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 ...

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...

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

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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

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 ...

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.

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 ...

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.

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

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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.

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

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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

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...

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

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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

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

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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

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

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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.

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...

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

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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.

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.

Aceptabilidad de la vacuna contra el virus del papiloma humano en madres de la provincia de Valencia (España)

OpenAIRE

Navarro Illana, Pedro; Caballero Pérez, Pablo; Tuells Hernández, José; Puig Barberá, Joan; Díez-Domingo, Javier

2015-01-01

Introducción: La Comunidad Valenciana inició en octubre del 2008 el programa de vacunación contra el virus del papiloma humano (VPH) en niñas de 14 años. El objetivo de este estudio es evaluar los conocimientos sobre la infección por VPH y su vacuna en madres de adolescentes e identificar los factores asociados a la predisposición de vacunar a sus hijas. Material y métodos: Estudio observacional transversal mediante cuestionario dirigido a madres de alumnas nacidas en 1995 matriculadas en cen...

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

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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

Utilidad en la combinación de oligonucleótidos universales para la detección del virus del papiloma humano en cáncer cervicouterino y lesiones premalignas Usefulness of combining universal oligonucleotides in detecting human papillomavirus in cervical cancer and premalignant lesions

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Adela Carrillo

2004-02-01

Full Text Available OBJETIVO: Determinar la frecuencia y distribución del virus del papiloma humano en los diferentes estadios que conforman la historia natural del cáncer cérvico uterino, y optimizar la detección mediante el uso de diferentes oligonucleótidos universales. MATERIAL Y MÉTODOS: Se trata de un estudio transversal, descriptivo, en el que las muestras fueron colectadas durante enero a diciembre de 1999. El procesamiento de las muestras y el análisis de los datos se realizaron en el Instituto Nacional de Cancerología en la Ciudad de México. Se hizo análisis comparativo con t de Student para valores continuos y con ji cuadrada para proporciones, y análisis de concordancia entre biopsia y exudado cervical con la prueba estadística de Kappa. Para la detección del virus se utilizó la técnica de la reacción en cadena de la polimerasa (PCR con oligonucleótidos universales los cuales reconocen diferentes regiones del gen L1 (MY09/11; GP5/6; L1C1/2, y oligonucleótidos específicos para el VPH 16 y el VPH 18, así como secuenciación directa de los productos de la PCR. RESULTADOS: Se analizaron 154 muestras: 65 (42.2% citologías normales, 45 (29.2% lesiones de alto y bajo grado, y 44 (28.6% de cáncer invasor. El VPH fue detectado en 95.5% de los casos de cáncer invasor, en 91.6% de lesiones de alto grado, en 66.7% de lesiones de bajo grado y en 23.1% de citologías normales, por la PCR con al menos uno de los juegos de oligonucleótidos utilizados. La detección fue más eficiente en las muestras obtenidas por biopsia que en los exudados cervicovaginales. El porcentaje total de detección del VPH con un juego de oligonucleótidos universales (37.6% aumentó sustancialmente (60.4% al combinarlo con otros dos juegos de oligonucleótidos universales. CONCLUSIONES: La presencia del VPH de alto riesgo es elevada inclusive en mujeres con epitelios cervicales con diagnóstico citológico normal. La detección del VPH mejora al utilizar distintos

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

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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

[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.

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

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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

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...

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

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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.

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

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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.

LA HIPERQUERATOSIS CITOLOGICA COMO PARAMETRO INDIRECTO DE LA INFECCIÓN POR EL VIRUS DEL PAPILOMA HUMANO.

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Victoria Martín Gómez

2009-01-01

Full Text Available Introducción. La hiperqueratosis es una lesión epitelial que histológicamente se caracteriza, por presentar un aumento del grosor de la capa epitelial superficial cornificada, con ausencia de núcleos. Entre las causas que favorecen la aparición de hiperqueratosis, se puede resaltar la reacción del epitelio a los estímulos locales, mecánicos, químicos o infecciosos, siendo más relevante en estos casos el virus del papiloma humano. Objetivo: Conocer la relación que existe entre la presencia de hiperqueratosis en los estudios citológicos de cribado y la posible relación con la infección del virus del Papiloma Humano. Material y métodos: Se han estudiado un total de 2372 extendidos citológicos realizados durante un año en el Servicio de Obstetricia y Ginecología del Hospital Clínico de Salamanca. La edad de las pacientes estaba comprendida entre los 16 y 65 años. La toma citológica realizada mediante la técnica de triple toma se depositaba en medio líquido. Se descartaron las citologías que presentaban alteraciones citológicas, y se valoraron 1125 estudios citológicos considerados como negativos. De estos, 316 presentaron hiperqueratosis. A las pacientes que presentaban hiperqueratosis se les realizó estudio colposcópico y determinación de VPH por PCR. Resultados: En el 27% de las hiperqueratosis, la PCR de VPH fue negativa, el resto, positiva. Se ha podido determinar un VPP del 72% y un VPN del 39%. La sensibilidad fue del 46% y la Especificidad del 81%.

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

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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.

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...

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

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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

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

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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

Lesión de Virus Papiloma Humano a nivel del labio en paciente escolar

OpenAIRE

Millán Isea, Ronald E; Ferrer, Maria A; Pérez, Ligia

2007-01-01

RESUMEN El virus Papiloma Humano (VPH), es un virus ADN, que produce proliferación cutánea o mucosa de epitelio escamoso estratificado. Existen más de 80 subtipos de VPH, los cuales pueden ser identificados por diferentes estudios histopatológicos convencionales, a través del cual se pueden determinar los cambios celulares en los queratinocitos e inmunohistoquímicos y más especializados como hibridación in situ y la técnica de reacción en cadena de polimerasa (PCR), donde puede identificarse ...

