    Las tres partes tratadas son la teleoperacióon (principalmente con comunicaciones mediante bluetooth), la visión arti cial para elementos móviles en tiempo real y proyectos realizados con el Mindstorm NXT. Hay que destacar que en los dos primeros campos hay innumerables aplicaciones debido a que ambos campos siguen hoy en día bajo invetigación en busca de lograr mejoras tecnológicas, también los proyectos con el Mindstorm están creciendo exponencialmente, mayormente debido a...

  14. Programa Antiestrés de Sincronización Cerebral mediante estimulación visual

    OpenAIRE

    Riccardi Sabatier, Yanitza; Caraballo Pons, Isabel; Miyares Cao, Carlos Manuel; Lago Mendoza, Guillermo; Lauzán Álvarez, Efreín

    2014-01-01

    Se presenta un programa computarizado que permite la sincronización de las ondas cerebrales mediante una estimulación visual, a una frecuencia similar a la actividad eléctrica del cerebro en estado de sedación, lo cual favorece la disminución de los niveles de estrés en los pacientes que acuden al Centro de Histoterapia Placentaria para tratar las patologías de Vitiligo, Psoriasis y Alopecia Areata. La aplicación informática fue programada en lenguaje Delphi 7.0 y cuenta con dos módulos para ...

  15. Educación mediante enciclopedias visuales temáticas: INTYPEDIA un caso de éxito.

    OpenAIRE

    Ramió Aguirre, Jorge; Muñoz Muñoz, Alfonso

    2013-01-01

    La presente investigación recoge una experiencia de innovación educativa que demuestra la utilidad de las enciclopedias visuales temáticas para la formación académica a través de Internet. Los objetivos de esta experiencia recaen en verificar si es posible crear enciclopedias visuales temáticas que mediante recursos audiovisuales permitan el aprendizaje de aspectos complejos de una manera asequible. Con el interés de obtener datos que permitan comprobar la idoneidad de este tipo de recursos f...

  16. Análisis del yeso empleado en revestimientos exteriores mediante técnicas geológicas

    OpenAIRE

    Sanz Arauz, David

    2009-01-01

    El presente trabajo se centra en el yeso fabricado en hornos tradicionales y empleado históricamente para la ejecución de revestimientos exteriores. Para ello se realiza un estudio documental y un trabajo experimental mediante técnicas geológicas. En el estudio documental se ha pasado revisión a la historia del yeso como material de construcción con especial atención a su empleo en revestimientos exteriores, desde la antigüedad hasta mitad del siglo XX, momento en el que la industrializaci...

  17. Pronóstico del IPC de la Bolsa Mexicana de Valores Mediante el Uso de Reglas y Redes Neuronales

    OpenAIRE

    Cázares Carrillo, Juan G.

    2004-01-01

    La presente investigación es un esfuerzo para el desarrollo de un pronosticador con resultados aceptables para el mercado de valores destinándose su uso como una herramienta de apoyo a la toma de decisiones financieras. El pronóstico se realiza mediante la identificación de patrones utilizando como herramientas técnicas de Inteligencia Artificial, específicamente Redes Neuronales de Retropropagación y Algoritmos Genéticos. Las principales aportaciones de la presente investigación son: la dete...

  18. Eliminación de contaminantes emergentes en aguas residuales mediante oxidación avanzada con ozono y ultrasonidos

    OpenAIRE

    Abellán Soler, Manuel; Lardín Mifsut, Carlos; Morales Cavero, Eduardo; Pastor Alcañiz, Laura; Martínez Muro, Juan Luis; Santos Asensi, José María; Ibáñez Martínez, María; Hernández Hernández, Félix

    2013-01-01

    Algunos contaminantes emergentes, principalmente fármacos de diferentes clases así como drogas de abuso, pueden estar presentes en las aguas residuales urbanas, no siendo posible su eliminación mediante las técnicas convencionales de depuración. Se ha realizado un estudio en planta piloto en dos estaciones depuradoras de aguas residuales (EDAR), Font de la Pedra (Muro de Alcoy) y Molina de Segura (Murcia), con el fin de determinar la eficacia de eliminación de ciertos ...

  19. Typing of Mycoplasma pneumoniae by PCR-mediated DNA fingerprinting

    NARCIS (Netherlands)

    Ursi, D; Ieven, M; van Bever, H; Quint, W; Niesters, H G; Goossens, H

    PCR fingerprinting was used to characterize clinical isolates of Mycoplasma pneumoniae. Among 24 strains tested, two types were distinguished. Nineteen strains belonged to type 1, whereas only 5 strains belonged to type 2. The majority of strains isolated since 1991 in Belgium belong to type 1. No

  20. Halal authenticity of gelatin using species-specific PCR.

    Science.gov (United States)

    Shabani, Hessam; Mehdizadeh, Mehrangiz; Mousavi, Seyed Mohammad; Dezfouli, Ehsan Ansari; Solgi, Tara; Khodaverdi, Mahdi; Rabiei, Maryam; Rastegar, Hossein; Alebouyeh, Mahmoud

    2015-10-01

    Consumption of food products derived from porcine sources is strictly prohibited in Islam. Gelatin, mostly derived from bovine and porcine sources, has many applications in the food and pharmaceutical industries. To ensure that food products comply with halal regulations, development of valid and reliable analytical methods is very much required. In this study, a species-specific polymerase chain reaction (PCR) assay using conserved regions of mitochondrial DNA (cytochrome b gene) was performed to evaluate the halal authenticity of gelatin. After isolation of DNA from gelatin powders with known origin, conventional PCR using species-specific primers was carried out on the extracted DNA. The amplified expected PCR products of 212 and 271 bp were observed for porcine and bovine gelatin, respectively. The sensitivity of the method was tested on binary gelatin mixtures containing 0.1%, 1%, 10%, and 100% (w/w) of porcine gelatin within bovine gelatin and vice versa. Although most of the DNA is degraded due to the severe processing steps of gelatin production, the minimum level of 0.1% w/w of both porcine and bovine gelatin was detected. Moreover, eight food products labeled as containing bovine gelatin and eight capsule shells were subjected to PCR examination. The results showed that all samples contained bovine gelatin, and the absence of porcine gelatin was verified. This method of species authenticity is very useful to verify whether gelatin and gelatin-containing food products are derived from halal ingredients. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Comparison of polymerase chain reaction (PCR) and loop-mediated ...

    African Journals Online (AJOL)

    Comparison of polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) for diagnosis of Fusarium solani in human immunodeficiency virus (HIV) positive patients. ... The test was carried out in 1 h reaction at 65°C in a heater block. The specificity of the test was 100% and its sensitivity was a ...

  2. Introducing Undergraduate Students to Real-Time PCR

    Science.gov (United States)

    Hancock, Dale; Funnell, Alister; Jack, Briony; Johnston, Jill

    2010-01-01

    An experiment is conducted, which in four 3 h laboratory sessions, introduces third year undergraduate Biochemistry students to the technique of real-time PCR in a biological context. The model used is a murine erythroleukemia cell line (MEL cells). These continuously cycling, immature red blood cells, arrested at an early stage in erythropoiesis,…

  3. A Trio of Human Molecular Genetics PCR Assays

    Science.gov (United States)

    Reinking, Jeffrey L.; Waldo, Jennifer T.; Dinsmore, Jannett

    2013-01-01

    This laboratory exercise demonstrates three different analytical forms of the polymerase chain reaction (PCR) that allow students to genotype themselves at four different loci. Here, we present protocols to allow students to a) genotype a non-coding polymorphic Variable Number of Tandem Repeat (VNTR) locus on human chromosome 5 using conventional…

  4. Fuel utilization in a progressive conversion reactor (PCR)

    International Nuclear Information System (INIS)

    Leyse, C.F.; Judd, J.L.

    1981-05-01

    Preliminary studies indicate that for once-through fuel cycles, the PCR offers potential improvements over current LWRs in the following major areas: improved uranium utilization (reduced uranium demand), degraded plutonium product in spent fuel, reduced plutonium content of spent fuel, reduced amount of spent fuel, reduced fissile content of spent fuel, and reduced separative work

  5. Statistical aspects of quantitative real-time PCR experiment design

    Czech Academy of Sciences Publication Activity Database

    Kitchen, R.R.; Kubista, Mikael; Tichopád, Aleš

    2010-01-01

    Roč. 50, č. 4 (2010), s. 231-236 ISSN 1046-2023 R&D Projects: GA AV ČR IAA500520809 Institutional research plan: CEZ:AV0Z50520701 Keywords : Real-time PCR * Experiment design * Nested analysis of variance Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.527, year: 2010

  6. Design and Optimization of Reverse-Transcription Quantitative PCR Experiments

    Czech Academy of Sciences Publication Activity Database

    Tichopád, A.; Kitchen, R.; Riedmaier, I.; Becker, Ch.; Ståhlberg, A.; Kubista, Mikael

    2009-01-01

    Roč. 55, č. 10 (2009), s. 1816-1823 ISSN 0009-9147 Institutional research plan: CEZ:AV0Z50520701 Keywords : Design * optimization * RT qPCR Subject RIV: EG - Zoology Impact factor: 6.263, year: 2009

  7. Comparison of conventional culture and real-time quantitative PCR ...

    African Journals Online (AJOL)

    2009-10-28

    Oct 28, 2009 ... for each sample), which can be used to determine the success of the PCR reaction ... good performance in the absence of an internal control. First, a. GenBank query ..... the context of a health risk management programme.

  8. Bioinformatic tools and guideline for PCR primer design | Abd ...

    African Journals Online (AJOL)

    Bioinformatics has become an essential tool not only for basic research but also for applied research in biotechnology and biomedical sciences. Optimal primer sequence and appropriate primer concentration are essential for maximal specificity and efficiency of PCR. A poorly designed primer can result in little or no ...

  9. Typing of Mycoplasma pneumoniae by PCR-mediated DNA fingerprinting

    NARCIS (Netherlands)

    Ursi, D; Ieven, M; van Bever, H; Quint, W; Niesters, H G; Goossens, H

    1994-01-01

    PCR fingerprinting was used to characterize clinical isolates of Mycoplasma pneumoniae. Among 24 strains tested, two types were distinguished. Nineteen strains belonged to type 1, whereas only 5 strains belonged to type 2. The majority of strains isolated since 1991 in Belgium belong to type 1. No

  10. Real-time PCR for Strongyloides stercoralis-associated meningitis.

    Science.gov (United States)

    Nadir, Eyal; Grossman, Tamar; Ciobotaro, Pnina; Attali, Malka; Barkan, Daniel; Bardenstein, Rita; Zimhony, Oren

    2016-03-01

    Four immunocompromised patients, immigrants from Ethiopia, presented with diverse clinical manifestations of meningitis associated with Strongyloides stercoralis dissemination as determined by identification of intestinal larvae. The cerebrospinal fluid of 3 patients was tested by a validated (for stool) real-time PCR for S. stercoralis and was found positive, establishing this association. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Role of Polymerase Chain Reaction (PCR) in the detection of ...

    African Journals Online (AJOL)

    Rajeh Ali

    2014-06-20

    Jun 20, 2014 ... in 2013. Objectives: This study aimed to investigate S. aureus in some clinical samples by PCR and study .... culture, the isolates of S. aureus were incubated at 37 °C for. 18–24 h .... characteristics, and ability to form biofilm.

  12. Role of Polymerase Chain Reaction (PCR) in the detection of ...

    African Journals Online (AJOL)

    Background: Staphylococcus aureus is mainly acquired from hospital infections and demonstrated the ability of developing resistance to many antibiotics. Polymerase Chain Reaction (PCR) was used to identify antibiotic-resistant isolates. This study was conducted in Al-Mujtahed, Al-Mouwasat and the Children Hospitals in ...

  13. Polymerase Chain Reaction (PCR) provides a superior tool for the ...

    African Journals Online (AJOL)

    Polymerase Chain Reaction (PCR) provides a superior tool for the diagnosis of Pneumococcal Infection in Burkina Faso. Y Chaibou, M Congo/Ouedraogo, I Sanou, H Somlare, K Ouattara, CM Kienou, H Belem, E Sampo, SA Traore, R Traore/Ouedraogo, C Hatcher, L Mayer, X Wang, L Sangare ...

  14. Comparison of agglutination test, microscopy and nPCR for ...

    African Journals Online (AJOL)

    action (nPCR) for the detection of T. gondii infection in mice (n=399) inoculated with heart .... Direct Agglutination Test (DAT) (Toxo screen DA, Biomerieux®, France) fol- ... 0.3 µl external forward Primer (50 µM), 0.3 µl external reverse Primer.

  15. Development of a multiplex PCR assay detecting 52 autosomal SNPs

    DEFF Research Database (Denmark)

    Sanchez Sanchez, Juan Jose; Phillips, C.; Børsting, Claus

    2006-01-01

    for amplifying 52 genomic DNA fragments, each containing one SNP, in a single tube, and accurately genotyping the PCR product mixture using two single base extension reactions. This multiplex approach reduces the cost of SNP genotyping and requires as little as 0.5 ng of genomic DNA to detect 52 SNPs. We used...

  16. How useful is PCR in the diagnosis of malaria?

    NARCIS (Netherlands)

    Hänscheid, Thomas; Grobusch, Martin P.

    2002-01-01

    Polymerase chain reaction (PCR) assays are the most sensitive and specific method to detect malaria parasites, and have acknowledged value in research settings. However, the time lag between sample collection, transportation and processing, and dissemination of results back to the physician limits

  17. PCR-DGGE fingerprints of microbial successional changes during ...

    African Journals Online (AJOL)

    PCR-DGGE fingerprints of microbial successional changes during fermentation of cereal-legume weaning foods. ... African Journal of Biotechnology ... Phenotypic identification and monitoring of the dynamics of naturally occurring microbial community responsible for the spontaneous fermentation of different cereal-legume ...

  18. FTA card utility for PCR detection of Mycobacterium leprae.

    Science.gov (United States)

    Aye, Khin Saw; Matsuoka, Masanori; Kai, Masanori; Kyaw, Kyaw; Win, Aye Aye; Shwe, Mu Mu; Thein, Min; Htoo, Maung Maung; Htoon, Myo Thet

    2011-01-01

    The suitability of the FTA® elute card for the collection of slit skin smear (SSS) samples for PCR detection of Mycobacterium leprae was evaluated. A total of 192 SSS leprosy samples, of bacillary index (BI) 1 to 5, were collected from patients attending two skin clinics in Myanmar and preserved using both FTA® elute cards and 70% ethanol tubes. To compare the efficacy of PCR detection of DNA from each BI class, PCR was performed to amplify an M. leprae-specific repetitive element. Of the 192 samples, 116 FTA® elute card and 112 70% ethanol samples were PCR positive for M. leprae DNA. When correlated with BI, area under the curve (AUC) values of the respective receiver-operating characteristic curves were similar for the FTA® elute card and ethanol collection methods (AUC=0.6). Taken together, our results indicate that the FTA® elute card, which enables the collection, transport, and archiving of clinical samples, is an attractive alternative to ethanol preservation for the detection of M. leprae DNA.

  19. Pure chromosome-specific PCR libraries from single sorted chromosomes

    NARCIS (Netherlands)

    VanDevanter, D. R.; Choongkittaworn, N. M.; Dyer, K. A.; Aten, J. A.; Otto, P.; Behler, C.; Bryant, E. M.; Rabinovitch, P. S.

    1994-01-01

    Chromosome-specific DNA libraries can be very useful in molecular and cytogenetic genome mapping studies. We have developed a rapid and simple method for the generation of chromosome-specific DNA sequences that relies on polymerase chain reaction (PCR) amplification of a single flow-sorted

  20. Development of a competitive PCR assay for the quantification of ...

    African Journals Online (AJOL)

    ONOS

    2010-01-25

    Jan 25, 2010 ... quantification of total Escherichia coli DNA in water. Omar Kousar Banu, Barnard .... Thereafter the product was ligated into the pGEM®T-easy cloning ... agarose gel using the high pure PCR product purification kit. (Roche® ...