Factores asociados a lesiones cervicales o presencia del virus del papiloma humano en dos poblaciones de estudiantes de Lima

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María Valderrama C

2007-07-01

Full Text Available Objetivos: Determinar la prevalencia y factores asociados a lesiones cervicales o presencia del virus del papiloma humano (VPH en mujeres estudiantes en educación superior de 18 a 26 años de Lima. Materiales y métodos: Se realizó un estudio de corte transversal, en dos universidades y un instituto superior tecnológico de Lima, durante los meses de agosto a diciembre del 2001. Se aplicó un cuestionario y se colectaron muestras para Papanicolaou (PAP y detección del ADN de los VPH 6, 11, 16, 18 por el método de PCR (reacción en cadena de la polimerasa. Se incluyeron en el análisis 321 estudiantes que reportaron actividad sexual a quienes se tomó muestras para PAP y VPH. Resultados: La prevalencia de VPH (6, 11, 16, 18 fue de 8,4%, y para las lesiones cervicales fue 2,5% (diagnóstico a través del PAP. Las lesiones cervicales o presencia del VPH fueron más frecuentes en el grupo de 21 a 23 años (p= 0,024. La diferencia de edades (tres a más años entre la pareja sexual de mayor edad y la participante se asoció significantemente con lesiones cervicales o presencia del VPH (OR:8,8; IC95:1,9-39,6. La edad de la primera relación sexual, número de parejas sexuales y uso de condón, no mostraron significancia estadística. Conclusiones: Las lesiones cervicales o presencia del VPH son frecuentes en esta población de mujeres jóvenes. La edad y la diferencia de edades con la pareja sexual de mayor edad se asociaron a las lesiones cervicales o presencia del VPH.

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

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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

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

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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

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

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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.

ESTRUCTURA MOLECULAR Y ANTIGÉNICA DE LA VACUNA

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VÍCTOR ANDRÉS VANEGAS

2008-09-01

Full Text Available La proteína L1 del Virus del Papiloma Humano (VPH constituye el 80% de la cápside viral. Las vacunas profilácticas contra el VPH son sintetizadas a partir de la proteína L1 ensamblada en Partículas similares al Virus (del inglés VLP, las cuales son altamente inmunogénicas generando anticuerpos específicos de tipo y en algunos casos pueden presentar reacción cruzada entre tipos de VPH filogenéticamente próximos. La estructura de la proteína L1 del VPH es importante porque confiere estabilidad a la cápside mediante el establecimiento de interacciones intra e intercapsoméricas lo que asegura la integridad viral y antigénicamente porque contiene los epítopes que inducen la respuesta inmune protectora. En estudios en los que se evaluó la antigenicidad de la proteína L1 se determinó que los epítopes inmunodominantes de la cápside viral se encuentran en los bucles B-C, D-E, F-G, H-I y en el extremo C-terminal. Estos bucles son poco conservados entre los diferentes genotipos y se encuentran en segmentos de la proteína expuestos en la superficie de la cápside. Los aminoácidos situados en los bucles B-C, F-G y H-I son primordiales para el reconocimiento por los anticuerpos neutralizantes. Los diferentes subtipos y variantes presentan cambios en estos aminoácidos o en residuos que conforman otros epítopes. En esta revisión se presentará un estado del arte de la proteína L1 del VPH genotipo 16, la estructura y su importancia en el desarrollo de vacunas contra la infección producida por este virus.

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

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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.

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.

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...

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

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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.

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

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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

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

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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...

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.

Human Papillomavirus infection in men residing in Brazil, Mexico, and the USA Infección por Virus de Papiloma Humano en hombres de Brasil, México y EUA

Directory of Open Access Journals (Sweden)

2008-10-01

Virus del Papiloma Humano (VPH entre hombres de 18 años o más de tres países con un protocolo común para el muestreo de la detección de VPH, y evaluar si la detección de VPH varía de acuerdo con la edad y el país. MATERIAL Y MÉTODOS: El estudio incluye diversas etapas que inician con la identificación de hombres susceptibles, una medición basal (visita de enrolamiento y nueve visitas adicionales programadas cada seis meses. En este artículo, se presenta el análisis de los primeros 1160 hombres que fueron incluídos en el estudio. Para maximizar la posibilidad de detección de VPH se utilizó un cepillo de dacrón que muestreó en forma combinada diferentes sitios anatómicos. Para la determinación de ADN de VPH se utilizó ión en cadena de polimerasa (PCR por amplificación de un fragmento del gen de VPH L1. RESULTADOS: Entre 1160 hombres de Brasil, México y EUA, la prevalencia global de VPH fue de 65.2%, con solamente 12% de tipos oncogénicos, 20.7% de tipos de VPH no oncogénicos, 17.8% de muestras positivas a tipos oncogénicos y no oncogénicos; y finalmente 14.7% de infecciones no clasificadas. Múltiples tipos de VPH fueron detectados en 25.7% de los participantes en el estudio. La prevalencia de VPH fue más alta en Brasil (72.3%, comparada con la observada en EUA (61.3% y México (61.9%. Los tipos de VPH 16 (6.5%, 51 (6.5% y 59 (5.3% fueron los más comúnmente observados con poder oncogénico. El VPH 84 (7.7%, 62 (7.3% y 6 (6.6% fueron las infecciones no oncogénicas más comunes. CONCLUSIONES: Son necesarios estudios de la distribución de VPH en un amplio margen de edad entre hombres de múltiples países, para establecer con mayor precisión, el conocimiento de la historia natural de la infección por VPH en hombres.

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

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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

Country-level correlates of cervical cancer mortality in Latin America and the Caribbean Determinantes a nivel país de la mortalidad por cáncer cervicouterino en Latinoamérica y el Caribe

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Ana Pereira-Scalabrino

2013-02-01

Full Text Available OBJECTIVE: To identify country-level correlates of geographical variations in cervical cancer (CC mortality in Latin America and the Caribbean (LAC. MATERIALS AND METHODS: CC mortality rates for LAC countries (n=26 were examined in relation to country-specific socio-economic indicators (n=58 and Human Papilloma Virus (HPV prevalence using linear regression models. RESULTS: High mortality at ages OBJETIVO: Identificar variables a nivel de país que expliquen las variaciones geográficas en la mortalidad por cáncer cervicouterino (CaCu en América Latina y el Caribe (AL. MATERIALES Y MÉTODOS: Se examinaron las tasas de mortalidad por CaCu de cada país (n=26 mediante modelos de regresión lineal en relación con indicadores socioeconómicos (n=58 y prevalencia del virus del papiloma humano (VPH. RESULTADOS: Alta mortalidad en menores de cinco años, bajo gasto total en salud per-cápita y baja proporción de población con acceso a saneamiento básico son los mejores predictores de mortalidad por CaCu (R² =77%. En los países (n=10 con estimaciones de prevalencia de VPH, estos indicadores socioeconómicos y la prevalencia de VPH de alto riesgo explicaron el 98% de la variabilidad de CaCu en AL. CONCLUSIÓN: Las mejoras en el nivel socioeconómico en AL están asociadas con reducciones en la mortalidad por CaCu, a pesar de la ausencia de programas organizados de tamizaje e inmunización contra VPH.