  1. QUANTITATIVE PCR OF SELECTED ASPERGILLUS, PENICILLIUM AND PAECILOMYCES SPECIES

    Science.gov (United States)

    A total of 65 quantitative PCR (QPCR) assays, incorporating fluorigenic 5' nuclease (TaqMan®) chemistry and directed at the nuclear ribosomal RNA operon, internal transcribed spacer regions (ITS1 or ITS2) was developed and tested for the detection of Aspergillus, Penicillium and ...

  2. Diagnostic PCR: Comparative sensitivity of four probe chemistries

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Löfström, Charlotta; Sommer, Helle Mølgaard

    2009-01-01

    Three probe chemistries: locked nucleic acid (LNA), minor groove binder (MGB) and Scorpion were compared with a TaqMan probe in a validated real-time PCR assay for detection of food-borne thermotolerant Campylobacter. The LNA probe produced significantly lower Ct-values and a higher proportion of...

  3. Evaluation of nested PCR in diagnosis of fungal rhinosinusitis.

    Science.gov (United States)

    Badiee, Parisa; Gandomi, Behrooz; Sabz, Gholamabbass; Khodami, Bijan; Choopanizadeh, Maral; Jafarian, Hadis

    2015-02-01

    Given the importance of rapid diagnosis for fungal rhinosinusitis, this study aimed to evaluate the use of nested PCR to identify Aspergillus and Mucor species in clinical samples from patients with suspected fungal rhinosinusitis. Functional endoscopic sinus surgery specimens were collected from 98 patients with rhinosinusitis from 2012 to 2013. All samples were ground and cultured on sabouraud dextrose agar. The isolated fungi were identified based on their macroscopic and microscopic features. Fungal DNA was extracted from the tissue samples and nested PCR was performed with two sets of primers for Mucor and Aspergillus. Direct microscopic showed that 5.1% contained fungal components and 9.2% exhibited growth of fungi in culture. The most common agents isolated were Aspergillus fumigatus (n= 3), Aspergillus flavus (n=2), Penicillium sp (n=3) and Alternaria sp. (n=1). Mucor sp. was identified in the pathology smear from 1 patient. Positive results for fungal rhinosinusitis were obtained for a total of 10.2% by culture or pathology smear. Positive PCR results were obtained in 72 samples for Aspergillus and 31 samples for Mucor. Our results suggest that endoscopic sinus surgery specimens are not suitable for nested PCR, probably because of the accumulation of fungi that contaminate the environmental air. This drawback is a limiting factor for diagnosis with nasal cavity specimens. Therefore, molecular methods and conventional culture techniques are helpful complementary diagnostic methods to detect fungal rhinosinusitis and determine appropriate management for these patients.

  4. Quality control for quantitative PCR based on amplification compatibility test

    Czech Academy of Sciences Publication Activity Database

    Tichopád, Aleš; Bar, T.; Pecen, Ladislav; Kitchen, R.R.; Kubista, Mikael; Pfaffl, M.W.

    2010-01-01

    Roč. 50, č. 4 (2010), s. 308-312 ISSN 1046-2023 R&D Projects: GA AV ČR IAA500520809; GA AV ČR IAA500970904 Institutional research plan: CEZ:AV0Z50520701 Keywords : Quantitative PCR * Quality control * Amplification efficiency Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.527, year: 2010

  5. Comparison of the performance in detection of HPV infections between the high-risk HPV genotyping real time PCR and the PCR-reverse dot blot assays.

    Science.gov (United States)

    Zhang, Lahong; Dai, Yibei; Chen, Jiahuan; Hong, Liquan; Liu, Yuhua; Ke, Qiang; Chen, Yiwen; Cai, Chengsong; Liu, Xia; Chen, Zhaojun

    2018-01-01

    A new multiplex real-time PCR assay, the high-risk HPV genotyping real time PCR assay (HR HPV RT-PCR), has been developed to detect 15 high-risk HPV types with respective viral loads. In this report, a total of 684 cervical specimens from women diagnosed with vaginitis were assessed by the HR HPV RT-PCR and the PCR reaction and reverse dot blot (PCR-RDB) assays, using a PCR-sequencing method as a reference standard. A total coincidence of 97.7% between the HR HPV RT PCR and the PCR-RDB assays was determined with a Kappa value of 0.953. The HR HPV RT PCR assay had sensitivity, specificity, and concordance rates (accuracy) of 99.7%, 99.7%, and 99.7%, respectively, as confirmed by PCR-sequencing, while the PCR-RDB assay had respective rates of 98.8%, 97.1%, and 98.0%. The overall rate of HPV infection, determined by PCR-sequencing, in women diagnosed with vaginitis was 49.85%, including 36.26% of single infection and 13.6% of multiple infections. The most common infections among the 15 high-risk HPV types in women diagnosed with vaginitis were HPV-52, HPV-16, and HPV-58, with a total detection rate of 10.23%, 7.75%, and 5.85%, respectively. We conclude that the HR HPV RT PCR assay exhibits better clinical performance than the PCR-RDB assay, and is an ideal alternative method for HPV genotyping. In addition, the HR HPV RT PCR assay provides HPV DNA viral loads, and could serve as a quantitative marker in the diagnosis and treatment of single and multiple HPV infections. © 2017 Wiley Periodicals, Inc.

  6. Solid-phase PCR for rapid multiplex detection of Salmonella spp. at the subspecies level, with amplification efficiency comparable to conventional PCR

    DEFF Research Database (Denmark)

    Chin, Wai Hoe; Sun, Yi; Høgberg, Jonas

    2017-01-01

    Solid-phase PCR (SP-PCR) has attracted considerable interest in different research fields since it allows parallel DNA amplification on the surface of a solid substrate. However, the applications of SP-PCR have been hampered by the low efficiency of the solid-phase amplification. In order to incr...... diagnosis, high-throughput DNA sequencing, and single-nucleotide polymorphism analysis. Graphical abstract Schematic representation of solid-phase PCR....

  7. Comparison of allele-specific PCR, created restriction-site PCR, and PCR with primer-introduced restriction analysis methods used for screening complex vertebral malformation carriers in Holstein cattle

    Science.gov (United States)

    Altınel, Ahmet

    2017-01-01

    Complex vertebral malformation (CVM) is an inherited, autosomal recessive disorder of Holstein cattle. The aim of this study was to compare sensitivity, specificity, positive and negative predictive values, accuracy, and rapidity of allele-specific polymerase chain reaction (AS-PCR), created restriction-site PCR (CRS-PCR), and PCR with primer-introduced restriction analysis (PCR-PIRA), three methods used in identification of CVM carriers in a Holstein cattle population. In order to screen for the G>T mutation in the solute carrier family 35 member A3 (SLC35A3) gene, DNA sequencing as the gold standard method was used. The prevalence of carriers and the mutant allele frequency were 3.2% and 0.016, respectively, among Holstein cattle in the Thrace region of Turkey. Among the three methods, the fastest but least accurate was AS-PCR. Although the rapidity of CRS-PCR and PCR-PIRA were nearly equal, the accuracy of PCR-PIRA was higher than that of CRS-PCR. Therefore, among the three methods, PCR-PIRA appears to be the most efficacious for screening of mutant alleles when identifying CVM carriers in a Holstein cattle population. PMID:28927256

  8. Two-temperature LATE-PCR endpoint genotyping

    Directory of Open Access Journals (Sweden)

    Reis Arthur H

    2006-12-01

    Full Text Available Abstract Background In conventional PCR, total amplicon yield becomes independent of starting template number as amplification reaches plateau and varies significantly among replicate reactions. This paper describes a strategy for reconfiguring PCR so that the signal intensity of a single fluorescent detection probe after PCR thermal cycling reflects genomic composition. The resulting method corrects for product yield variations among replicate amplification reactions, permits resolution of homozygous and heterozygous genotypes based on endpoint fluorescence signal intensities, and readily identifies imbalanced allele ratios equivalent to those arising from gene/chromosomal duplications. Furthermore, the use of only a single colored probe for genotyping enhances the multiplex detection capacity of the assay. Results Two-Temperature LATE-PCR endpoint genotyping combines Linear-After-The-Exponential (LATE-PCR (an advanced form of asymmetric PCR that efficiently generates single-stranded DNA and mismatch-tolerant probes capable of detecting allele-specific targets at high temperature and total single-stranded amplicons at a lower temperature in the same reaction. The method is demonstrated here for genotyping single-nucleotide alleles of the human HEXA gene responsible for Tay-Sachs disease and for genotyping SNP alleles near the human p53 tumor suppressor gene. In each case, the final probe signals were normalized against total single-stranded DNA generated in the same reaction. Normalization reduces the coefficient of variation among replicates from 17.22% to as little as 2.78% and permits endpoint genotyping with >99.7% accuracy. These assays are robust because they are consistent over a wide range of input DNA concentrations and give the same results regardless of how many cycles of linear amplification have elapsed. The method is also sufficiently powerful to distinguish between samples with a 1:1 ratio of two alleles from samples comprised of

  9. PCR em tempo real para diagnóstico da leucose enzoótica bovina Enzootic bovine leukosis real time PCR

    Directory of Open Access Journals (Sweden)

    Natanael Lamas Dias

    2012-08-01

    Full Text Available O objetivo deste trabalho foi realizar a validação de uma reação em cadeia da polimerase em tempo real com o sistema Plexor® (qPCR para o diagnóstico da Leucose Enzoótica Bovina (LEB, por meio da comparação com testes de diagnóstico recomendados pela Organização Mundial de Saúde Animal (OIE. A qPCR foi comparada com duas outras técnicas: a PCR nested (nPCR e a imunodifusão em gel de ágar (IDGA. Das 82 amostras analisadas pela qPCR e nPCR, 79 apresentaram resultados concordantes, sendo a concordância, classificada pelo Índice Kappa, como alta. Entre as PCRs e a IDGA, o número de resultados concordantes foi de 71 e 69, respectivamente, para qPCR e nPCR, sendo a concordância classificada como considerável. A qPCR apresentou altos valores de sensibilidade e especificidade. Os valores preditivos da qPCR observados demonstraram a alta capacidade de classificação dos casos positivos e negativos. A qPCR não foi capaz de detectar três amostras positivas e tem custo ligeiramente superior que a nPCR. Entretanto, a qPCR é uma técnica mais rápida, menos susceptível a contaminações, tem alta sensibilidade, não utiliza e não gera resíduos carcinogênicos. Concluímos que a qPCR pode substituir a nPCR recomendada pela OIE no diagnóstico de rotina em áreas em que a LEB é endêmica, como no Brasil.The goal of this research was to validate a Plexor® real time Polymerase Chain Reaction (qPCR for Enzootic Bovine Leukosis (EBL diagnosis by comparison with methods recommend by the World Animal Health Organization (OIE. The qPCR was compared with two other techniques: the nested PCR (nPCR and to the agar gel immunodiffusion (AGID. Of 82 qPCR and nPCR analysed samples, 79 presented concordant results, being the concordance classified by Kappa Index as high. Between the PCRs and AGID, the number of concordant results was 71 and 69, out of 82, to qPCR and nPCR, respectively, being the concordance classified as considerable, in both

  10. Applicability of integrated cell culture reverse transcriptase quantitative PCR (ICC-RTqPCR) for the simultaneous detection of the four human enteric enterovirus species in disinfection studies

    Science.gov (United States)

    A newly developed integrated cell culture reverse transcriptase quantitative PCR (ICC-RTqPCR) method and its applicability in UV disinfection studies is described. This method utilizes a singular cell culture system coupled with four RTqPCR assays to detect infectious serotypes t...

  11. Comparison of COBAS AMPLICOR Neissefia gonorrhoeae PCR, including confirmation with N-gonorrhoeae-specific 16S rRNA PCR, with traditional culture

    NARCIS (Netherlands)

    Luijt, DS; Bos, PAJ; van Zwet, AA; Vader, PCV; Schirm, J

    A total of 3,023 clinical specimens were tested for Neisseria gonorrhoeae by using COBAS AMPLICOR (CA) PCR and confirmation of positives by N. gonorrhoeae-specific 16S rRNA PCR. The sensitivity of CA plus 16S rRNA PCR was 98.8%, compared to 68.2% for culture. Confirmation of CA positives increased

  12. Bias in the Cq value observed with hydrolysis probe based quantitative PCR can be corrected with the estimated PCR efficiency value

    NARCIS (Netherlands)

    Tuomi, Jari Michael; Voorbraak, Frans; Jones, Douglas L.; Ruijter, Jan M.

    2010-01-01

    For real-time monitoring of PCR amplification of DNA, quantitative PCR (qPCR) assays use various fluorescent reporters. DNA binding molecules and hybridization reporters (primers and probes) only fluoresce when bound to DNA and result in the non-cumulative increase in observed fluorescence.

  13. Deteccion de Chlamydia trachomatis en muestras uretrales mediante inmunofluorescencia directa Detecção de Chlamydia trachomatis em amostras uretrais mediante imunofluorescência direta Detection of Chlamydia trachomatis in urethral samples by means of direct immunofluorescence

    Directory of Open Access Journals (Sweden)

    Myra Wilson Schuster

    1989-12-01

    Full Text Available Se estudiaron 82 pacientes con uretritis para la búsqueda de Chlamydia trachomatis mediante inmunofluorescencia directa, Neisscria gonorrhoeae, Mycoplastna y Ureaplasma mediante métodos estándar. Se encontró un 19,5% de Chlamydia trachomatis y en 11 de ellos (68,8% se encontró asociada a otras bacterias y estos pacientes presentó una secreción escasa-gelatinosa.Em 82 doentes com uretrite foi pesquisada a presença de Chlamydia trachomatis, utilizando a prova da imunofluorescência direta, e de Neisseria gonorrhoeae, Mycoplasma e Ureaplasma, utilizando os métodos padrões. Ch. trachomatis foi encontrada em 19,5% dos casos, sendo que em 11 deles (68,8% observou-se associação entre Chlamydia e as outras bactérias pesquisadas. Nesses pacientes observou-se presença de secreção uretral escassa e de aspecto gelatinoso.The presence of Chlamydia trachomatis was studied by the direct immunofluorescence test, as also was that of Neisseria gonorrhoeae, Mycoplasma and Ureaplasma by the standard methods, in 82 patients with urethral discharge. Ch. trachomatis was found in 19.5% (16 of the cases and in 11 of them (68.8% there was association with the other bacteria investigated. This eleven patients presented a scanty gelatinous discharge.

  14. Capillary-based integrated digital PCR in picoliter droplets.

    Science.gov (United States)

    Chen, Jinyu; Luo, Zhaofeng; Li, Lin; He, Jinlong; Li, Luoquan; Zhu, Jianwei; Wu, Ping; He, Liqun

    2018-01-30

    The droplet digital polymerase chain reaction (ddPCR) is becoming more and more popular in diagnostic applications in academia and industry. In commercially available ddPCR systems, after they have been made by a generator, the droplets have to be transferred manually to modules for amplification and detection. In practice, some of the droplets (∼10%) are lost during manual transfer, leading to underestimation of the targets. In addition, the droplets are also at risk of cross-contamination during transfer. By contrast, in labs, some chip-based ddPCRs have been demonstrated where droplets always run in channels. However, the droplets easily coalesce to large ones in chips due to wall wetting as well as thermal oscillation. The loss of droplets becomes serious when such ddPCRs are applied to absolutely quantify rare mutations, such as in early diagnostics in clinical research or when measuring biological diversity at the cell level. Here, we propose a capillary-based integrated ddPCR system that is used for the first time to realize absolute quantification in this way. In this system, a HPLC T-junction is used to generate droplets and a long HPLC capillary connects the generator with both a capillary-based thermocycler and a capillary-based cytometer. The performance of the system is validated by absolute quantification of a gene specific to lung cancer (LunX). The results show that this system has very good linearity (0.9988) at concentrations ranging from NTC to 2.4 × 10 -4 copies per μL. As compared to qPCR, the all-in-one scheme is superior both in terms of the detection limit and the smaller fold changes measurement. The system of ddPCR might provide a powerful approach for clinical or academic applications where rare events are mostly considered.

  15. A Ribeiroia spp. (Class: Trematoda) - Specific PCR-based diagnostic

    Science.gov (United States)

    Reinitz, David M.; Yoshino, T.P.; Cole, Rebecca A.

    2007-01-01

    Increased reporting of amphibian malformations in North America has been noted with concern in light of reports that amphibian numbers and species are declining worldwide. Ribeiroia ondatrae has been shown to cause a variety of types of malformations in amphibians. However, little is known about the prevalence of R. ondatrae in North America. To aid in conducting field studies of Ribeiroia spp., we have developed a polymerase chain reaction (PCR)-based diagnostic. Herein, we describe the development of an accurate, rapid, simple, and cost-effective diagnostic for detection of Ribeiroia spp. infection in snails (Planorbella trivolvis). Candidate oligonucleotide primers for PCR were designed via DNA sequence analyses of multiple ribosomal internal transcribed spacer-2 regions from Ribeiroia spp. and Echinostoma spp. Comparison of consensus sequences determined from both genera identified areas of sequence potentially unique to Ribeiroia spp. The PCR reliably produced a diagnostic 290-base pair (bp) product in the presence of a wide concentration range of snail or frog DNA. Sensitivity was examined with DNA extracted from single R. ondatrae cercaria. The single-tube PCR could routinely detect less than 1 cercariae equivalent, because DNA isolated from a single cercaria could be diluted at least 1:50 and still yield a positive result via gel electrophoresis. An even more sensitive nested PCR also was developed that routinely detected 100 fg of the 290-bp fragment. The assay did not detect furcocercous cercariae of certain Schistosomatidae, Echinostoma sp., or Sphaeridiotrema globulus nor adults of Clinostomum sp. or Cyathocotyle bushiensis. Field testing of 137 P. trivolvis identified 3 positives with no overt environmental cross-reactivity, and results concurred with microscopic examinations in all cases. ?? American Society of Parasitologists 2007.

  16. Estudio del crecimiento de Prioria copaifera (Caesalpinaceae mediante técnicas dendrocronológicas

    Directory of Open Access Journals (Sweden)

    Jorge Andrés Giraldo Jiménez

    2011-12-01

    Full Text Available El cativo (Prioria copaifera G. forma bosques dominados por la especie llamados cativales. Durante más de 70 años el cativo ha sido la base principal de la industria maderera en el Darién colombiano, siendo, por su alta productividad y dominancia uno de los bosques tropicales más fáciles de ordenar sosteniblemente. El propósito de esta investigación es modelar el crecimiento del diámetro y el volumen del cativo en función de la edad así como las tasas instantáneas, medias y relativas, empleando anillos de crecimiento. La anualidad de los anillos se demostró usando el efecto de las bombas nucleares y mediante cofechado. Con base en los anillos de crecimiento radiales se modelo el crecimiento del diámetro y del volumen. Esta especie alcanza 40cm de diámetro en 90 años y presenta crecimiento medio de 0.31cm/año. El lapso vital de la especie es de aproximadamente 614 años. El incremento corriente anual máximo del volumen a los 90 años es 0.033m3/año, e iguala al incremento medio a los 145 años con 0.018m3/año. La tasa media de crecimiento absoluta es 0.021m3/año. Los resultados aquí obtenidos sirven como base técnica para el manejo sostenible de los cativales.A growth study of Prioria copaifera (Caesalpinaceae using dendrochronological techniques. The Cativo (Prioria copaifera forms very homogeneous forests called cativales in the flooded plains of some rivers from Costa Rica to Colombia. For over 70 years Cativo has been the main base of the timber industry in the Colombian Darien area. Because of high productivity and high-dominance of Cativo trees, they represent one of the most prone tropical forests for sustainable forest management. The objective of this research is to model diameter and timber volume growth and growth rates (absolute, mean and relative of Cativo as a function of age, using tree ring data derived from dendrochronologycal techniques. We evaluated the annual nature of the tree rings by radiocarbon

  17. Tribología de recubrimientos Cermet/NiCrBSi depositados mediante HVOF

    Directory of Open Access Journals (Sweden)

    Guilemany, J. M.

    2004-04-01

    Full Text Available This work consists on a deep tribological study of the WX system composed by a mechanical blend in different compositions of NiCrBSi and WC-12Co powders: 20% NiCrBSi (W2, 40% NiCrBSi (W4 y 60% NiCrBSi (W6. The coatings have been obtained by high velocity oxy-fuel process (HVOF. The measurements made by Ball-On-Disk test are: the friction coefficient is lower than 0.5 and the exchanged energy between the counterparts is under 10 KJ. To quantify the friction wear rate, the volume loss and the track depth, Scanning White Light Interferometry and SEM have been used. The track depth is proportional to the amount of NiCrBSi. A higher percentage of WC-12Co increases the friction wear resistance and decreases the abrasion wear rate (Rubber Wheel test. In all the coatings studied, no diffusion processes are found between the mixed phases, the adhesion between the coatings and the substrate is excellent, the porosity level is below 2% and an increase of microhardness of the coating due to a strengthening of the matrix produced by impacts of solid particles, takes place.

    El trabajo consta de un completo estudio tribológico del sistema WX que consiste en la mezcla mecánica de WC-12Co y NiCrBSi en diferentes proporciones: 20% NiCrBSi (W2, 40% NiCrBSi (W4 y 60% NiCrBSi (W6. Los recubrimientos se han obtenido por proyección térmica de alta velocidad (HVOF. Para todos los sistemas, el coeficiente de fricción es menor que 0.5 y la energía intercambiada entre el par friccionante y el recubrimiento es inferior a 10 KJ. Para cuantificar el desgaste por fricción se han utilizado la profundidad de la huella y el volumen perdido durante el ensayo, obtenidos mediante Interferometría de Barrido de Luz Blanca (SLWI y SEM. La profundidad de la huella sigue una evolución directamente proporcional al contenido en NiCrBSi. Un mayor contenido en WC-12Co aumenta la resistencia al desgaste por fricción y disminuye la velocidad de desgaste por abrasión (ensayo

  18. Falácia naturalista e naturalismo moral: do é ao deve mediante o quero

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    Adriano Naves de Brito

    2010-06-01

    Full Text Available O objetivo deste texto é discutir alguns argumentos contra a aceitação da falácia naturalista. Pretende-se mostrar aspectos que parecem corretos na argumentação dos que recusam aquela falácia, e, a despeito de seus acertos, demonstrar que estão fundamentalmente errados. O eixo para este desfecho é a reformulação da falácia em termos da recusa da implicação entre o ser e o querer. Espera-se, assim, tornar mais claras as relações - e a ausência delas - entre verdade e moral, bem como abrir espaço para a defesa de um naturalismo moral não comprometido com a existência de fatos morais. Este texto está dividido em cinco partes. Na primeira, apresenta-se a falácia naturalista de acordo com Moore; na segunda, distingue-se essa entre as formulações de Moore e de Hume; na terceira, apresentam-se objeções à interdição da passagem do "é" ao "deve" e se formulam respostas a essas objeções; na quarta, reapresenta-se o problema da interdição entre "é" e "deve" e se focaliza uma solução alternativa a ele mediante o querer; e, finalmente, na quinta parte, elaboram-se algumas conclusões acerca do naturalismo e do realismo na moral.This article aims to discuss some arguments against the acceptance of the naturalistic fallacy. It intends to demonstrate that some aspects, which seem to be correct in the argumentation of those who refuse that fallacy, are, despite their pertinence in some points, fundamentally imprecise. In order to reach this outcome, the fallacy is reformulated in terms of the refuse of the implication between "being" and "the will". Therefore, it pretends to clarify the relations - and their absence - between truth and morality as well as enable a defense of a moral naturalism disengaged from the existence of moral facts. This text is divided in five parts. In the first one, I show the naturalistic fallacy according to Moore; in the second part, a distinction between Moore's and Hume's formulations of the

  19. Application of droplet digital PCR for quantitative detection of Spiroplasma citri in comparison with real time PCR

    Science.gov (United States)

    Droplet digital Polymerase chain reaction (ddPCR) is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. It is a promising DNA quantification technology for medical diagnostics but there are only a few reports of its use for plant pat...

  20. Absolute quantification of olive oil DNA by droplet digital-PCR (ddPCR): Comparison of isolation and amplification methodologies.

    Science.gov (United States)

    Scollo, Francesco; Egea, Leticia A; Gentile, Alessandra; La Malfa, Stefano; Dorado, Gabriel; Hernandez, Pilar

    2016-12-15

    Olive oil is considered a premium product for its nutritional value and health benefits, and the ability to define its origin and varietal composition is a key step towards ensuring the traceability of the product. However, isolating the DNA from such a matrix is a difficult task. In this study, the quality and quantity of olive oil DNA, isolated using four different DNA isolation protocols, was evaluated using the qRT-PCR and ddPCR techniques. The results indicate that CTAB-based extraction methods were the best for unfiltered oil, while Nucleo Spin-based extraction protocols showed greater overall reproducibility. The use of both qRT-PCR and ddPCR led to the absolute quantification of the DNA copy number. The results clearly demonstrate the importance of the choice of DNA-isolation protocol, which should take into consideration the qualitative aspects of DNA and the evaluation of the amplified DNA copy number. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Modificación de la personalidad mediante una terapia cognitivo-conductual de afrontamiento al estrés

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    Julia Linares-Ortiz

    2014-01-01

    Full Text Available El objetivo de este estudio ha sido comprobar la posibilidad de modulación de variables de personalidad (tales como algunos de los Cinco Grandes Factores, o la personalidad resistente a través de la aplicación de un programa de afrontamiento del estrés. Para ello, han participado 26 personas del ámbito universitario con alto estrés percibido, distribuidas en dos grupos (grupo de tratamiento y grupo de control. Los instrumentos de evaluación seleccionados se clasificaron en dos grupos: variables psicológicas y emocionales y variables de personalidad. Los resultados encontrados mediantes un ANOVA de medidas repetidas mostraron que existía interacción momento x grupo en las variables optimismo, extraversión y responsabilidad, correspondientes al Modelo de los Cinco Grandes Factores, mostrando un incremento de las puntuaciones en estas variables en el grupo terapia y no encontrándose dicha interacción en el grupo control. Las puntuaciones en los componentes de neuroticismo, amabilidad, apertura a la experiencia y personalidad resistente no se modificaron en ninguno de los dos grupos. Este estudio apoya la idea de que modificando determinados parámetros emocionales relacionados con el estrés mediante terapia cognitivo-conductual se pueden ver modulados algunos factores de personalidad.

  2. DETECCIÓN DEL DISCO ÓPTICO EN RETINOGRAFÍAS MEDIANTE UNA ESTRATEGIA EVOLUTIVA (µ+λ

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    Germán Sánchez Torres

    Full Text Available En este artículo se presenta un procedimiento para la detección del disco óptico (DO en retinografías, mediante un algoritmo evolutivo. El procedimiento tiene dos etapas principales: la detección gruesa de la posición del DO y el refinamiento de los bordes del contorno. La detección gruesa ubica la posición del DO mediante un algoritmo evolutivo, cuyos individuos tienen como función objetivo la cantidad de píxeles brillantes y el número de bordes de la red de conductos sanguíneos, contenidos dentro de una circunferencia. La etapa de refinamiento aplica un procedimiento geométrico, para deformar el círculo inicial, ajustando el borde de éste con la posición del píxel de mayor variación en dirección al vector normal. El procedimiento fue evaluado empleando los repositorios públicos STARE y DIAREDB, procesando imágenes de pacientes sanos y con alteraciones de las retina, generadas por la presencia de retinopatía diabética. Los resultados experimentales muestran que el método propuesto puede identificar la posición del disco óptico en retinografías con una precisión cercana al 96 %.

  3. Detección Molecular de Toxinas Termoestable y Termolabil de Escherichia coli mediante Hibridación

    Directory of Open Access Journals (Sweden)

    Isabel Arias B

    2002-10-01

    Full Text Available Objetivos: Identificar mediante el método de hibridización por colony blot las toxinas de Escherichia coli enterotoxigénica y relacionar los resultados con los serotipos encontrados. Materiales y métodos: se evaluaron todas las cepas de E. Coli recolectadas en el Hospital de Emergencias Pediátricas de Lima durante los meses de diciembre de 1998 - abril 1999. Se usaron dos sendas de ADN que identificaban el gen de la toxina termolábil (LT y el de la toxina termoestable (ST Para la detección de los serotipos se usaron 22 antisueros de diferentes categorías de E. coli. Resultados: se encontraron 233 cepas de E. coli, 27,9% de E. coli poseían el gen LT, 3,0% el gen ST y 1,3% tenían ambos. Conclusiones: los serotipos y la presencia de los genes LT y ST no necesariamente tienen relación, demostrándose que la identificación serológica es importante en el estudio epidemiológico de diarreas causadas por E. coli debiéndose confirmar la identificación de las categorías patogénicas mediante la detección de factores de virulencia.

  4. EL APRENDIZAJE DE LOS CONCEPTOS DE FUERZAS INTERMOLECULARES E INTRAMOLECULARES MEDIANTE LA MODELIZACIÓN DIDÁCTICA

    Directory of Open Access Journals (Sweden)

    CHRISTIAN FERNNEY GIRALDO MACÍAS

    2013-07-01

    Full Text Available La modelización está siendo usada para la enseñanza y el aprendizaje en las Ciencias Naturales en diferentes contextos y para atender a diferentes problemáticas. En este caso será utilizada para explorar y analizar la relevancia que puede tener su uso en el aprendizaje de los conceptos Fuerzas Intramoleculares e Intermolecualres, partiendo de la aplicación de una serie de actividades basadas en el Ciclo Didáctico (Jorba y Sanmartí, 1996. Los datos se discuten mediante tres aspectos principales: las ideas previas de los estudiantes, el trabajo con nuevo material (nuevos conceptos, experimentos sencillos y uso de herramientas informáticas y los argumentos finales, mediante el uso de situaciones problemas. Los resultados muestran, como los estudiantes (14 y 16 años de edad evidencian un progreso conceptual al argumentar con mayor claridad las situaciones problema abordadas en el transcurso del trabajo y en la construcción de modelos más cercanos al campo científico.

  5. Manejo sostenible y sustentable de fincas productoras mediante procesos participativos en Sáchica, Boyacá

    Directory of Open Access Journals (Sweden)

    Ángel Eduardo Ramírez-Amaya

    2013-07-01

    Full Text Available Objetivo. Elaborar un proyecto de desarrollo sostenible y sustentable de fincas productoras mediante procesos participativos en el municipio de Sáchica, Boyacá. Materiales y métodos. La investigación se realizó con familias campesinas de la vereda Arrayán Alto, del municipio de Sáchica, Boyacá, mediante la metodología Investigación Acción Participativa (IAP, que se centra en la participación de las comunidades para elaborar propuestas concertadas con ellas. El trabajo se desarrolló en varias fases, que incluyeron un diagnóstico socioeconómico de las familias, capacitaciones y concientización en temas relacionados con la agricultura ecológica y de granjas integrales. Resultados. Se elaboró un plan de trabajo que permitió la construcción de un documento final que ha servido para el apoyo logístico o económico de las entidades gubernamentales locales para la instalación y plantación técnica del cultivo de gulupa con familias de la vereda Arrayán Alto.

  6. Producción de biodiesel de aceite crudo de palma mediante catálisis heterogénea

    Directory of Open Access Journals (Sweden)

    Fernando Cardeño

    2010-01-01

    Full Text Available Se estudió la producción de biodiesel de aceite crudo de palma mediante el uso de catalizadores heterogéneos ácidos y básicos para las etapas de preesterificación y transesterificación respectivamente. La pre-esterificación es necesaria para aceites con una acidez superior al 4% de ácidos grasos libres, ya que se evitan los problemas asociados con la formación de jabones. En ambas reacciones las variables analizadas fueron el tipo de catalizador, la temperatura y el tiempo. Se analizó la conversión del aceite a metilésteres usando cromatografía gaseosa y la estabilidad de los catalizadores mediante su reutilización. En la etapa de pre-esterificación se encontró que catalizadores ácidos del tipo resinas de poliestireno sulfonadas presentan conversiones superiores al 87% estabilidad para la esterificación de los ácidos grasos libres. Para la transesterificación se estudiaron como catalizadores heterogéneos, el carbonato de potasio libre y soportado en una matriz polimérica, obteniéndose con ambos conversiones superiores a 95 % de biodiesel; con el uso del soporte polimérico se disminuyó la velocidad de disolución de carbonato de potasio, permitiendo su reutilización hasta 10 reutilizaciones.