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

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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

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.

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

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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.

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

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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.

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.

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

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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

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.

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 ...

Determinantes sociales de la salud de la OMS en mujeres mexicanas con el virus de papiloma

OpenAIRE

Soltero-Rivera, Silvia Guadalupe; Cerda-Flores, Ricardo Martin; Cárdenas-Villarreal, Velia Margarita; Guevara-Valtier, Milton Carlos; Paz Morales, María de los Angeles; Patton-Leal, Adrián Carlos; Ramírez-García, Esther Justina

2016-01-01

Uno de los temas centrales de la OMS es el análisis de los problemas de salud mediante el modelo de Determinantes Sociales de la Salud (DSS: inadecuadas condiciones económicas, ambientales y de alimentación). El sistema sanitario de la OMS ha asociado algunas enfermedades tal como el Virus de Papiloma Humano (VPH) con losDSS. Desde el punto de vista de investigación cuantitativa, los DSS reportados por la OMS son: 1) edad de inicio de la actividad sexual, 2) múltiples compañeros sexuales, 3) ...

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.

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...

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.

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

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

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…

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.

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.

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...

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.

The Virtual Physiological Human ToolKit.

Science.gov (United States)

Cooper, Jonathan; Cervenansky, Frederic; De Fabritiis, Gianni; Fenner, John; Friboulet, Denis; Giorgino, Toni; Manos, Steven; Martelli, Yves; Villà-Freixa, Jordi; Zasada, Stefan; Lloyd, Sharon; McCormack, Keith; Coveney, Peter V

2010-08-28

The Virtual Physiological Human (VPH) is a major European e-Science initiative intended to support the development of patient-specific computer models and their application in personalized and predictive healthcare. The VPH Network of Excellence (VPH-NoE) project is tasked with facilitating interaction between the various VPH projects and addressing issues of common concern. A key deliverable is the 'VPH ToolKit'--a collection of tools, methodologies and services to support and enable VPH research, integrating and extending existing work across Europe towards greater interoperability and sustainability. Owing to the diverse nature of the field, a single monolithic 'toolkit' is incapable of addressing the needs of the VPH. Rather, the VPH ToolKit should be considered more as a 'toolbox' of relevant technologies, interacting around a common set of standards. The latter apply to the information used by tools, including any data and the VPH models themselves, and also to the naming and categorizing of entities and concepts involved. Furthermore, the technologies and methodologies available need to be widely disseminated, and relevant tools and services easily found by researchers. The VPH-NoE has thus created an online resource for the VPH community to meet this need. It consists of a database of tools, methods and services for VPH research, with a Web front-end. This has facilities for searching the database, for adding or updating entries, and for providing user feedback on entries. Anyone is welcome to contribute.

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.

Genital human papillomaviruses among women of reproductive age in Jamaica Virus de los papilomas humanos genitales en mujeres en edad reproductiva de Jamaica

Directory of Open Access Journals (Sweden)

Karen Lewis-Bell

2013-03-01

sexualmente activas, de 16 a 49 años de edad, que acudieron a uno de los consultorios públicos o privados de atención primaria seleccionados en cada una de las cuatro autoridades sanitarias regionales de Jamaica. Personal capacitado del estudio recopiló datos sociodemográficos de cada participante. Todas las participantes fueron sometidas a un examen ginecológico que comprendía una prueba clínica de Papanicolaou y la obtención de una muestra del cuello uterino a efectos de detectar y tipificarlos VPH mediante la prueba de genotipado Linear Array (LA (Roche Diagnostics Corp., Indianápolis, Indiana, Estados Unidos, de uso exclusivo en investigación. Se calcularon las prevalencias global y específica de tipo de la infección por VPH para los 37 tipos de VPH incluidos en la prueba de genotipado LA. RESULTADOS: Se detectó ADN de VPH en 460 de las 852 mujeres (54,0%. Se detectaron VPH oncógenos en 297 mujeres (34,9%, y VPH de los tipos 16 y 18 en 86 mujeres (10,1%. Los tipos de VPH detectados con mayor frecuencia fueron 16 (6,2%, 35 (6,0%, 62 y 83 (5,5%, 61 y 58 (5,4%, 84 (4,7%, 18 (4,3%, y 66 y 81 (4,2%. La prevalencia de VPH fue más elevada en mujeres solteras, jóvenes (de 16 a 19 años y que habían tenido más de tres compañeros sexuales en sus vidas. CONCLUSIONES: Estos resultados, junto a las elevadas tasas de cáncer cervicouterino, fundamentan la introducción de las vacunas contra el VPH al tiempo que se mantienen y refuerzan los servicios de tamizaje del cáncer cervicouterino. Las decisiones políticas que se adopten como consecuencia de estos resultados serán determinantes para establecer un programa integral contra el cáncer cervicouterino en Jamaica.

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...

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.

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.

Relación existente entre la infección por los diferentes genotipos del Virus del Papiloma Humano y la presencia de patología premaligna y maligna del cuello uterino

OpenAIRE

Mazarico Gallego, Edurne

2012-01-01

1) Introducción Hasta el momento se han secuenciado total o parcialmente más de 100 tipos y subtipos de VPH. De todos ellos, aproximadamente 40 tipos se han aislado en lesiones de tracto genital inferior y unos 35, según diferentes estudios, en carcinomas. Según su riesgo oncogénico, se clasifican en tipos de VPH de bajo riesgo (VPH-BR), VPH de probable alto riesgo y VPH de alto riesgo (VPH-AR). Uno de los descubrimientos más importantes en la investigación etiológica del cáncer en los...