  7. Valoración de las aguas residuales mediante procedimientos analíticos y biológicos

    Directory of Open Access Journals (Sweden)

    M. Carballo

    2002-06-01

    Full Text Available Ciertos procedimientos, basados en aproximaciones analíticas y biológicas, están demostrando ser útiles en la valoración del riesgo de las aguas residuales urbanas procedentes de las Plantas de Tratamiento. Estos efluentes, considerados “mezclas complejas”, compuestos por sustancias de muy diferente naturaleza, origen y características toxicológicas y medio ambientales, requieren una valoración realista. Con el fin de colaborar al conocimiento de una parte de la realidad de nuestro país, presentamos un estudio sobre once depuradoras urbanas en las que se ha realizado un perfil de compuestos orgánicos y una valoración toxicológica mediante tests de toxicidad agudos, crónicos, de estrogenicidad, mutagenicidad y teratogenia. Los resultados muestran que 7 efluentes presentan toxicidad aguda, 3 toxicidad crónica y 4 estrogenicidad. Destacamos el hecho de que los 4 efluentes que presentan estrogenicidad, poseen al menos 3 de las sustancias estrogénicas detectadas mediante el perfil cromatográfico. Este tipo de consideraciones nos hace reflexionar sobre la necesidad de incorporar este tipo de metodologías para disponer de un conocimiento más realista de estas situaciones.

  8. Rapid diagnosis and identification by PCR of Yersinia ruckeri isolated of Oncorhynchus mykiss from Canta, Lima, Peru Diagnóstico e identificación rápidos por PCR de Yersinia ruckeri aislada de Oncorhynchus mykiss procedentes de Canta, Lima, Perú

    Directory of Open Access Journals (Sweden)

    Susana Sirvas-Cornejo

    2012-02-01

    Full Text Available Twenty individuals of rainbow trout were sampled (fry and juveniles from Acochinchan Fishfarm (Canta, Lima - Peru, and analyzed with the Polimerase Chain Reaction test (PCR in order to achieve a rapid identification of Yersinia ruckeri, which is the pathogen agent that causes the enteric red mouth disease (ERM and produces high rates of mortality. Nine fish samples were asymptomatic, while 11 of them showed signs of ERM. In addition, 22 bacterial strains were isolated from the liver, spleen and kidney. PCR and specific primers (16S rRNA, were used to amplified a specific 575 bp DNA fragment of Yersinia ruckeri. Nineteen strains were identified as Yersinia ruckeri by PCR in symptomatic and asymptomatic fishes. It was established a diagnosis time of 26 hours, compared with the 2 or 3 days that would take the diagnosis using biochemical tests.Se muestrearon 20 ejemplares (alevines y juveniles de trucha arco iris cultivados en la piscifactoría Acochinchán (Canta, Lima, Perú, y se les aplico la técnica de la Reacción en Cadena de la Polimerasa (PCR con la finalidad de obtener una identificación rápida del agente patógeno Yersinia ruckeri que produce la enfermedad entérica de la boca roja (ERM y genera elevadas tasas de mortalidad. Nueve ejemplares fueron asintomáticos mientras que 11 presentaron signos de ERM. Se aislaron 22 cepas bacterianas del hígado, bazo y riñón. Se empleó la técnica de la PCR para la amplificación y cebadores específicos (ARNr 16S, que permitieron amplificar un fragmento de ADN de 575 pb de Yersinia ruckeri. Diecinueve cepas fueron identificadas como Yersinia ruckeri mediante la PCR, tanto en peces sintomáticos como asintomáticos. Se estableció un tiempo de diagnóstico de 26 horas, en comparación con los 2 ó 3 días que duraría el diagnóstico empleando las pruebas bioquímicas.

  9. Measurement of Epstein-Barr virus DNA loads in whole blood and plasma by TaqMan PCR and in peripheral blood lymphocytes by competitive PCR.

    Science.gov (United States)

    Wadowsky, Robert M; Laus, Stella; Green, Michael; Webber, Steven A; Rowe, David

    2003-11-01

    Epstein-Barr virus (EBV) DNA load values were measured in samples of whole blood (n = 60) and plasma (n = 59) by TaqMan PCR and in samples of peripheral blood lymphocytes (PBLs) (n = 60) by competitive PCR (cPCR). The samples were obtained from 44 transplant recipients. The whole-blood and PBL loads correlated highly (r(2) > 0.900), whereas the plasma and PBL loads correlated poorly (r(2) = 0.512). Testing of whole blood by TaqMan PCR is an acceptable alternative to testing of PBLs by cPCR for quantifying EBV DNA load.

  10. Molecular methods (digital PCR and real-time PCR) for the quantification of low copy DNA of Phytophthora nicotianae in environmental samples.

    Science.gov (United States)

    Blaya, Josefa; Lloret, Eva; Santísima-Trinidad, Ana B; Ros, Margarita; Pascual, Jose A

    2016-04-01

    Currently, real-time polymerase chain reaction (qPCR) is the technique most often used to quantify pathogen presence. Digital PCR (dPCR) is a new technique with the potential to have a substantial impact on plant pathology research owing to its reproducibility, sensitivity and low susceptibility to inhibitors. In this study, we evaluated the feasibility of using dPCR and qPCR to quantify Phytophthora nicotianae in several background matrices, including host tissues (stems and roots) and soil samples. In spite of the low dynamic range of dPCR (3 logs compared with 7 logs for qPCR), this technique proved to have very high precision applicable at very low copy numbers. The dPCR was able to detect accurately the pathogen in all type of samples in a broad concentration range. Moreover, dPCR seems to be less susceptible to inhibitors than qPCR in plant samples. Linear regression analysis showed a high correlation between the results obtained with the two techniques in soil, stem and root samples, with R(2) = 0.873, 0.999 and 0.995 respectively. These results suggest that dPCR is a promising alternative for quantifying soil-borne pathogens in environmental samples, even in early stages of the disease. © 2015 Society of Chemical Industry.

  11. Ana?lisis cinemático de la marcha en pacientes con pie zambo tratados mediante el me?todo de Ponseti frente a la te?cnica quiru?rgica de liberacio?n posterior

    OpenAIRE

    Ferrando, A.; Salom Taverner, M.; Page, A.

    2018-01-01

    El objetivo principal de este proyecto consiste en valorar la evolución de la marcha en niños en edad preadolescente tratados mediante el método de Ponseti frente a los tratados mediante liberación posterior a partir de técnicas de valoración de la marcha mediante análisis biomecánico. Material y Métodos Estudio retrospectivo de casos y controles aprobado por el comité de ética. Grupo 1: 28 niños (39 pies) tratados mediante liberación posterior. Grupo 2: 18 pacientes (31 pies) tratados median...

  12. Diagnóstico genético del polimorfismo en citocromo P450 mediante ensayo de PCR múltiplex (Práctica en laboratorio virtual Cibertorio)

    OpenAIRE

    Herráez, Angel

    2017-01-01

    Guión de trabajo para el laboratorio virtual «Cibertorio» en Biomodel.UAH.es Se habla de diversidad genética de los individuos como el resultado de que, en diversas regiones del genoma, unas personas tenemos secuencias de nucleótidos que nos diferencian de otras; esto se denomina polimorfismo genético. Cuando estas regiones polimórficas corresponden a zonas codificantes de un producto, existen consecuencias funcionales. La superfamilia enzimática citocromo P450 juega un papel crucial ...

  13. El virus del papiloma humano (VPH: Un problema importante para la salud pública en vías de solución The virus of papiloma human (HPV: an important problem for the public health on the way to solution

    Directory of Open Access Journals (Sweden)

    Juan Miguel Martínez-Galiano

    2007-11-01

    Full Text Available El virus del papiloma humano es el causante de numerosos tipos de cáncer como el cáncer de cuello uterino, cáncer de pene y cáncer orofaríngeo entre otros; este virus también es el causante de lesiones como las verrugas genitales y los papilomas. Actualmente en España, la prevención del cáncer de cuello uterino se hace mediante cribado (prevención secundaria. En el año 2006 dos vacunas, una bivalente y otra tetravalente fueron aprobadas por las autoridades; estas vacunas han demostrado su seguridad y eficacia en los estudios realizados. En nuestro país, aún esta pendiente de aprobar por el Ministerio de Sanidad y Consumo pero no es así en muchos países europeos y EEUU donde ya está implantada. Hay colectivos que son resistentes a que se produzca la vacunación, la información proporcionada por los sanitarios pueden vencer esta resistencia.The virus of papiloma human is the cause of numerous types of cancer like the cancer of cervix, cancer of penis and orofaringeo cancer among others; this virus also is the cause of injuries like genital warts and papilomas. At the moment in Spain, the prevention of the cancer of cervix becomes by means of sifting (secondary prevention. In the year 2006 two vaccines, bivalent and a other tetravalente were approved by the authorities; these vaccines have demonstrated to their security and effectiveness in the made studies. In our country, still this slope to approve by the Ministry of Health and Consumption but is not thus in many European countries and the U.S.A. where already this implanted. There are groups that are resistant to that the vaccination takes place, the information provided by the sanitary personnel can overcome this resistance.

  14. Sporulation properties and antimicrobial susceptibility in endemic and rare Clostridium difficile PCR ribotypes.

    Science.gov (United States)

    Zidaric, Valerija; Rupnik, Maja

    2016-06-01

    Increased sporulation and antibiotic resistance have been proposed to be associated with certain Clostridium difficile epidemic strains such as PCR ribotype 027. In this study we examined these properties in another widespread PCR ribotype, 014/020, in comparison to prevalent PCR ribotype 002 and a group of rarely represented PCR ribotypes. Highest sporulation was observed in 014/020 strains at 24 h, while after 72 h PCR ribotype 002 and rare PCR ribotypes formed higher total number of spores. PCR ribotype 014/020 strains exhibited slightly higher resistance to tested antimicrobials, followed by group of rare PCR ribotypes and less common PCR ribotype 002. Neither sporulation properties nor antibiotic resistance clearly differed in endemic and rare strains. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. A multicenter study of viable PCR using propidium monoazide to detect Legionella in water samples.

    Science.gov (United States)

    Scaturro, Maria; Fontana, Stefano; Dell'eva, Italo; Helfer, Fabrizia; Marchio, Michele; Stefanetti, Maria Vittoria; Cavallaro, Mario; Miglietta, Marilena; Montagna, Maria Teresa; De Giglio, Osvalda; Cuna, Teresa; Chetti, Leonarda; Sabattini, Maria Antonietta Bucci; Carlotti, Michela; Viggiani, Mariagabriella; Stenico, Alberta; Romanin, Elisa; Bonanni, Emma; Ottaviano, Claudio; Franzin, Laura; Avanzini, Claudio; Demarie, Valerio; Corbella, Marta; Cambieri, Patrizia; Marone, Piero; Rota, Maria Cristina; Bella, Antonino; Ricci, Maria Luisa

    2016-07-01

    Legionella quantification in environmental samples is overestimated by qPCR. Combination with a viable dye, such as Propidium monoazide (PMA), could make qPCR (named then vPCR) very reliable. In this multicentre study 717 artificial water samples, spiked with fixed concentrations of Legionella and interfering bacterial flora, were analysed by qPCR, vPCR and culture and data were compared by statistical analysis. A heat-treatment at 55 °C for 10 minutes was also performed to obtain viable and not-viable bacteria. When data of vPCR were compared with those of culture and qPCR, statistical analysis showed significant differences (P 0.05). Overall this study provided a good experimental reproducibility of vPCR but also highlighted limits of PMA in the discriminating capability of dead and live bacteria, making vPCR not completely reliable. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Fluorescence quantitative PCR in detection of HBV DNA

    International Nuclear Information System (INIS)

    Shen Zheng; Li Ming; Shen Xia

    2003-01-01

    Objective: To study the relationship between the serum content of HBV-DNA and expression of serological markers with HBV infection patients. Methods: Serum samples from 375 hepatitis B patients with different clinical status and 70 normal persons were quantitated for HBV-DNA by FQ-PCR. Results: The average of HBV-DNA contents in the patient in the groups of HBsAg (+) and of HBeAg(+) were significantly higher than those in the group of HBsAg(-) and of HBeAg(-). Even in the group of HBeAg negative, high HBV-DNA contents might still be present in both the HBeAg(+) and HBeAg(-) groups. Conclusion: FQ-PCR can be used to monitor the real state of HBV infection, replication and the course of disease

  17. Identification of Tilletia species using rep-PCR fingerprinting technique

    Directory of Open Access Journals (Sweden)

    Župunski Vesna

    2011-01-01

    Full Text Available Analyzing 167 non-processed seed samples of wheat, it was found that 145 samples (86.8 % were contaminated with Tilletia species, while 22 (13.2 % samples were not contaminated. By using rep-PCR fingerprinting technique, it was found that DNA isolates of T. tritici originated from Serbian wheat samples had 80 % similarity with positive control for T. tritici. One isolate shared similarity of 60% with T. tritici, T. controversa and T. laevis. It was supposed that this isolate belongs to T. bromi. Isolate of T. laevis shared a similarity of 70 % with isolates of T. tritici and T. controversa, while T. walkeri was more than 10 % similar with T. tritici, T. controversa and T. laevis. Although T. controversa and T. tritici had high percent of genetic similarity, they were clustered separately. Our results suggest that rep-PCR fingerprinting could be a useful tool for monitoring presence of morphologically similar Tilletia species in wheat production areas.

  18. WetLab-2: Providing Quantitative PCR Capabilities on ISS

    Science.gov (United States)

    Parra, Macarena; Jung, Jimmy Kar Chuen; Almeida, Eduardo; Boone, Travis David; Schonfeld, Julie; Tran, Luan Hoang

    2015-01-01

    The objective of NASA Ames Research Centers WetLab-2 Project is to place on the ISS a system capable of conducting gene expression analysis via quantitative real-time PCR (qRT-PCR) of biological specimens sampled or cultured on orbit. The WetLab-2 system is capable of processing sample types ranging from microbial cultures to animal tissues dissected on-orbit. The project has developed a RNA preparation module that can lyse cells and extract RNA of sufficient quality and quantity for use as templates in qRT-PCR reactions. Our protocol has the advantage that it uses non-toxic chemicals, alcohols or other organics. The resulting RNA is transferred into a pipette and then dispensed into reaction tubes that contain all lyophilized reagents needed to perform qRT-PCR reactions. These reaction tubes are mounted on rotors to centrifuge the liquid to the reaction window of the tube using a cordless drill. System operations require simple and limited crew actions including syringe pushes, valve turns and pipette dispenses. The resulting process takes less than 30 min to have tubes ready for loading into the qRT-PCR unit.The project has selected a Commercial-Off-The-Shelf (COTS) qRT-PCR unit, the Cepheid SmartCycler, that will fly in its COTS configuration. The SmartCycler has a number of advantages including modular design (16 independent PCR modules), low power consumption, rapid thermal ramp times and four-color detection. The ability to detect up to four fluorescent channels will enable multiplex assays that can be used to normalize for RNA concentration and integrity, and to study multiple genes of interest in each module. The WetLab-2 system will have the capability to downlink data from the ISS to the ground after a completed run and to uplink new programs. The ability to conduct qRT-PCR on-orbit eliminates the confounding effects on gene expression of reentry stresses and shock acting on live cells and organisms or the concern of RNA degradation of fixed samples. The

  19. Targeted resequencing and variant validation using pxlence PCR assays

    Directory of Open Access Journals (Sweden)

    Frauke Coppieters

    2016-01-01

    Full Text Available The advent of next-generation sequencing technologies had a profound impact on molecular diagnostics. PCR is a popular method for target enrichment of disease gene panels. Using our proprietary primer-design pipeline, primerXL, we have created almost one million assays covering over 98% of the human exome. Here we describe the assay specification and both in silico and wet-lab validation of a selected set of 2294 assays using both next-generation sequencing and Sanger sequencing. Using a universal PCR protocol without optimization, these assays result in high coverage uniformity and limited non-specific coverage. In addition, data indicates a positive correlation between the predictive in silico specificity score and the amount of assay non-specific coverage.