Optimization of Highway Work Zone Decisions Considering Short-Term and Long-Term Impacts

Science.gov (United States)

2010-01-01

year when rehabilitation is conducted c0 vph Maximum discharge rate without work zone cw vph Maximum discharge rate with work zone CT $/project...vehicle class Q(t) vph Time-varying traffic flow volume Qp,max vph Maximum allowed diverted volume q(t) veh Time-varying number of vehicles in queue...and road capacity=1600 vph . Unlike single-regime Greenshield’s model, the traffic flow model converted from CORSIM car-following logic is multi

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

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...

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

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,...

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.

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.

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.

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...

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.

HPV vaginal self-sampling among women non-adherent to Papanicolaou screening in Chile Autotoma vaginal para detección de virus del papiloma humano en mujeres no adherentes a tamizaje con Papanicolaou en Chile

Directory of Open Access Journals (Sweden)

Javiera Léniz

2013-04-01

Full Text Available OBJECTIVE: To evaluate acceptance, preference and compliance with referral of vaginal self-sampling for the detection of Human papillomavirus (HPV among women non-adherent to Papanicolaou (Pap screening in Santiago, Chile. MATERIALS AND METHODS: Using multistage sampling we identified women aged 30-64 years who reported not receiving a Pap test in the previous three years and offered them Pap testing at the health center or vaginal self-sampling for HPV testing at home. Self-collected samples were analyzed with hybrid capture. All HPV+ women were referred for colposcopy, biopsy and treatment when needed. RESULTS: 1 254 eligible women were contacted; 86.5% performed self-sampling and 8.1% refused; 124 women were HPV+ (11.4%: 95%CI 9.6-13.5 of whom 85.5% attended colposcopy; 12 had CIN2+ (1.1%: 95 %CI 0.5-1.7. CONCLUSION: HPV vaginal self-sampling can be easily implemented in Chile and could improve coverage, successfully reaching women who drop out of the screening program.OBJETIVO: Evaluar la aceptación, preferencia y adherencia a seguimiento de la autotoma vaginal para detección del virus del papiloma humano (VPH en mujeres inasistentes a Papanicolaou (Pap en Santiago, Chile. MATERIAL Y MÉTODOS: Mediante un muestreo polietápico se identificaron mujeres entre 30 y 64 años inasistentes a Pap por < 3 años, invitándolas a realizarse un Pap en su centro de salud o una autotoma vaginal a domicilio. Las muestras fueron analizadas con captura de híbridos. Las mujeres VPH+ fueron referidas a colposcopía, biopsia y tratamiento en caso necesario. RESULTADOS: 1 254 mujeres elegibles fueron contactadas; 86.5% aceptó la autotoma vaginal y 8.1% la rechazó; 124 mujeres resultaron VPH+ (11.4%: IC95% 9.6-13.5 de las que 85.5% asistió a colposcopía; 12 tenían CIN2+ (1.1%: IC95% 0.5-1.7. CONCLUSIÓN: La autotoma vaginal para detección de VPH es implementable en Chile y su utilización podría mejorar la cobertura del programa rescatando a mujeres

Use of Serum Bicarbonate to Substitute for Venous pH in New-Onset Diabetes.

Science.gov (United States)

von Oettingen, Julia; Wolfsdorf, Joseph; Feldman, Henry A; Rhodes, Erinn T

2015-08-01

To investigate whether serum bicarbonate (HCO3) levels can be used to accurately diagnose diabetic ketoacidosis (DKA) and classify its severity in children with new-onset diabetes mellitus (NODM). Retrospective study of all patients with NODM presenting to Boston Children's Hospital from October 1, 2007, to July 1, 2013. DKA was defined as blood glucose ≥200 mg/dL, venous pH (vpH) vpH vpH, and logistic regression to evaluate serum HCO3 as a predictor of DKA and severe DKA. Of 690 study cohort subjects (47% girls, age 10.8 ± 4.3 years, 76.7% white), 19.4% presented with DKA. The relationship between serum HCO3 and vpH was log-linear (r = 0.87, 95% CI 0.85-0.89, P vpH (R(2) 0.75, P vpH = 6.81301 + (0.17823*ln[HCO3]) and DKA and severe DKA (c-statistic 0.97 [95% CI 0.96-0.99, P vpH to diagnose DKA and classify severity in children with NODM. It is suggested as an alternative to reliance on vpH, especially in settings in which access to vpH measurement is limited. Copyright © 2015 by the American Academy of Pediatrics.

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.

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.

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...

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

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.

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

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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.

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

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

Azimuthal Spoke Propagation in Hall Effect Thrusters

Science.gov (United States)

2013-10-01

group velocity, m s−1 vph = phase velocity, m s−1 vs = ion acoustic velocity, m s−1 vsp = spoke velocity, m s−1 vspj,k = spoke velocity from bin n to m...phase velocity, vph , and group velocity, vgr, from the dispersion relation in Eq. (7) are vph = ω kθ = [ vαch − ( ωch kθ )α]1/α (9) vgr = ∂ω ∂kθ = vph ...vch vph )α (10) Eq. (9) shows that the phase velocity will always be less than the characteristic velocity and Eq. (10) shows the group velocity will

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

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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.

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...

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.)

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

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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.

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...

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.

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 ...

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

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

Veterinary public health in India: current status and future needs.

Science.gov (United States)

Ghatak, S; Singh, B B

2015-12-01

Veterinary public health (VPH) assumes huge significance in developing countries such as India. However, the implementation of VPH services throughout the country is still in its infancy. From 1970 onwards, many institutes, national and international organisations, professional societies, policies and personalities have contributed towards the development of VPH in India. Nevertheless, there is an urgent need to develop VPH still further as there are many issues, such as high population density, the re-emergence of zoonotic pathogens, environmental pollution and antimicrobial resistance, that require attention. The time has surely come to involve all stakeholders, ranging from primary producers (e.g., farmers) to policy-makers, so as to garner support for the holistic implementation of VPH services in India. To improve VPH activities and services, science-based policies enforced through stringent regulation are required to improve human, animal and environmental health. The emergence of the 'One Health' concept has ushered in new hopes for the resurrection of VPH in India. Applying tools such as the World Organisation for Animal Health (OlE) Day One Competencies and the OlE Tool for the Evaluation of Performance of Veterinary Services (PVS Tool) is essential to improve the quality of national Veterinary Services and to identify gaps and weaknesses in service provision, which can be remedied to comply with the OlE international standards. VPH initiatives started modestly but they continue to grow. The present review is focused on the current status and future needs of VPH in India.