  20. Automated 5 ' nuclease PCR assay for identification of Salmonella enterica

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Ahrens, Peter; Rådström, P.

    2000-01-01

    -point fluorescence (FAM) signals for the samples and positive control (TET) signals (relative sensitivity [Delta Rn], >0.6). The diagnostic specificity of the method was assessed using 120 non-Salmonella strains, which all resulted in negative FAM signals (Delta Rn, less than or equal to 0.5). All 100 rough...... Salmonella strains tested resulted in positive FAM and TET signals. In addition, it was found that the complete PCR mixture, predispensed in microwell plates, could be stored for up to 3 months at -20 degrees C, Thus, the diagnostic TaqMan assay developed can be a useful and simple alternative method......A simple and ready-to-go test based on a 5' nuclease (TaqMan) PCR technique was developed for identification of presumptive Salmonella enterica isolates. The results were compared with those of conventional methods. The TaqMan assay was evaluated for its ability to accurately detect 210 S. enterica...

  1. Diseño óptimo de un disipador de calor para luminaria LED mediante moderación modelación computacional

    Directory of Open Access Journals (Sweden)

    Daniel Cahue Díaz

    2014-01-01

    Full Text Available En el presente trabajo se desarrolla una selección de materiales y simulación térmica en el diseño de disipadores de calor para sistemas de iluminación de estado sólido (SSL mejor conocidos como luminarias LEDs. Se desarrolló un modelo matemático con la capacidad de predecir el comportamiento térmico de la luminaria cuando se encuentra en operación. El modelo matemático fue resuelto mediante un software de distribución libre el cual permite resolver ecuaciones diferenciales mediante el método de elemento finito. Los resultados obtenidos en el modelo matemático planteado fueron validados con los resultados obtenidos mediante experimentación usando imágenes termográficas.

  2. Inter-laboratory analysis of selected genetically modified plant reference materials with digital PCR.

    Science.gov (United States)

    Dobnik, David; Demšar, Tina; Huber, Ingrid; Gerdes, Lars; Broeders, Sylvia; Roosens, Nancy; Debode, Frederic; Berben, Gilbert; Žel, Jana

    2018-01-01

    Digital PCR (dPCR), as a new technology in the field of genetically modified (GM) organism (GMO) testing, enables determination of absolute target copy numbers. The purpose of our study was to test the transferability of methods designed for quantitative PCR (qPCR) to dPCR and to carry out an inter-laboratory comparison of the performance of two different dPCR platforms when determining the absolute GM copy numbers and GM copy number ratio in reference materials certified for GM content in mass fraction. Overall results in terms of measured GM% were within acceptable variation limits for both tested dPCR systems. However, the determined absolute copy numbers for individual genes or events showed higher variability between laboratories in one third of the cases, most possibly due to variability in the technical work, droplet size variability, and analysis of the raw data. GMO quantification with dPCR and qPCR was comparable. As methods originally designed for qPCR performed well in dPCR systems, already validated qPCR assays can most generally be used for dPCR technology with the purpose of GMO detection. Graphical abstract The output of three different PCR-based platforms was assessed in an inter-laboratory comparison.

  3. Genetic variability of Pantaneiro horse using RAPD-PCR markers

    OpenAIRE

    Egito,Andréa Alves do; Fuck,Beatriz Helena; McManus,Concepta; Paiva,Samuel Rezende; Albuquerque,Maria do Socorro Maués; Santos,Sandra Aparecida; Abreu,Urbano Gomes Pinto de; Silva,Joaquim Augusto da; Sereno,Fabiana Tavares Pires de Souza; Mariante,Arthur da Silva

    2007-01-01

    Blood samples were collected from Pantaneiro Horses in five regions of Mato Grosso do Sul and Mato Grosso States. Arabian, Mangalarga Marchador and Thoroughbred were also included to estimate genetic distances and the existing variability among and within these breeds by RAPD-PCR (Random Amplified Polymorphic DNA - Polymerase Chain Reaction) molecular markers. From 146 primers, 13 were chosen for amplification and 44 polymorphic bands were generated. The analysis of molecular variance (AMOVA)...

  4. Multiplex PCR Assay for Identification of Human Diarrheagenic Escherichia coli

    OpenAIRE

    Toma, Claudia; Lu, Yan; Higa, Naomi; Nakasone, Noboru; Chinen, Isabel; Baschkier, Ariela; Rivas, Marta; Iwanaga, Masaaki

    2003-01-01

    A multiplex PCR assay for the identification of human diarrheagenic Escherichia coli was developed. The targets selected for each category were eae for enteropathogenic E. coli, stx for Shiga toxin-producing E. coli, elt and est for enterotoxigenic E. coli, ipaH for enteroinvasive E. coli, and aggR for enteroaggregative E. coli. This assay allowed the categorization of a diarrheagenic E. coli strain in a single reaction tube.

  5. Multiple Hotspot Mutations Scanning by Single Droplet Digital PCR.

    Science.gov (United States)

    Decraene, Charles; Silveira, Amanda B; Bidard, François-Clément; Vallée, Audrey; Michel, Marc; Melaabi, Samia; Vincent-Salomon, Anne; Saliou, Adrien; Houy, Alexandre; Milder, Maud; Lantz, Olivier; Ychou, Marc; Denis, Marc G; Pierga, Jean-Yves; Stern, Marc-Henri; Proudhon, Charlotte

    2018-02-01

    Progress in the liquid biopsy field, combined with the development of droplet digital PCR (ddPCR), has enabled noninvasive monitoring of mutations with high detection accuracy. However, current assays detect a restricted number of mutations per reaction. ddPCR is a recognized method for detecting alterations previously characterized in tumor tissues, but its use as a discovery tool when the mutation is unknown a priori remains limited. We established 2 ddPCR assays detecting all genomic alterations within KRAS exon 2 and EGFR exon 19 mutation hotspots, which are of clinical importance in colorectal and lung cancer, with use of a unique pair of TaqMan ® oligoprobes. The KRAS assay scanned for the 7 most common mutations in codons 12/13 but also all other mutations found in that region. The EGFR assay screened for all in-frame deletions of exon 19, which are frequent EGFR-activating events. The KRAS and EGFR assays were highly specific and both reached a limit of detection of <0.1% in mutant allele frequency. We further validated their performance on multiple plasma and formalin-fixed and paraffin-embedded tumor samples harboring a panel of different KRAS or EGFR mutations. This method presents the advantage of detecting a higher number of mutations with single-reaction ddPCRs while consuming a minimum of patient sample. This is particularly useful in the context of liquid biopsy because the amount of circulating tumor DNA is often low. This method should be useful as a discovery tool when the tumor tissue is unavailable or to monitor disease during therapy. © 2017 American Association for Clinical Chemistry.

  6. an overview on the application of polymerase chain reaction (pcr)

    African Journals Online (AJOL)

    DR. AMINU

    Hill New York Pp818. Chul, W.P., Jang-Hee, H., Jin-Hyeok, J. et al (2004). Detection rates of Bacteria in chronic Otitis. Media with Effusion in Children, Journal of. Korean Medical Sciences 19: 735-738. Cockerill, F.R. and Smith, T.F. (2002). Rapid-Cycle real time PCR: A revolution of clinical. Microbiology. ASM News 68:2.

  7. Validation of kinetics similarity in qPCR

    Czech Academy of Sciences Publication Activity Database

    Bar, T.; Kubista, Mikael; Tichopád, Aleš

    2012-01-01

    Roč. 40, č. 4 (2012), s. 1395-1406 ISSN 0305-1048 R&D Projects: GA ČR GAP303/10/1338; GA ČR GA301/09/1752 Institutional research plan: CEZ:AV0Z50520701 Keywords : REAL-TIME PCR * POLYMERASE-CHAIN-REACTION * SYBR-GREEN-I Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 8.278, year: 2012

  8. Mycobacterium bovis in milk samples: a preliminary investigation using PCR

    International Nuclear Information System (INIS)

    Achel, D.G.; Gyamfi, O.K.; Broni, F.; Gomda, Y.; Brown, C.A.

    2007-01-01

    PCR was used to screen milk samples (n=41) for Mycobacterium bovis. DNA samples were obtained through concentration by 50% sucrose addition and centrifugation. Sixteen (16) samples (or 39%) were positive for M. Bovis DNA and the rest 25 (or 61%) were negative. All four kraals had some samples testing positive for M. bovis; the highest being 50% (5/10) and the lowest being 13% (2/15). (au)

  9. Extraction of PCR-amplifiable genomic DNA from Bacillus anthracisspores

    Energy Technology Data Exchange (ETDEWEB)

    Torok, Tamas

    2003-05-19

    Bacterial endospore disruption and nucleic acid extractionresulting in DNA of PCR-amplifiable quality and quantity are not trivial.Responding to the needs of the Hazardous Materials Response Unit (HMRU),Laboratory Division, Federal Bureau of Investigation, protocols weredeveloped to close these gaps. Effectiveness and reproducibility of thetechniques were validated with laboratory grown pure spores of Bacillusanthracis and its close phylogenetic neighbors, and with spiked soils anddamaged samples.

  10. Polymerase Chain Reaction (Pcr) Assay to Detect Hepatitis C Virus

    International Nuclear Information System (INIS)

    Lina MR; Dadang S; Budiman Bela

    2004-01-01

    Research on the detection of hepatitis C virus in blood serum using PCR technique has been carried out. Amount of 50 blood serum from laboratory of Indonesia Red Cross (Palang Merah Indonesia = PMI) and RSCM hospital as samples, were used in this research. Lysis of virus cell and extraction of RNA virus as a preliminary treatment of the sample, was done with BOOM method using guanidine thiocyanate and diatomaceous earth, respectively. Synthesis of cDNA from RNA as an extraction product mentioned above, was carried out by means of reverse-transcriptase and RNA-se inhibitor. Amplification of cDNA was done with nested PCR technique that was performed with two times PCR processes using two pairs of oligonucleotide primers for each process. The amplified DNA was detected by agarose gel electrophoresis and ethidium bromide staining. Subsequently, the DNA was visualized with UV transilluminator. Result shows that of 50 blood serum samples, 13 serum were positive for RNA HCV that were performed with the present of specific DNA band on agarose gel. (author)

  11. SCREENING OF COMMON FLAX FAD GENES BY PCR

    Directory of Open Access Journals (Sweden)

    Veronika Štefúnová

    2013-02-01

    Full Text Available Currently, flax (Linum usitatissimum L. is an important crop from commercial and economical aspects. In the spotlight is the linseed oil as a source of α-linolenic acid. The aim of presented study was to analyse fatty acid desaturase (FAD genes in flax. Several genotypes of flax (Hohenheim, La Plata 1938, Redwing USA and Escalina were used. The primers described by Vrinten et al. (2005 were used for PCR amplification reactions. Two FAD3 genes, LuFAD3A and LuFAD3B, were identified in a genome of flax. Subsequently the nucleotide sequences between origins and genotypes of flax FAD genes were compared. Primarily were used the nucleotide sequences of FAD2 and FAD3C genes available in NCBI database. Differences were found using BLAST program in nucleotide sequences of FAD genes and the specific primers were designed to amplify a specific target sequences in a genome of flax. These primers were used in PCR amplification reactions to identification of FAD2 and FAD3C genes. The PCR products were separated by electrophoresis on agarose gel.

  12. Rapid diagnosis of aneuploidy using segmental duplication quantitative fluorescent PCR.

    Directory of Open Access Journals (Sweden)

    Xiangdong Kong

    Full Text Available The aim of this study was use a simple and rapid procedure, called segmental duplication quantitative fluorescent polymerase chain reaction (SD-QF-PCR, for the prenatal diagnosis of fetal chromosomal aneuploidies. This method is based on the co-amplification of segmental duplications located on two different chromosomes using a single pair of fluorescent primers. The PCR products of different sizes were subsequently analyzed through capillary electrophoresis, and the aneuploidies were determined based on the relative dosage between the two chromosomes. Each primer set, containing five pairs of primers, was designed to simultaneously detect aneuploidies located on chromosomes 21, 18, 13, X and Y in a single reaction. We applied these two primer sets to DNA samples isolated from individuals with trisomy 21 (n = 36; trisomy 18 (n = 6; trisomy 13 (n = 4; 45, X (n = 5; 47, XXX (n = 3; 48, XXYY (n = 2; and unaffected controls (n = 40. We evaluated the performance of this method using the karyotyping results. A correct and unambiguous diagnosis with 100% sensitivity and 100% specificity, was achieved for clinical samples examined. Thus, the present study demonstrates that SD-QF-PCR is a robust, rapid and sensitive method for the diagnosis of common aneuploidies, and these analyses can be performed in less than 4 hours for a single sample, providing a competitive alternative for routine use.

  13. Authentication of Meat Species in Sucuk by Multiplex PCR

    Directory of Open Access Journals (Sweden)

    Osman İrfan İLHAK

    2015-01-01

    Full Text Available The identification of meat species used in meat products is important by reason of economic considerations, religious factors, verification of label, and prevention of unfair-market competition. In this paper, multiplex PCR method was experienced for routine detection of equine (horse and donkey, poultry (chicken and turkey, pig and cattle meat in sucuk (sausage. The primers used for these animals generated specific fragments, and they did not show cross reactions with the DNA from the other genus of animal. After multiplex PCR was successfully optimized, a field study was carried out to investigate the presence of horse, donkey, chicken, turkey and pig meat in 50 sucuks (30 beef and 20 beef + poultry collected from markets. The result of the field study indicated that 23.3% of 30 beef sucuk samples were containing poultry meat. None of the 50 sucuk samples was containing pig meat, but one (2% of the samples generated equine fragment. The present study showed that the multiplex PCR method can be used for routine analysis of meat species identification, verification and control of label information of meat products.

  14. Direct-to-PCR tissue preservation for DNA profiling.

    Science.gov (United States)

    Sorensen, Amy; Berry, Clare; Bruce, David; Gahan, Michelle Elizabeth; Hughes-Stamm, Sheree; McNevin, Dennis

    2016-05-01

    Disaster victim identification (DVI) often occurs in remote locations with extremes of temperatures and humidities. Access to mortuary facilities and refrigeration are not always available. An effective and robust DNA sampling and preservation procedure would increase the probability of successful DNA profiling and allow faster repatriation of bodies and body parts. If the act of tissue preservation also released DNA into solution, ready for polymerase chain reaction (PCR), the DVI process could be further streamlined. In this study, we explored the possibility of obtaining DNA profiles without DNA extraction, by adding aliquots of preservative solutions surrounding fresh human muscle and decomposing human muscle and skin tissue samples directly to PCR. The preservatives consisted of two custom preparations and two proprietary solutions. The custom preparations were a salt-saturated solution of dimethyl sulfoxide (DMSO) with ethylenediaminetetraacetic (EDTA) and TENT buffer (Tris, EDTA, NaCl, Tween 20). The proprietary preservatives were DNAgard (Biomatrica(®)) and Tissue Stabilising Kit (DNA Genotek). We obtained full PowerPlex(®) 21 (Promega) and GlobalFiler(®) (Life Technologies) DNA profiles from fresh and decomposed tissue preserved at 35 °C for up to 28 days for all four preservatives. The preservative aliquots removed from the fresh muscle tissue samples had been stored at -80 °C for 4 years, indicating that long-term archival does not diminish the probability of successful DNA typing. Rather, storage at -80 °C seems to reduce PCR inhibition.

  15. Aspergillus section Fumigati typing by PCR-restriction fragment polymorphism.

    Science.gov (United States)

    Staab, Janet F; Balajee, S Arunmozhi; Marr, Kieren A

    2009-07-01

    Recent studies have shown that there are multiple clinically important members of the Aspergillus section Fumigati that are difficult to distinguish on the basis of morphological features (e.g., Aspergillus fumigatus, A. lentulus, and Neosartorya udagawae). Identification of these organisms may be clinically important, as some species vary in their susceptibilities to antifungal agents. In a prior study, we utilized multilocus sequence typing to describe A. lentulus as a species distinct from A. fumigatus. The sequence data show that the gene encoding beta-tubulin, benA, has high interspecies variability at intronic regions but is conserved among isolates of the same species. These data were used to develop a PCR-restriction fragment length polymorphism (PCR-RFLP) method that rapidly and accurately distinguishes A. fumigatus, A. lentulus, and N. udagawae, three major species within the section Fumigati that have previously been implicated in disease. Digestion of the benA amplicon with BccI generated unique banding patterns; the results were validated by screening a collection of clinical strains and by in silico analysis of the benA sequences of Aspergillus spp. deposited in the GenBank database. PCR-RFLP of benA is a simple method for the identification of clinically important, similar morphotypes of Aspergillus spp. within the section Fumigati.