Determination of the Ionosphere Parameters by Analyzing the Propagation After-Effects

Science.gov (United States)

2010-06-27

whereas the long waves are subject to stronger dispersion. Indeed, the phase and group velocity of the propagation are given by: vph = c ( 1 + ω2pe/c...antenna at x is given by equation (A.16) [cf. formula (2.15)]: ϕ(t, z ) = A′(t− |z − x |/vgr(ω0)) 4π|z − x | eiω0(t−|z−x |/ vph (ω0)), where A′(t) = χτ ′(t...carrier oscillation per se, i.e., the wave eiω0t, travels with the corresponding phase velocity vph (ω0) that can also be linearized: vph = vph (ω) ≈ c ( 1

Open Architecture Framework for Improved Early Stage Submarine Design

Science.gov (United States)

2010-06-01

Figure 12 Data Structure Example – VPH ..................................................................................... 41 Figure 13 SUBSTART Data...in the MIT SMM (e.g. – VPH -VB, pressure hull volume without volume of the in-hull variable ballast). Tables 4 and 5 list the variables by module...MUD1frac FFsurf θ(x,t) V(t) VPH ∆ff VCGLEADs BTf MUDfwd GMt R(x) Vambt VPHguess ∆mbt VCGnsc BTops OBambt ηa tenvsurf Vaux VPH -VB ∆nsc VCGVL

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.

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...

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

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

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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.

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

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

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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

Verrugas plantares: caracterización de los virus causales y aplicación del láser 1064 nm a su tratamiento

OpenAIRE

Planell Mas, Elena de

2016-01-01

INTRODUCCIÓN: Las verrugas plantares están provocadas por el virus del papilloma humano (VPH). Se han descrito diferentes genotipos asociados a estas lesiones si bien existen pocos estudios centrados exclusivamente en los genotipos de las verrugas plantares. El diagnóstico generalmente es clínico, aunque actualmente existen métodos diagnósticos basados en la detección del ADN viral como es el caso de la PCR (polymerase chain reaction). Las modalidades de tratamiento de las verrugas so...

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

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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.

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...

Inverse fusion PCR cloning.

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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.

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.

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.

Quantitative Analysis for Installation Access Planning at Naval Base San Diego

Science.gov (United States)

2012-09-01

VPH in one processing lane, then we can assume that if the SECO were to open another sentry processing lane, the total throughput of both lanes would...be 600 VPH . Similarly, if two sentries in tandem can produce a throughput of 500 VPH , then having two lanes with two sentries in tandem each will...produce a total throughput of 1000 VPH . We assume throughout that there is no server idleness and so there is essentially an infinite backlog of

Effect of biochar or activated carbon amendment on the volatilisation and biodegradation of organic soil pollutants

Science.gov (United States)

Werner, David; Meynet, Paola; Bushnaf, Khaled

2013-04-01

Biochar or activated carbon added to contaminated soil may temporarily reduce the volatilisation of organic pollutants by enhanced sorption. The long-term effect of sorbent amendments on the fate of volatile petroleum hydrocarbon mixtures (VPHs) will depend on the responses of the soil bacterial community members, especially those which may utilize VPHs as carbon substrates. We investigated the volatilisation and biodegradation of VPHs emanating from NAPL sources and migrating through one meter long columns containing unsaturated sandy soil with and without 2% biochar or activated carbon amendment. After 420 days, VPH volatilisation from AC amended soil was less than 10 percent of the cumulative VPH volatilisation flux from unamended soil. The cumulative CO2 volatilisation flux increased more slowly in AC amended soil, but was comparable to the untreated soil after 420 days. This indicated that the pollution attenuation over a 1 meter distance was improved by the AC amendment. Biochar was a weaker VPH sorbent than AC and had a lesser effect on the cumulative VPH and CO2 fluxes. We also investgated the predominant bacterial community responses in sandy soil to biochar and/or VPH addition with a factorially designed batch study, and by analyzing preserved soil samples. Biochar addition alone had only weak effects on soil bacterial communities, while VPH addition was a strong community structure shaping factor. The bacterial community effects of biochar-enhanced VPH sorption were moderated by the limited biomass carrying capacity of the sandy soil investigated which contained only low amounts of inorganic nitrogen. Several Pseudomonas spp., including Pseudomonas putida strains, became dominant in VPH polluted soil with and without biochar. The ability of these versatile VPH degraders to effectively regulate their metabolic pathways according to substrate availabilities may additionally have moderated bacterial community structure responses to the presence of biochar

A vision and strategy for the virtual physiological human: 2012 update.

Science.gov (United States)

Hunter, Peter; Chapman, Tara; Coveney, Peter V; de Bono, Bernard; Diaz, Vanessa; Fenner, John; Frangi, Alejandro F; Harris, Peter; Hose, Rod; Kohl, Peter; Lawford, Pat; McCormack, Keith; Mendes, Miriam; Omholt, Stig; Quarteroni, Alfio; Shublaq, Nour; Skår, John; Stroetmann, Karl; Tegner, Jesper; Thomas, S Randall; Tollis, Ioannis; Tsamardinos, Ioannis; van Beek, Johannes H G M; Viceconti, Marco

2013-04-06

European funding under Framework 7 (FP7) for the virtual physiological human (VPH) project has been in place now for 5 years. The VPH Network of Excellence (NoE) has been set up to help develop common standards, open source software, freely accessible data and model repositories, and various training and dissemination activities for the project. It is also working to coordinate the many clinically targeted projects that have been funded under the FP7 calls. An initial vision for the VPH was defined by the FP6 STEP project in 2006. In 2010, we wrote an assessment of the accomplishments of the first two years of the VPH in which we considered the biomedical science, healthcare and information and communications technology challenges facing the project (Hunter et al. 2010 Phil. Trans. R. Soc. A 368, 2595-2614 (doi:10.1098/rsta.2010.0048)). We proposed that a not-for-profit professional umbrella organization, the VPH Institute, should be established as a means of sustaining the VPH vision beyond the time-frame of the NoE. Here, we update and extend this assessment and in particular address the following issues raised in response to Hunter et al.: (i) a vision for the VPH updated in the light of progress made so far, (ii) biomedical science and healthcare challenges that the VPH initiative can address while also providing innovation opportunities for the European industry, and (iii) external changes needed in regulatory policy and business models to realize the full potential that the VPH has to offer to industry, clinics and society generally.