  16. Aspergillus Section Fumigati Typing by PCR-Restriction Fragment Polymorphism▿

    Science.gov (United States)

    Staab, Janet F.; Balajee, S. Arunmozhi; Marr, Kieren A.

    2009-01-01

    Recent studies have shown that there are multiple clinically important members of the Aspergillus section Fumigati that are difficult to distinguish on the basis of morphological features (e.g., Aspergillus fumigatus, A. lentulus, and Neosartorya udagawae). Identification of these organisms may be clinically important, as some species vary in their susceptibilities to antifungal agents. In a prior study, we utilized multilocus sequence typing to describe A. lentulus as a species distinct from A. fumigatus. The sequence data show that the gene encoding β-tubulin, benA, has high interspecies variability at intronic regions but is conserved among isolates of the same species. These data were used to develop a PCR-restriction fragment length polymorphism (PCR-RFLP) method that rapidly and accurately distinguishes A. fumigatus, A. lentulus, and N. udagawae, three major species within the section Fumigati that have previously been implicated in disease. Digestion of the benA amplicon with BccI generated unique banding patterns; the results were validated by screening a collection of clinical strains and by in silico analysis of the benA sequences of Aspergillus spp. deposited in the GenBank database. PCR-RFLP of benA is a simple method for the identification of clinically important, similar morphotypes of Aspergillus spp. within the section Fumigati. PMID:19403766

  17. Validation of chimerism in pediatric recipients of allogeneic hematopoietic stem cell transplantation (HSCT) a comparison between two methods: real-time PCR (qPCR) vs. variable number tandem repeats PCR (VNTR PCR).

    Science.gov (United States)

    Kletzel, Morris; Huang, Wei; Olszewski, Marie; Khan, Sana

    2013-01-01

    Post-hematopoietic stem cell transplantation (HSCT) chimerism monitoring is important to assess relapse and therapeutic intervention. The purpose of our study is to compare two methods variable number tandem repeats (VNTR) vs. quantitative real- time polymerase chain reaction (qPCR) in terms of determining chimerism. 127 (peripheral blood n=112, bone marrow n=15) samples were simultaneously tested by VNTR using APO-B, D1S80, D1S111, D17S30, gene loci SRY and ZP3 and qPCR using 34 assays (CA001-CA034) that are designed to a bi-allelic insertion/deletion (indel) polymorphism in the human genome. Samples were separated in three subsets: total WBC, T-cell and Myeloid cells. Extraction of DNA was performed then quantified. We analyzed column statistics, paired t-test and regression analysis for both methods. There was complete correlation between the two methods. The simplicity and rapidity of the test results from the qPCR method is more efficient and accurate to assess chimerism.

  18. Diagnosis of aerobic vaginitis by quantitative real-time PCR.

    Science.gov (United States)

    Rumyantseva, T A; Bellen, G; Savochkina, Y A; Guschin, A E; Donders, G G G

    2016-07-01

    To evaluate a real-time PCR-based technique to quantify bacteria associated with aerobic vaginitis (AV) as a potential test. Vaginal samples from 100 women were tested by wet-mount microscopy, gram stain and quantitative real-time PCR targeting Enterobacteriacea, Staphylococcus spp., Streptococcus spp., Enterococcus spp., Escherichia coli, Streptococcus agalactiae, S. aureus; Lactobacillus spp. AV diagnosis obtained by wet-mount microscopy was used as reference. Some level of AV was diagnosed in 23 (23.7 %) cases. Various concentrations of Enterobacteriacea, Staphylococcus spp., Streptococcus spp. were detected an all patients. Enterococcus spp. were detected in 76 (78.3 %) cases. Summarized concentrations of aerobes were tenfold higher in AV-positive compared to AV-negative cases [7.30lg vs 6.06lg (p = 0.02)]. Concentrations of aerobes in severe, moderate and light AV cases did not vary significantly (p = 0.14). Concentration of lactobacilli was 1000-fold lower in AV-positive cases compared to normal cases (5.3lg vs 8.3lg, p AV-positive cases [19/22 (86.4 %) samples]. The relation of high loads of aerobes to the low numbers of Lactobacilli are a reliable marker for the presence of AV and could substitute microscopy as a test. PCR may be a good standardized substitution for AV diagnosis in settings where well-trained microscopists are lacking.

  19. Using PCR to Target Misconceptions about Gene Expression

    Directory of Open Access Journals (Sweden)

    Leslie K. Wright

    2013-02-01

    Full Text Available We present a PCR-based laboratory exercise that can be used with first- or second-year biology students to help overcome common misconceptions about gene expression. Biology students typically do not have a clear understanding of the difference between genes (DNA and gene expression (mRNA/protein and often believe that genes exist in an organism or cell only when they are expressed. This laboratory exercise allows students to carry out a PCR-based experiment designed to challenge their misunderstanding of the difference between genes and gene expression. Students first transform E. coli with an inducible GFP gene containing plasmid and observe induced and un-induced colonies. The following exercise creates cognitive dissonance when actual PCR results contradict their initial (incorrect predictions of the presence of the GFP gene in transformed cells. Field testing of this laboratory exercise resulted in learning gains on both knowledge and application questions on concepts related to genes and gene expression.

  20. Orbitotomía lateral mediante abordaje temporal Lateral orbitotomy using a temporal approach

    Directory of Open Access Journals (Sweden)

    H. Herencia Nieto

    2005-12-01

    Full Text Available La orbitotomía lateral sigue siendo en el momento actual la técnica quirúrgica de elección para la biopsia o extirpación de lesiones intraorbitarias laterales al nervio óptico, la biopsia del propio nervio óptico y la extirpación de la glándula lacrimal. Se han descrito múltiples incisiones cutáneas para llevar a cabo esta intervención; la más frecuentemente empleada por el momento es la incisión palpebral, que discurre a nivel de alguna arruga del párpado superior. Aunque los resultados obtenidos con esta incisión suelen ser aceptables, no está exenta de complicaciones, ni estéticas ni funcionales. Por esto, en los últimos años han ido apareciendo nuevas incisiones que intentan evitar estas complicaciones. Entre estas nuevas vías de abordaje a la pared lateral de la órbita se encuentra la incisión temporal. En los últimos 3 años, se ha intervenido en nuestro servicio a cuatro pacientes para llevar a cabo biopsia o extirpación de masas intraorbitarias mediante orbitotomía lateral, siempre a través de una incisión temporal. En todos los casos esta incisión permitió una amplia exposición del campo quirúrgico y la cómoda realización de la intervención, obteniéndose resultados estéticos y funcionales excelentes, y sin ninguna secuela ni complicación permanente. La complicación específica más frecuente de esta incisión es la paresia de la rama frontal del nervio facial. Una técnica de disección cuidadosa suele ser suficiente para evitarla. Todo esto hace que esta incisión, tal y como la describimos aquí, sea de elección para llevar a cabo la orbitotomía lateral, constituyendo en nuestro criterio una alternativa perfectamente válida y a tener en cuenta frente a las incisiones «clásicas», a las que llega a superar en muchos aspectos.The lateral orbitotomy it still the surgical technique of choice for biopsies or the removal of intraorbital lesions that are lateral to the optic nerve, for biopsies of

  1. Purification of nanoparticle PCR products and their topography observed with AFM

    International Nuclear Information System (INIS)

    Mi Lijuan; Wang Hubin; Chinese Academy of Sciences, Beijing; Li Bin; Zhou Hualan; Hu Jun

    2007-01-01

    Nanoparticle PCR (NP-PCR) is a new method to optimize PCR amplification. Suitable amount of Au nanoparticles can improve specificity, sensitivity and extension rate of PCR. In this paper, we compare efficiency of purifying NP-PCR products with different methods. In addition, topographies of DNA products in NP-PCR were observed with atomic force microscope (AFM). The results show that most of DNA products purified directly by routing method remain almost free due to less effect of nanoparticales. The yields decrease when the AuNPs were removed by high-speed centrifugation. A little amount of DNA subsided with AuNPs. (authors)

  2. Estudio de la recuperación de cromo hexavalente mediante un reactor electroquímico de compartimentos separados por separadores cerámicos

    OpenAIRE

    REYES PINEDA, HENRY

    2011-01-01

    La Tesis Doctoral "Estudio de la recuperación de cromo hexavalente mediante un reactor electroquímico de compartimentos separados por separadores cerámicos" se centra en la posibilidad de recuperación del cromo hexavalente procedente de las disoluciones de mordentado de las industrias de metalizado de plásticos mediante la utilización de un reactor electroquímico de compartimentos separados por separadores cerámicos fabricados a diferente presión y diferente composición de almidón. Con la rec...

  3. Atenuación de la asimetría y de la curtosis de las puntuaciones observadas mediante transformaciones de variables: Incidencia sobre la estructura factorial

    OpenAIRE

    Miguel Ángel Ruiz Díaz; María Noel Rodríguez Ayán

    2008-01-01

    En este trabajo se evalúa la incidencia de la atenuación, mediante transformaciones de variables, del sesgo y de la curtosis de las puntuaciones observadas, sobre la estructura factorial, estimada mediante análisis factorial exploratorio y confirmatorio. Los datos proceden de una escala de opinión estudiantil para la evaluación de profesores universitarios, de 16 ítems medidos en escala Likert. Las distribuciones observadas no se aproximan a la normalidad, por lo que ...

  4. Valoración nutricional mediante curvas de crecimiento de la OMS y las clasificaciones de Gómez / Waterlow. Estudio de prevalencia. Cuenca-2015

    OpenAIRE

    Chacón Abril, Karla Lorena; Segarra Ortega, José Xavier; Lasso Lazo, Rubén Santiago; Huiracocha Tutivén, María de Lourdes

    2016-01-01

    OBJETIVO:Determinar la prevalencia de malnutrición mediante las curvas de crecimiento (OMS) y de desnutrición según la clasificación Gómez/Waterlow; establecer ventajas y desventajas del empleo de ambos sistemas de clasificación.MÉTODOS:Estudio de prevalencia realizado en el Subcentro de Salud Sinincay, con una población de 737 niños/as registrados en la matriz de vigilancia alimentaria y nutricional (SIVAN) durante Enero-Junio 2015, que identificó la malnutrición infantil mediante el uso de ...

  5. Procedimiento para la discriminación y mapeo de los rodales de nerdo en cultivos de girasol mediante teledetección

    OpenAIRE

    López Granados, Francisca; García Torres, Luis; Peña Barragán, José Manuel; Jurado-Expósito, Montserrat

    2006-01-01

    Procedimiento para la discriminación y mapeo de los rodales de nerdo en cultivos de girasol mediante teledetección. Procedimiento para mapear zonas infestadas de la mala hierba conocida como nerdo (Ridolfia segetum Moris) en plantaciones de girasol mediante teledetección. Tiene aplicación en Agricultura, y más concretamente en Empresas de Asistencia Técnica Agraria o Medioambiental, o en Auditorias Agroambientales Públicas o Privadas. Principalmente consiste en el anál...

  6. Caracterización de polipropileno con fibra de vidrio y policarbonato/acrilonitrilo butadieno estireno microespumados mediante moldeo por inyección MuCell®

    OpenAIRE

    Rojas Jiménez, Ana

    2016-01-01

    El proyecto tiene como objetivo el análisis morfológico y de propiedades mecánicas de placas de polipropileno con fibra de vidrio (PP-GF30) y de policarbonato/acrilonitrilo butadieno estireno (PC/ABS), inyectadas mediante microespumación física (MuCell®). Este proyecto se enmarca dentro de un estudio más amplio que tiene como objetivo la comparación entre dos métodos de espumado físico mediante moldeo por inyección: el proceso MuCell® y un nuevo proceso del grupo Volkswagen...

  7. La participación de los trabajadores en el capital social mediante operaciones de asistencia financiera. Especial referencia al art. 81.2 LSA

    OpenAIRE

    Nieto Rojas, Patricia

    2007-01-01

    La integración de los trabajadores en el capital social puede ser instrumentada mediante diferentes vías: distribución gratuita de acciones, entrega de opciones sobre acciones o mediante la asistencia financiera para la adquisición de acciones; el objetivo del presente artículo es analizar las implicaciones laborales de este último mecanismo previsto en el artículo 81.2 LSA que exceptúa de la prohibición general de asistencia financiera a los negocios dirigidos a facilitar la adquisición de a...

  8. Análisis de la biodiversidad de macroinvertebrados bentónicos del río Cunas mediante indicadores ambientales, Junín-Perú

    OpenAIRE

    María Custodio Villanueva; Fernán Cosme Chanamé Zapata

    2016-01-01

    El objetivo de la investigación fue analizar el estado de la biodiversidad de macroinvertebrados bentónicos del río Cunas mediante indicadores ambientales. Se definieron tres sectores de muestreo en dos épocas contrastantes. La valoración de las presiones antrópicas se realizó mediante la determinación de la carga de DBO5 aportada por aguas residuales. Se colectaron muestras de agua para la determinación de nitratos, fosfatos y coliformes termotolerantes. Los indicadores medidos in situ fuero...

  9. Estudio comparativo del comportamiento de losas de concreto reforzado mediante los análisis elástico y límite

    Directory of Open Access Journals (Sweden)

    Julio Vergara García

    1989-01-01

    Full Text Available Presenta un resumen de la tesis de grado "Estudio comparativo de losas de concreto reforzado mediante los análisis elástico y límite". Este trabajo proporciona fórmulas, tablas y gráficas prácticas para determinar los momentos flectores y el volumen de refuerzo de los tipos de losas estudiados, sometidas a diferentes tipos de carga y analizados mediante la Teoría de la Elasticidad y el Análisis Límite.

  10. "DETECCIÓN DE TRASTORNO DE DEFICIT DE ATENCIÓN MEDIANTE LA ESCALA DE CONNERS EN NIÑOS 6 A 9 AÑOS DE EDAD"

    OpenAIRE

    Cancino Estrada, Yurixhi

    2012-01-01

    Antecedentes: el TDAH es la enfermedad psiquiátrica crónica más frecuente en la edad pediátrica. Su prevalencia es del 3 al 5%. No existe una encuesta única para sospechar TDAH, una de ellas es la escala de Conners, al detectar oportunamente a estos niños e iniciar tratamiento, se disminuye fracaso escolar y rechazo social. Objetivo: detectar TDAH mediante la escala de Conners en niños de 6 a 9 años de edad. Material y métodos: estudio descriptivo mediante aplicación de l...

  11. Diferencias en la calciuria, estimada mediante el índice Ca/Cr en función del tipo de lactancia

    OpenAIRE

    Trigo López, Javier

    2015-01-01

    Se ha realizado un estudio descriptivo transversal sobre el comportamiento de la calciuria, estimada mediante el índice calcio/creatinina (ICC), en lactantes menores de 6 meses, analizando su posible relación con el tipo de alimentación (leche de fórmula o leche materna). Los resultados se obtuvieron a partir de muestras de orina de 44 lactantes sanos en los que se recoge el tipo de lactancia. El grupo alimentado mediante leche de fórmula presentó un ICC medio expresado en mg/mg de 0,59, mien...

  12. Obtención de extractos de membrana externa de Vibrio cholerae O1, mediante el uso de diferentes detergentes

    Directory of Open Access Journals (Sweden)

    José Luis Pérez

    2006-04-01

    Full Text Available En la actualidad existen dos variantes principales de vacunas orales contra el cólera: una basada en células inactivadas de diferentes biotipos y serotipos y otra basada en la administración de cepas vivas genéticamente atenuadas. Una vacuna por subunidades pudiera ser una variante muy atractiva. Este trabajo describe la purificación parcial y caracterización preliminar de extractos de proteínas de membrana externa-lipopolisacárido (PME-LPS, obtenidos a partir de Vibrio cholerae O1, con el interés de seleccionar un proteoliposoma que posteriormente será estructurado en forma de cocleatos para su uso por vía oral en humanos. Las preparaciones fueron obtenidas a través del uso de diferentes detergentes. La cantidad de LPS en cada preparación fue estimada mediante la determinación de las unidades endotóxicas en el ensayo del Limulus (LAL. La composición de cada muestra fue evaluada mediante SDS-PAGE y Dot Blot. La inoculación intranasal (IN en ratones Balb/c se utilizó para la evaluación de la inmunogenicidad de las preparaciones, y la respuesta inmune fue determinada por ELISA y el título de anticuerpos vibriocidas. El tamaño molecular de la preparación con mejores resultados en inmunogenicidad se estimó mediante la cromatografía en Sephacryl S-1000. Se obtuvieron diferentes perfiles electroforéticos de acuerdo con el tipo de detergente utilizado. El LPS fue identificado en todas las preparaciones y aquella obtenida con el SDS al 15% mostró la más baja relación proteínas/LPS y los mejores resultados en los ensayos de inmunogenicidad. Adicionalmente se comprobó que su tamaño molecular es similar al observado en el proteoliposoma de VAMENGOC- BC. La preparación obtenida con el SDS al 15% constituye un proteoliposoma, con capacidad para estimular altos niveles de anticuerpos IgG anti-LPS y altos títulos de anticuerpos vibriocidas, luego de su administración por vía intranasal en ratones. Estos resultados constituyen

  13. Reconocimiento del habla mediante el uso de la correlación cruzada y una perceptrón multicapa

    Directory of Open Access Journals (Sweden)

    Carlos A. de Luna-Ortega

    2014-01-01

    Full Text Available En el presente artículo se da a conocer una alternativa algorítimica a los sistemas actuales de reconocimiento automático del habla (ASR, mediante una propuesta en la forma de realizar la caracterización de las palabras basada en una aproximación que usa la extracción de coeficientes de la codificación de predicción lineal (LPC y la correlación cruzada. La implementación consiste en extraer las características fonéticas mediante los coeficientes LPC, después se forman vectores de patrones de la pronunciación conformados por el promedio de los coeficientes LPC de las muestras de las palabras obteniendo un vector característico de cada pronunciación mediante la autocorrelación de las secuencias de coeficientes LPC; estos vectores se utilizan para entrenar un clasificador de tipo perceptrón multicapa (MLP. Se realizaron pruebas de desempeño previo entrenamiento con los diferentes patrones de las palabras a reconocer. Se utilizó la fonética de los dígitos del cero al nueve como vocabulario objetivo, debido a su amplia aplicación, y para estimar el desempeño de este método se utilizaron dos corpus de pronunciaciones: el corpus UPA, que contempla en su base de datos la pronuncación de la región occidente de México, y el corpus Tlatoa, que hace lo propio para la región centro de México. Las señales en ambos corpus fueron adquiridas en el lenguaje español, y a una frecuencia de muestreo de 8kHz. Los porcentajes de reconocimiento obtenidos fueron del 96.7 y 93.3% para las modalidades de mono-locutor para el corpus UPA y múltiple-locutor para el corpus Tlatoa, respectivamente. Asimismo, se realizó una comparación contra dos métodos clásicos del reconocimiento de voz y del habla, Dynamic Time Warping (DTW y Hidden Markov Models (HMM.

  14. Vida a la fatiga de juntas soldadas del acero inoxidable AISI 316L obtenidas mediante el proceso GMAW

    Directory of Open Access Journals (Sweden)

    Puchi-Cabrera, E. S.

    2007-06-01

    Full Text Available An investigation has been conducted in order to determine the effect of both the metallic transfer mode (pulsed arc or short circuit and the O2 content in the Ar/O2 gas mixture, of the gas-metal arc welding process (GMAW, on the fatigue life under uniaxial conditions of welded joints of 316L stainless steel. It has been concluded that the mixture of the protective gases employed in the process could have an important influence on the fatigue life of the welded joints of such steel in two different ways. Firstly, through the modification of the radius of curvature at the joint between the welding toe and the base metal and, secondly, through a more pronounced degree of oxidation of the alloying elements induced by a higher O2 content in the mixture. As far as the metallic transfer mode is concerned, it has been determined that the welded joints obtained under a pulsed arc mode show a greater fatigue life in comparison with the joints obtained under short circuit for both gas mixtures.

    Se ha llevado a cabo una investigación con la finalidad de determinar el efecto, tanto del modo de transferencia metálica (arco pulsado o cortocircuito como del contenido de O2 en la mezcla de gases protectores Ar/O2, del proceso de soldadura a tope mediante arco metálico con protección gaseosa (GMAW, sobre la vida a la fatiga en condiciones uniaxiales de juntas soldadas del acero inoxidable AISI 316L. Dicho trabajo ha permitido concluir que la composición de la mezcla de gases protectores del proceso GMAW pudiera tener una influencia importante en la vida a la fatiga de las juntas soldadas de dicho material, a través de dos formas distintas: primero, mediante la modificación del radio de curvatura entre la raíz del cordón de soldadura y el metal base y, en segundo lugar, a través del mayor grado de oxidación de los elementos de aleación. En cuanto al modo de transferencia metálica, se determinó que las juntas soldadas mediante arco pulsado

  15. A new detection method for the K variant of butyrylcholinesterase based on PCR primer introduced restriction analysis (PCR-PIRA).

    Science.gov (United States)

    Shibuta, K; Abe, M; Suzuki, T

    1994-01-01

    The K variant of human butyrylcholinesterase is caused by a G/A transition in the butyrylcholinesterase gene, which neither creates nor destroys any restriction site. In an attempt to detect the K variant both simply and rapidly, we developed a two step method of "PCR primer introduced restriction analysis" (PCR-PIRA). The first step was used to introduce a new Fun4HI site into the normal allele for a screening test, while the second step was performed to create a new MaeIII site on the variant allele for a specific test. This method thus enabled us to distinguish clearly the K variant from the normal allele, and also showed that the frequency of the K variant allele is 0.164 in the Japanese population. Images PMID:7966197

  16. Implementation of polymerase chain reaction (PCR and Real-Time PCR in quick identification of bovine herpesvirus 1

    Directory of Open Access Journals (Sweden)

    Milić Nenad

    2010-01-01

    Full Text Available Examinations were performed on 65 samples of nasal smeas taken from calves and young cows with clinical symptoms of respiratory infection to determine the presence of the bovine herpes virus 1 using parallel implementation of molecular and standard methods of virological diagnostics. The appearance of a cytopathogenic effect (CPE was not established in inoculated cell lines 24h, 48h and 72h following inoculation, or after two successive passages of the examined material sample through these cell lines. The application of polymerize chain reaction (PCR using a primer for glucoprotein B and thymidine - kinasis, established the presence of bovine herpes virus 1 nucleic acid in one sample of a bovine nasal smear, while the presence of this virus was established in three samples in an examination of the nasal smear samples using the Real-Time PCR method.

  17. A comparison of QuantStudio™ 3D Digital PCR and ARMS-PCR for measuring plasma EGFR T790M mutations of NSCLC patients.

    Science.gov (United States)

    Feng, Qin; Gai, Fei; Sang, Yaxiong; Zhang, Jie; Wang, Ping; Wang, Yue; Liu, Bing; Lin, Dongmei; Yu, Yang; Fang, Jian

    2018-01-01

    The AURA3 clinical trial has shown that advanced non-small cell lung cancer (NSCLC) patients with EGFR T790M mutations in circulating tumor DNA (ctDNA) could benefit from osimertinib. The aim of this study was to assess the usefulness of QuantStudio™ 3D Digital PCR System platform for the detection of plasma EGFR T790M mutations in NSCLC patients, and compare the performances of 3D Digital PCR and ARMS-PCR. A total of 119 Chinese patients were enrolled in this study. Mutant allele frequency of plasma EGFR T790M was detected by 3D Digital PCR, then 25 selected samples were verified by ARMS-PCR and four of them were verified by next generation sequencing (NGS). In total, 52.94% (69/119) had EGFR T790M mutations detected by 3D Digital PCR. In 69 positive samples, the median mutant allele frequency (AF) was 1.09% and three cases presented low concentration (AF Digital PCR) was identified as T790M- by ARMS-PCR. Four samples were identified as T790M+ by both NGS and 3D Digital PCR, and typically three samples (3/4) presented at a low ratio (AF Digital PCR is a novel method with high sensitivity and specificity to detect EGFR T790M mutation in plasma.

  18. Development of Nested PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Cylindrocladium scoparium on Eucalyptus

    Directory of Open Access Journals (Sweden)

    Tian-Min Qiao

    2016-10-01

    Full Text Available Eucalyptus dieback disease, caused by Cylindrocladium scoparium, has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP were developed for detection of C. scoparium based on factor 1-alpha (tef1 and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium. The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products.

  19. Development of Nested PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Cylindrocladium scoparium on Eucalyptus

    Science.gov (United States)

    Qiao, Tian-Min; Zhang, Jing; Li, Shu-Jiang; Han, Shan; Zhu, Tian-Hui

    2016-01-01

    Eucalyptus dieback disease, caused by Cylindrocladium scoparium, has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP) were developed for detection of C. scoparium based on factor 1-alpha (tef1) and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium. The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products. PMID:27721691

  20. Development of Nested PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Cylindrocladium scoparium on Eucalyptus.

    Science.gov (United States)

    Qiao, Tian-Min; Zhang, Jing; Li, Shu-Jiang; Han, Shan; Zhu, Tian-Hui

    2016-10-01

    Eucalyptus dieback disease, caused by Cylindrocladium scoparium , has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP) were developed for detection of C. scoparium based on factor 1-alpha (tef1) and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium . The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products.

  1. Revealing the microbiota of marketed edible insects through PCR-DGGE, metagenomic sequencing and real-time PCR.

    Science.gov (United States)

    Osimani, Andrea; Milanović, Vesna; Garofalo, Cristiana; Cardinali, Federica; Roncolini, Andrea; Sabbatini, Riccardo; De Filippis, Francesca; Ercolini, Danilo; Gabucci, Claudia; Petruzzelli, Annalisa; Tonucci, Franco; Clementi, Francesca; Aquilanti, Lucia

    2018-07-02

    The present study aimed to identify the microbiota present in six species of processed edible insects produced in Thailand and marketed worldwide via the internet, namely, giant water bugs (Belostoma lutarium), black ants (Polyrhachis), winged termites (alates, Termitoidae), rhino beetles (Hyboschema contractum), mole crickets (Gryllotalpidae), and silkworm pupae (Bombyx mori). For each species, two samples of boiled, dried and salted insects were purchased. The microbial DNA was extracted from the insect samples and subjected to polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), high-throughput sequencing and qualitative real-time PCR assays. The microbiota of the analyzed samples were widely characterized by the presence of spore-forming bacteria mainly represented by the genera Bacillus and Clostridium. Moreover, the genera Anaerobacillus, Paenibacillus, Geobacillus, Pseudomonas, Stenotrophomonas, Massilia, Delftia, Lactobacillus, Staphylococcus, Streptococcus, Vagococcus, and Vibrio were also detected. Real-time PCR allowed for ascertainment of the absence of Coxiella burnetii, Shiga toxin-producing E. coli (STEC), and Pseudomonas aeruginosa in all samples. The results of this study confirm the importance of combining different molecular techniques to characterize the biodiversity of complex ecosystems such as edible insects. The presence of potential human pathogens suggests the need for a careful application of good manufacturing practices during insect processing. This study provides further data that will be useful in risk analyses of edible insects as a novel food source. Copyright © 2018 Elsevier B.V. All rights reserved.

  2. Highly efficient PCR assay to discriminate allelic DNA methylation status using whole genome amplification

    Directory of Open Access Journals (Sweden)

    Ito Takashi

    2011-06-01

    Full Text Available Abstract Background We previously developed a simple method termed HpaII-McrBC PCR (HM-PCR to discriminate allelic methylation status of the genomic sites of interest, and successfully applied it to a comprehensive analysis of CpG islands (CGIs on human chromosome 21q. However, HM-PCR requires 200 ng of genomic DNA to examine one target site, thereby precluding its application to such samples that are limited in quantity. Findings We developed HpaII-McrBC whole-genome-amplification PCR (HM-WGA-PCR that uses whole-genome-amplified DNA as the template. HM-WGA-PCR uses only 1/100th the genomic template material required for HM-PCR. Indeed, we successfully analyzed 147 CGIs by HM-WGA-PCR using only ~300 ng of DNA, whereas previous HM-PCR study had required ~30 μg. Furthermore, we confirmed that allelic methylation status revealed by HM-WGA-PCR is identical to that by HM-PCR in every case of the 147 CGIs tested, proving high consistency between the two methods. Conclusions HM-WGA-PCR would serve as a reliable alternative to HM-PCR in the analysis of allelic methylation status when the quantity of DNA available is limited.

  3. [Sensitivity and specificity of nested PCR pyrosequencing in hepatitis B virus drug resistance gene testing].

    Science.gov (United States)

    Sun, Shumei; Zhou, Hao; Zhou, Bin; Hu, Ziyou; Hou, Jinlin; Sun, Jian

    2012-05-01

    To evaluate the sensitivity and specificity of nested PCR combined with pyrosequencing in the detection of HBV drug-resistance gene. RtM204I (ATT) mutant and rtM204 (ATG) nonmutant plasmids mixed at different ratios were detected for mutations using nested-PCR combined with pyrosequencing, and the results were compared with those by conventional PCR pyrosequencing to analyze the linearity and consistency of the two methods. Clinical specimens with different viral loads were examined for drug-resistant mutations using nested PCR pyrosequencing and nested PCR combined with dideoxy sequencing (Sanger) for comparison of the detection sensitivity and specificity. The fitting curves demonstrated good linearity of both conventional PCR pyrosequencing and nested PCR pyrosequencing (R(2)>0.99, PNested PCR showed a better consistency with the predicted value than conventional PCR, and was superior to conventional PCR for detection of samples containing 90% mutant plasmid. In the detection of clinical specimens, Sanger sequencing had a significantly lower sensitivity than nested PCR pyrosequencing (92% vs 100%, Pnested PCR and Sanger sequencing method, nested PCR pyrosequencing has a higher sensitivity especially in clinical specimens with low viral copies, which can be important for early detection of HBV mutant strains and hence more effective clinical management.

  4. Módulo de identificación biométrica mediante huellas dactilares para sistemas empotrados

    OpenAIRE

    Juste Meco, Rubén

    2010-01-01

    La seguridad es uno de los temas que más preocupan en la actualidad, pero no solo en lo referente a la integridad física del individuo, sino también en lo concerniente a la protección de sus bienes o datos. En la sociedad actual se hace uso de multitud de servicios electrónicos para disfrutar de los bienes, e-banking, compras mediante el uso de tarjetas de crédito, e-mail, e-shopping; y dispositivos electrónicos para acceder a los datos, PCs, PDAs, Teléfonos Móviles y demás. Sin una seguridad...

  5. SISTEMA DE ASEGURAMIENTO DE LA CALIDAD MEDIANTE LA APLICACIÓN DEL SISTEMA HACCP EN LA INDUSTRIA DE PASTAS ALIMENTICIAS

    Directory of Open Access Journals (Sweden)

    Aleida González González

    2005-09-01

    Full Text Available

    En este trabajo se presenta un estudio del HACCP (Hazard Analysis Critical Control Points como un sistema de aseguramiento de la calidad en lo referente a la inocuidad para la industria de alimentos, mencionando las técnicas generalmente usadas para llevar a acabo este estudio y mostrando un ejemplo mediante su aplicación en la industria de pastas alimenticias. Como resultado se elaboró la tabla de control HACCP, la misma que recopila los PCC, el peligro posible, los límites críticos, las medidas correctivas, sistema de monitoreo y responsables de las acciones tomadas.

  6. Eliminación de cromo de efluentes ácidos, mediante adsorción con wollastonita natural

    OpenAIRE

    Martín Antonio Encinas Romero; Luis Alberto Núñez Rodríguez; Agustín Gómez Álvarez; Guillermo Del Carmen Tiburcio Munive

    2015-01-01

    El presente trabajo evalúa las características de la remoción de cromo con wollastonita natural a partir de soluciones sintéticas de cromo (VI) en medio ácido. Para realizar este estudio se desarrolló un diseño factorial 23 en el cual se estudiaron los factores: relación sólido/líquido, concentración de cromo en solución y temperatura. Se analizó el efecto de los factores principales y sus interacciones sobre el porcentaje de remoción de cromo mediante análisis gráficos. Asimismo, se desarr...