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

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.

Gluteoplastia tridimensional mediante distribución volumétrica precisa

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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.

A vision and strategy for the virtual physiological human in 2010 and beyond.

Science.gov (United States)

Hunter, Peter; Coveney, Peter V; de Bono, Bernard; Diaz, Vanessa; Fenner, John; Frangi, Alejandro F; Harris, Peter; Hose, Rod; Kohl, Peter; Lawford, Pat; McCormack, Keith; Mendes, Miriam; Omholt, Stig; Quarteroni, Alfio; Skår, John; Tegner, Jesper; Randall Thomas, S; Tollis, Ioannis; Tsamardinos, Ioannis; van Beek, Johannes H G M; Viceconti, Marco

2010-06-13

European funding under framework 7 (FP7) for the virtual physiological human (VPH) project has been in place now for nearly 2 years. The VPH network of excellence (NoE) is helping in the development of common standards, open-source software, freely accessible data and model repositories, and various training and dissemination activities for the project. It is also helping to coordinate the many clinically targeted projects that have been funded under the FP7 calls. An initial vision for the VPH was defined by framework 6 strategy for a European physiome (STEP) project in 2006. It is now time to assess the accomplishments of the last 2 years and update the STEP vision for the VPH. We consider the biomedical science, healthcare and information and communications technology challenges facing the project and we propose the VPH Institute as a means of sustaining the vision of VPH beyond the time frame of the NoE.

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)

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)

Human papillomavirus genotypes in invasive cervical squamous cell carcinoma in Trinidad Genotipos de virus de los papilomas humanos en carcinoma cervicouterino escamocelular invasor en Trinidad

Directory of Open Access Journals (Sweden)

Felicia Hosein

2013-04-01

Full Text Available OBJECTIVE: To determine the relative contribution of known high-risk human papillomavirus (HPV genotypes to the occurrence of cervical cancers in Trinidad. METHODS: The distribution of HPV genotypes in cases of invasive cervical squamous cell carcinoma in Trinidad was investigated. This study was a follow-up to an investigation of HPV genotypes in 310 nonsymptomatic women in Trinidad. The latter study showed that cervical HPV prevalence and heterogeneity of genotypes were high in the study population; notably, the genotypes targeted by the available HPV prophylactic vaccines were not the most common types. RESULTS: The current study of 85 cases of invasive cervical squamous cell carcinomas demonstrated that the previously observed heterogeneity in HPV genotype distribution is lost in cases of invasive cervical cancer, with the vaccine-targeted HPV types HPV 16 and HPV 18 becoming the most prevalent. CONCLUSIONS: HPV 16 and HPV 18 were the primary HPV genotypes associated with cases of invasive squamous cell carcinoma in the current Trinidad study. This strong association leads us to conclude that the HPV vaccines targeting HPV 16 and HPV 18 may contribute to reducing the cervical cancer burden in Trinidad.OBJETIVO: Determinar la contribución relativa de los diferentes genotipos de virus de los papilomas humanos (VPH conocidos como de alto riesgo para la aparición de cáncer cervicouterino en Trinidad. MÉTODOS: Se investigó la distribución de los genotipos de VPH en casos de carcinoma cervicouterino escamocelular invasor en Trinidad. Este estudio fue la continuación de una investigación de los genotipos de VPH presentes en 310 mujeres asintomáticas en Trinidad. Este último estudio reveló altas prevalencia de VPH en el cuello uterino y heterogeneidad de los genotipos en la población del estudio; cabe destacar que los genotipos a los que se dirigen las vacunas preventivas de la infección por VPH disponibles no fueron los tipos m

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

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...

Thermally Stable Siloxane Hybrid Matrix with Low Dielectric Loss for Copper-Clad Laminates for High-Frequency Applications.

Science.gov (United States)

Kim, Yong Ho; Lim, Young-Woo; Kim, Yun Hyeok; Bae, Byeong-Soo

2016-04-06

We report vinyl-phenyl siloxane hybrid material (VPH) that can be used as a matrix for copper-clad laminates (CCLs) for high-frequency applications. The CCLs, with a VPH matrix fabricated via radical polymerization of resin blend consisting of sol-gel-derived linear vinyl oligosiloxane and bulky siloxane monomer, phenyltris(trimethylsiloxy)silane, achieve low dielectric constant (Dk) and dissipation factor (Df). The CCLs with the VPH matrix exhibit excellent dielectric performance (Dk = 2.75, Df = 0.0015 at 1 GHz) with stability in wide frequency range (1 MHz to 10 GHz) and at high temperature (up to 275 °C). Also, the VPH shows good flame resistance without any additives. These results suggest the potential of the VPH for use in high-speed IC boards.

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

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.

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.

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

Doping Optimization for High Efficiency in Semiconductor Diode Lasers and Amplifiers

Science.gov (United States)

2016-03-01

ηiVph αm αm + αi (I − Ith) , (12) where ηi is the internal quantum efficiency, Vph is the voltage associated with the energy of a single photon, and...efficiency. In general, this could be a negligible detail; however, for certain cases such as V0 Vph , the difference could be significant, since...communications applications. Bour and Rosen provided an expression for the maximum PCE of a diode laser, given as [12] ηPCE = ηi Vph V0 αm αm + αi x( 1 + √ 1 + x

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...