  7. Determinación de un polimorfo de la cimetidina mediante la calorimetría diferencial de barrido

    Directory of Open Access Journals (Sweden)

    Luis Martínez Álvarez

    1997-12-01

    Full Text Available Se estableció la diferencia entre 2 sustancias de masa molecular similar -la cimetidina A y la B-, pero de estructuras internas distintas, mediante el empleo de la técnica de calorimetría diferencial de barrido donde solamente una de ellas, la cimetidina B, posee la actividad farmacológica requerida, es decir, cumple con las especificaciones de la farmacopea.The difference between two substances of similar mollecular mass but of different internal structuras -cimetidine A and B- was established by using the differential scanning calorimetry. It was proved that only one of them, cimetidine B, has the required pharmacological activity, which means that it fulfills the pharmacopoeia specifications.

  8. Generación de experiencias visuales en ciegos mediante estimulación táctil repetitiva

    Directory of Open Access Journals (Sweden)

    Tomás Ortiz

    2012-02-01

    Full Text Available Estudios recientes de nuestro laboratorio han establecido que se puede generar una activación estable del córtex visual en ciegos mediante entrenamiento táctil pasivo prolongado. Esta neuroplasticidad cortical se acompaña en un 40% de los participantes invidentes de sensaciones visuales, así como de una mayor rapidez en el reconocimiento táctil de información espacial tales como líneas, iconos o imágenes, y es crucial para su correcta interpretación. Estos hallazgos pueden ser útiles para el diseño de mecanismos sustitutivosde la visión en ciegos, pero también son útiles para entender ciertos síntomas neuropsiquiátricos como lassinestesias.

  9. SIMULACIÓN MEDIANTE REDES NEURONALES DEL ENFRIAMIENTO DEL ARCO ELÉCTRICO EN DISYUNTORES DE ALTA TENSIÓN

    Directory of Open Access Journals (Sweden)

    ALEXANDRO BRYANTSEVICH

    2010-07-01

    Full Text Available La formulación de los modelos analíticos de enfriamiento de los arcos eléctricos en disyuntores de alta tensión todavía presenta muchas dificultades, y requiere nuevas hipótesis que los simplifique sustancialmente en relación con la realidad. Desde un punto de vista técnico, la aplicación de las redes neuronales podría ser una fuente objetiva para la simulación del enfriamiento del arco eléctrico, y para obtener resultados más cercanos a las propiedades reales del interruptor. En este trabajo se describe la problemática del desarrollo de modelos analíticos vinculados al enfriamiento del arco eléctrico de disyuntores de alta tensión mediante redes neuronales.

  10. Procedimiento para aumentar la velocidad de obtención de biodiesel mediante su incorporación como aditivo

    OpenAIRE

    López Granados, Manuel; Mariscal López, Rafael; Martín Alonso, David; Bretes García, Pilar

    2008-01-01

    Procedimiento para aumentar la velocidad de obtención de biodiésel mediante su incorporación como aditivo. La presente invención se refiere al uso de biodiésel como aditivo para aumentar la velocidad de reacción en reacciones de transesterificación catalítica con alcoholes y para proteger a los catalizadores del contacto con el CO{sub,2} y H{sub,2}O atmosféricos. De forma más concreta, en esta invención se describe un procedimiento de obtención de biodiésel a partir de la transesterificación ...

  11. Marcador de brecha mediante fotomicrografía sobre fibroblastos cultivados en Petri para determinar el efecto de la electroterapia

    OpenAIRE

    Naranjo Fraile, Aida

    2017-01-01

    En este proyecto se pretende crear una aplicación biomédica con la que se analicen de forma automática imágenes micrográficas para la obtención de resultados sobre una investigación. Ésta se basa en el cierre de brecha en dermis humana mediante la aplicación de terapia eléctrica. Las imágenes son obtenidas en el Servicio de Bioelectromagnetismo del Hospital Universitario Ramón y Cajal de Madrid. La interfaz ha sido creada con el programa MatLab, en concreto con el entorno de programación de i...

  12. REGULACIÓN ECONÓMICA PARA LA TARIFA DE PARQUEADEROS EN BOGOTÁ MEDIANTE PRECIOS MÁXIMOS

    Directory of Open Access Journals (Sweden)

    Jorge Andrés Perdomo Calvo

    2012-12-01

    Full Text Available El objetivo principal de este estudio consiste en estimar el valor máximo o techo mediante el cual deberían estar reguladas las tarifas del servicio de parqueadero en Bogotá, a partir de los lineamientos teóricos de la economía del transporte y bajo el criterio de fijación de precios. Los resultados de este análisis se evidencian con base en información primaria (encuestas, modelos de costo total de producción no lineales (Box-Cox y optimización matemática (estática comparativa. Se concluye que el precio por minuto actual se encuentra sobrevalorado.

  13. Medición de contaminación mediante UAV (Vehículo Aéreo no Tripulado

    Directory of Open Access Journals (Sweden)

    Edwin José Vera-Rozo

    2016-09-01

    Full Text Available Este artículo presenta un procedimiento experimental cuyo objetivo es obtener la medición de contaminación en un relleno sanitario (basurero mediante un Vehículo Aéreo no Tripulado (UAV; La metodología utilizada consistió en realizar un procedimiento detallado para la instrumentación de UAV, el cual fue equipamiento con un sistema para la captura y almacenamiento de datos referente a las variables medidas en tiempo real, el cual posteriormente se puso en vuelo y luego se realizo el procesamiento de la información offline; para finalizar se presentan los resultados obtenidos y conclusiones.

  14. Detection of Mycobacterium tuberculosis in extrapulmonary biopsy samples using PCR targeting IS6110, rpoB, and nested-rpoB PCR Cloning.

    Science.gov (United States)

    Meghdadi, Hossein; Khosravi, Azar D; Ghadiri, Ata A; Sina, Amir H; Alami, Ameneh

    2015-01-01

    Present study was aimed to examine the diagnostic utility of polymerase chain reaction (PCR) and nested PCR techniques for the detection of Mycobacterium tuberculosis (MTB) DNA in samples from patients with extra pulmonary tuberculosis (EPTB). In total 80 formalin-fixed, paraffin-embedded (FFPE) samples comprising 70 samples with definite diagnosis of EPTB and 10 samples from known non- EPTB on the basis of histopathology examination, were included in the study. PCR amplification targeting IS6110, rpoB gene and nested PCR targeting the rpoB gene were performed on the extracted DNAs from 80 FFPE samples. The strong positive samples were directly sequenced. For negative samples and those with weak band in nested-rpoB PCR, TA cloning was performed by cloning the products into the plasmid vector with subsequent sequencing. The 95% confidence intervals (CI) for the estimates of sensitivity and specificity were calculated for each method. Fourteen (20%), 34 (48.6%), and 60 (85.7%) of the 70 positive samples confirmed by histopathology, were positive by rpoB-PCR, IS6110-PCR, and nested-rpoB PCR, respectively. By performing TA cloning on samples that yielded weak (n = 8) or negative results (n = 10) in the PCR methods, we were able to improve their quality for later sequencing. All samples with weak band and 7 out of 10 negative samples, showed strong positive results after cloning. So nested-rpoB PCR cloning revealed positivity in 67 out of 70 confirmed samples (95.7%). The sensitivity of these combination methods was calculated as 95.7% in comparison with histopathology examination. The CI for sensitivity of the PCR methods were calculated as 11.39-31.27% for rpoB-PCR, 36.44-60.83% for IS6110- PCR, 75.29-92.93% for nested-rpoB PCR, and 87.98-99.11% for nested-rpoB PCR cloning. The 10 true EPTB negative samples by histopathology, were negative by all tested methods including cloning and were used to calculate the specificity of the applied methods. The CI for 100

  15. Control de bacteriemia nosocomial pediátrica mediante un programa de cultivo de soluciones parenterales en uso

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    Muñoz Juan M.

    1999-01-01

    Full Text Available OBJETIVO. Dado que Klebsiella, Enterobacter y Serratia se multiplican en soluciones parenterales y son responsables de una elevada proporción de bacteriemias en los hospitales de México, se propone una estrategia de control mediante la vigilancia microbiológica de las soluciones en uso. MATERIAL Y MÉTODOS. Hospital de enseñanza de segundo nivel con 193 camas. Atiende principalmente pacientes de escasos recursos. En 1992 se inició la vigilancia de la esterilidad de las soluciones parenterales en los servicios pediátricos mediante cuatro estrategias: durante la primera etapa se cultivó el total de soluciones en uso. Durante la segunda se cultivaron muestras aleatoriamente elegidas. Tercera y cuarta etapas con muestreo controlado y dirigido, respectivamente. RESULTADOS. Se han cultivado 1940 infusiones. Se ha observado una reducción de la tasa de contaminación (de 29.6% en 1992 a 12.9% en 1997, p< 0.001. Asimismo se redujo la proporción de bacilos gramnegativos aislados en sangre (72.7% vs 40.85%, p< 0.001 y las bacteriemias nosocomiales primarias (BNP (3.12 vs 1.54 por 100 egresos, p< 0.0001. CONCLUSIONES. La detección de contaminantes señala posibles fallas en el manejo parenteral, áreas de riesgo y pacientes potencialmente afectados. El programa permite estudiar el nivel endémico de contaminación de infusiones y limitar los brotes de bacteriemias nosocomiales primarias a un costo bajo.

  16. Cálculo del poder de la lente intraocular mediante biometría ultrasónica

    Directory of Open Access Journals (Sweden)

    Yileika Elías García

    Full Text Available Objetivo: caracterizar el comportamiento del cálculo de la lente intraocular mediante biometría ultrasónica. Métodos: se realizó un estudio descriptivo y prospectivo, con un grupo de pacientes atendidos por el Servicio de Catarata del Centro Oftalmológico «Enrique Cabrera», que recibieron tratamiento quirúrgico por catarata con implante de lente intraocular, mediante técnica de Blumenthal. La muestra se conformó con 160 ojos y se estudiaron las variables: sorpresa refractiva, edad, longitud axial y promedio queratométrico según resultado refractivo, poder dióptrico de la lente y componente esférico según edad. Resultados: como resultados más sobresalientes, se obtuvo que el 78,7 % no presentó error en el cálculo de la lente, la mayoría de los pacientes tenían 60 años y más, predominó el rango de longitud axial entre 22 y 24,4 (67,5 %, el promedio queratométrico de 43 a 44,99 (64,3 %, el poder dióptrico del lente de + 20 a + 23 dioptrías (48,2 % y el componente esférico entre 0,00 y +/-1,00 dioptrías (78,8%. Conclusiones: se considera que la biometría ultrasónica resulta adecuada para el cálculo correcto del lente.

  17. Solución al Problema de Secuenciación de Trabajos mediante el Problema del Agente Viajero

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    G.E. Anaya Fuentes

    2016-10-01

    Full Text Available Resumen: En este trabajo se estudia el Problema de Secuenciación de Trabajos codificado como un Problema de Agente Viajero y resuelto mediante Algoritmos Genéticos. Se propone un Algoritmo Genético en donde se comparan dos tipos de selección: por torneo y por ruleta. Se realizan diferentes pruebas para la solución del Problema del Agente Viajero con los dos tipos de selección bajo diferentes parámetros: número de individuos, número de iteraciones, probabilidad de cruce y probabilidad de mutación; a partir de estos se seleccionan los parámetros y el tipo de selección. Posteriormente se codifica al Problema de Secuenciación como un Problema del Agente Viajero. La propuesta se presenta mediante la aplicación a diferentes ejemplos del Problema de Secuenciación de Trabajos y la comparación con los resultados obtenidos en la literatura. Abstract: In this paper we proposed a solution to the Job-Shop Scheduling Problem using the Traveling Salesman Problem solved by Genetic Algorithms. We proposed a genetic algorithm where we compare two types of selection: tournament and roulette. Different tests are performed to solve the Traveling Salesman Problem with the two types of selection under different parameters: number of individuals, number of iterations, crossover probability and mutation probability. Then the best type of selection and the best parameters are used to solve the Job-Shop Scheduling Problem with Genetic Algorithms for the Traveling Salesman Problem. The proposal is presented solving different examples of Job Sequencing Problem and compare them with the results obtained in the literature. Palabras clave: algoritmos eficientes, sistemas industriales de producción, problemas de optimización, problema de agente viajero, Keywords: Efficient algorithms, industrial production systems, optimization problem, traveling salesman problem

  18. DISTRIBUCIÓN ESPACIAL DE LA TUBERCULOSIS EN ESPAÑA MEDIANTE MÉTODOS GEOESTADÍSTICOS

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    Diana Gómez-Barroso

    2009-01-01

    Las tasas de TB respiratoria experimentan un descenso constante en los últimos años. El objetivo es valorar la asociación entre la morbilidad por TB respiratoria y variables socioeconómicas y epidemiológicas así como su distribución espacial mediante métodos geoestadísticos. Método: Las tasas de incidencia se estandarizaron por edad y sexo con datos de la Red Nacional de Vigilancia (2006. Las variables socioeconómicas incluidas son: condición socioeconómica, nivel de estudios, tasa de hacinamiento, densidad de población, tasa de inmigración estandarizada por sexo, tasa de analfabetismo, tasa de paro, gasto medio en euros por persona. Las variables epidemiológicas incluidas han sido la tasa de SIDA y la tasa de incidencia de gripe. Se realizó un análisis multivariable mediante un Modelo Lineal Generalizado poisson. Se aplicó la técnica geoestadística Cokringing ajustada por las variables estadísticamente significativas para ver la distribución espacial de riesgo. Resultados: Las variables estadísticamente significativas son la tasa de hacinamiento, tasa de inmigración, tasa de analfabetismo, tasa de paro, gasto medio euros por persona, tasa de gripe y tasa de sida. La técnica geoestadística muestra una variabilidad espacial del riesgo y una concentración del riesgo en el noroeste y sureste de la península. Conclusiones: Los resultados permiten afirmar que el método Cokriging es una herramienta útil para representar la distribución espacial del riesgo. Existe asociación entre variables socioeconómicas, epidemiológicas y TB en España.

  19. Elaboración de aleaciones de Cu-Al-Ni con efecto memoria de forma mediante pulvimetalurgia

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    Pérez-Sáez, R. B.

    1998-05-01

    Full Text Available During the production of shape memory alloys, a very fine grain size should be obtained in order to obtain better mechanical properties and a good thermomechanical behaviour during cycling. The classically used grain refiners show some secondary effects on the martensitic transformations that could be at the origin of some technological problems. For this reason, a new processing method by powder metallurgy has been developed for this kind of alloys. The three proceeding stages are described: Atomization, hot isostatic pressing and hot rolling. The microstructure of the materials is characterized. The martensitic transformation and the thermomechanical properties are also studied.

    En la elaboración de aleaciones Cu-Al-Ni con efecto memoria de forma es importante conseguir un tamaño de grano fino, para mejorar las propiedades mecánicas y el comportamiento durante el ciclado termomecánico. Clásicamente, esto se ha conseguido mediante la adición de refinadores de grano; sin embargo, los efectos secundarios que éstos producen pueden ser problemáticos. Por esta razón, se ha desarrollado un nuevo método de procesado de este tipo de aleaciones mediante pulvimetalurgia. En este trabajo se presenta el proceso de elaboración consistente en tres etapas: atomización, compactación isostática en caliente y laminación en caliente. Se estudia la microestructura del material, se caracteriza la transformación martensítica y se determinan las propiedades termomecánicas.

  20. MULTIPLEX SYBR® GREEN-REAL TIME PCR (qPCR ASSAY FOR THE DETECTION AND DIFFERENTIATION OF Bartonella henselae AND Bartonella clarridgeiae IN CATS

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    Rodrigo Staggemeier

    2014-04-01

    Full Text Available A novel SYBR® green-real time polymerase chain reaction (qPCR was developed to detect two Bartonella species, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.