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

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

Caracterización por PCR- múltiple del grupo filogenético de Escherichia coli uropatógena aisladas de pacientes ambulatorios de Bucaramanga, Santander

Directory of Open Access Journals (Sweden)

Alexandra Serrano

2016-06-01

Full Text Available Introducción: Las infecciones del tracto urinario son consideradas actualmente como una de las enfermedades infecciosas más comunes. En general se acepta que los agentes infecciosos con los cuales se asocia se encuentran presentes en la microbiota intestinal, por tal razón se ha descrito que Escherichia coli es una de las principales Enterobacterias involucradas en las manifestaciones clínicas presentes en pacientes con infecciones del tracto urinario. Las cepas de E. coli han sido clasificadas inicialmente en 4 grupos filogenéticos según Clermont y col. (2000, de este modo se conocen “cepas intestinales comensales, pertenecientes a los grupos A y B1, y cepas extra intestinales virulentas asociadas a los grupos B2 y D, para llevar a cabo dicha clasificación Clermont, diseñó herramientas moleculares como la PCR-triple, mediante la utilización de diferentes genes tales como ChuA, yjaA y TSPE4.C2”. Sin embargo debido a la complejidad estructural genética de este microorganismo, se han logrado establecer nuevos grupos filogenéticos de E. coli, A, B1, B2, C, D, E, F y un último grupo el cual no ha sido completamente identificado, por tal razón es importante conocer cuáles son los grupos filogenéticos que están presentes en nuestra región, mediante el uso de técnicas moleculares que permitan realizar la caracterización filogenética de dicho microorganismo y de esta manera lograr establecer los perfiles de resistencia y evaluar los posibles factores de virulencia asociados al proceso de infección. Objetivo: Realizar la caracterización molecular de los grupos filogenéticos de E. coli uropatógena aislada de pacientes ambulatorios que asisten a un laboratorio de tercer nivel de complejidad de la ciudad de Bucaramanga mediante el uso de PCR-multiplex. Materiales y métodos: El cálculo del tamaño muestral se realizó teniendo en cuenta la prevalencia reportada por Orduz, K y Trejos, J, (2010, se calculó el tamaño de muestra

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.

Virtual physiological human: training challenges.

Science.gov (United States)

Lawford, Patricia V; Narracott, Andrew V; McCormack, Keith; Bisbal, Jesus; Martin, Carlos; Bijnens, Bart; Brook, Bindi; Zachariou, Margarita; Freixa, Jordi Villà I; Kohl, Peter; Fletcher, Katherine; Diaz-Zuccarini, Vanessa

2010-06-28

The virtual physiological human (VPH) initiative encompasses a wide range of activities, including structural and functional imaging, data mining, knowledge discovery tool and database development, biomedical modelling, simulation and visualization. The VPH community is developing from a multitude of relatively focused, but disparate, research endeavours into an integrated effort to bring together, develop and translate emerging technologies for application, from academia to industry and medicine. This process initially builds on the evolution of multi-disciplinary interactions and abilities, but addressing the challenges associated with the implementation of the VPH will require, in the very near future, a translation of quantitative changes into a new quality of highly trained multi-disciplinary personnel. Current strategies for undergraduate and on-the-job training may soon prove insufficient for this. The European Commission seventh framework VPH network of excellence is exploring this emerging need, and is developing a framework of novel training initiatives to address the predicted shortfall in suitably skilled VPH-aware professionals. This paper reports first steps in the implementation of a coherent VPH training portfolio.

Experimental and Analytical Research on Resonance Phenomena of Vibrating Head with MRE Regulating Element

Science.gov (United States)

Miedzińska, D.; Gieleta, R.; Osiński, J.

2015-02-01

A vibratory pile hammer (VPH) is a mechanical device used to drive steel piles as well as tube piles into soil to provide foundation support for buildings or other structures. In order to increase the stability and the efficiency of the VPH work in the over-resonance frequency, a new VPH construction was developed at the Military University of Technology. The new VPH contains a system of counter-rotating eccentric weights, powered by hydraulic motors, and designed in such a way that horizontal vibrations cancel out, while vertical vibrations are transmitted into the pile. This system is suspended in the static parts by the adaptive variable stiffness pillows based on a smart material, magnetorheological elastomer (MRE), whose rheological and mechanical properties can be reversibly and rapidly controlled by an external magnetic field. The work presented in the paper is a part of the modified VPH construction design process. It concerns the experimental research on the vibrations during the piling process and the analytical analyses of the gained signal. The results will be applied in the VPH control system.

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...

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.

PCR em tempo real para diagnóstico da leucose enzoótica bovina Enzootic bovine leukosis real time PCR

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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

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)

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.

Realidad y nuevos horizontes de la vacuna del virus del papiloma humano

OpenAIRE

García Gómez, Nuria

2014-01-01

La vacuna frente al VPH es la primera vacuna destinada a la prevención del cáncer inducido por un virus, el CCU (problema de salud pública mundial), y de lesiones precursoras. Disponemos de 2 vacunas profilácticas: Gardasil® (frente a VPH 6, 11, 16 y 18) y Cervarix® (frente a VPH 16 y 18). El trabajo consiste en una revisión bibliográfica que tiene como objetivo conocer la situación que vive actualmente la vacuna del VPH en España, utilizando como palabras clave virus, vacun...

El virus del Papiloma Humano en cavidad oral y orofaríngea en estudiantes universitarios de 18 a 25 años en Valencia

OpenAIRE

Sastre Cantón, Macrina

2017-01-01

INTRODUCCIÓN Y JUSTIFICACIÓN DEL ESTUDIO El VPH es una de las infecciones virales más extendidas en la población. Estudios epidemiológicos revelan que la infección oral por VPH juega un papel importante en el desarrollo de enfermedades precancerosas o cánceres de la cavidad oral, en particular en el cáncer de células escamosas orales.1 Aunque se han descrito más de 140 genotipos de VPH, no todos ellos tienen el mismo riesgo de provocar lesiones pre-malignas o cánceres. El genotipo VPH-16 ...

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.

La infección por el virus del papiloma humano, un posible marcador biológico de comportamiento sexual en estudiantes universitarios

Directory of Open Access Journals (Sweden)

Sánchez-Alemán Miguel A

2002-01-01

Full Text Available Objetivo. Estimar la prevalencia de infección por el virus del papiloma humano (VPH en estudiantes universitarios y utilizar dicha frecuencia como un marcador biológico para evaluar el comportamiento sexual. Material y métodos. Se realizó un estudio transversal, en estudiantes de la Universidad Autónoma del estado de Morelos, México, durante el periodo 2000-2001. Se aplicó un cuestionario y se colectaron muestras genitales para detectar ADN de los VPH oncogénicos. Los datos se analizaron utilizando pruebas de Ji cuadrada y razones de momios. Resultados. La prevalencia global del VPH en 194 estudiantes fue de 14.4%. Las mujeres con dos o más parejas sexuales durante el último año presentaron mayor riesgo de infección por el VPH (RM 6.0 IC 1.7-21.1, al igual que las que utilizaron anticonceptivos hormonales y espermicidas en su última relación sexual (RM 3.0 IC 1.0-8.7. Los hombres que consumieron cocaína tuvieron más riesgo de infección por el VPH (RM 7.6 IC 1.3-45.1. Conclusiones. La prevalencia del VPH es relativamente alta. La utilización del VPH como un marcador biológico de comportamientos sexuales en mujeres es pertinente; en hombres, es necesario ampliar la muestra.

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.

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.

Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR for detection of avian metapneumovirus subtype A Comparação entre as técnicas de RT-PCR convencional e RT-PCR em tempo real para a detecção do metapneumovírus aviários subtipo A

Directory of Open Access Journals (Sweden)

Helena Lage Ferreira

2009-08-01

Full Text Available 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 established test for the attachment (G gene. All the RT-PCR tested assays were able to detect the AMPV/A. The lower detection limits were observed using the N-, F- based RRT-PCR and F-based conventional RT-PCR (10(0.3 to 10¹ TCID50 mL-1. The present study suggests that the conventional F-based RT-PCR presented similar detection limit when compared to N- and F-based RRT-PCR and they can be successfully used for AMPV/A detection.O metapneumovírus aviário (AMPV pertence ao gênero Metapneumovirus, família Paramyxoviridae. Isolamento viral, sorologia e detecção do RNA genômico são atualmente as técnicas utilizadas para o diagnóstico desse agente. O objetivo do presente estudo foi comparar a detecção de RNA viral de seis isolados de AMPV, subtipo A (AMPV/A, utilizando diferentes métodos de RT-PCR convencional e real time RT-PCR (RRT-PCR. Duas novas técnicas de RT-PCR convencional e duas técnicas de RRT-PCR, ambas para a detecção dos genes da nucleoproteína (N e da proteína de fusão (F, foram comparadas com um RT-PCR previamente estabelecido para a detecção do AMPV (gene da glicoproteína -G. Todos esses métodos foram capazes de detectar os isolados AMPV/A. As técnicas RRT-PCR (genes F e N mostraram os menores limites de detecção (10(0.3 to 10¹ TCID50 mL-1. Os resultados sugerem que as técnicas RT-PCR convencional (gene F e as técnicas de RRT-PCR (gene F e N desenvolvidas no presente estudo podem ser utilizadas com sucesso para a detecção do

Mise à jour sur le nouveau vaccin 9-valent pour la prévention du virus du papillome humain

Science.gov (United States)

Yang, David Yi; Bracken, Keyna

2016-01-01

Résumé Objectif Informer les médecins de famille quant à l’efficacité, à l’innocuité, aux effets sur la santé publique et à la rentabilité du vaccin 9-valent contre le virus du papillome humain (VPH). Qualité des données Des articles pertinents publiés dans PubMed jusqu’en mai 2015 ont été examinés et analysés. La plupart des données citées sont de niveau I (essais randomisés et contrôlés et méta-analyses) ou de niveau II (études transversales, cas-témoins et épidémiologiques). Des rapports et recommandations du gouvernement sont aussi cités en référence. Message principal Le vaccin 9-valent contre le VPH, qui offre une protection contre les types 6, 11, 16, 18, 31, 33, 45, 52 et 58 du VPH, est sûr et efficace et réduira encore plus l’incidence des infections à VPH, de même que les cas de cancer lié au VPH. Il peut également protéger indirectement les personnes non immunisées par l’entremise du phénomène d’immunité collective. Un programme d’immunisation efficace peut prévenir la plupart des cancers du col de l’utérus. Les analyses montrent que la rentabilité du vaccin 9-valent chez les femmes est comparable à celle du vaccin quadrivalent original contre le VPH (qui protège contre les types 6, 11, 16 et 18 du VPH) en usage à l’heure actuelle. Toutefois, il faut investiguer plus en profondeur l’utilité d’immuniser les garçons avec le vaccin 9-valent contre le VPH. Conclusion en plus d’être sûr, le vaccin 9-valent protège mieux contre le VPH que le vaccin quadrivalent. Une analyse coûtefficacité en favorise l’emploi, du moins chez les adolescentes. Ainsi, les médecins devraient recommander le vaccin 9-valent à leurs patients plutôt que le vaccin quadrivalent contre le VPH.

The Spanish human papillomavirus vaccine consensus group: a working model.

Science.gov (United States)

Cortés-Bordoy, Javier; Martinón-Torres, Federico

2010-08-01

Successful implementation of Human Papillomavirus (HPV) vaccine in each country can only be achieved from a complementary and synergistic perspective, integrating all the different points of view of the diverse related professionals. It is this context where the Spanish HPV Vaccine Consensus Group (Grupo Español de Consenso sobre la Vacuna VPH, GEC-VPH) was created. GEC-VPH philosophy, objectives and experience are reported in this article, with particular attention to the management of negative publicity and anti-vaccine groups. Initiatives as GEC-VPH--adapted to each country's particular idiosyncrasies--might help to overcome the existing barriers and to achieve wide and early implementation of HPV vaccination.

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...

Ensayo no destructivo de soldaduras en pernos conectores mediante inspección acústica

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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

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...

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.

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.

PCR en tiempo real: una metodología útil para la detección y cuantificación de granulovirus

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Gloria Patricia Barrera Cubillos

2016-07-01

Full Text Available El uso de baculovirus como agentes de control biológico de insectos plaga, se ha convertido en una estrategia efectiva que se ha implementado gradualmente en diferentes sistemas productivos a nivel mundial. Para el desarrollo de un bioplaguicida a base de baculovirus, es necesario contar con una metodología para determinar el título viral en el producto en proceso y terminado. Para tal fin, en este trabajo se diseñó y optimizó una técnica de cuantificación viral (Betabaculovirus mediante PCR cuantitativo (q-PCR. Se utilizó una sonda TaqMan diseñada sobre el gen de granulina, altamente conservado. Para la técnica de q-PCR se determinó la especificidad, sensibilidad y reproducibilidad, encontrando que puede detectar y cuantificar aislamientos del género Betabaculovirus provenientes de cinco especies diferentes de insectos (granulovirus de Tecia solanivora, Phthorimaea operculella, Erinnyis ello, Tuta absoluta y Spodoptera frugiperda incluso de diferente origen geográfico, pero no detecta aislamientos del género Alphabaculovirus (nucleopoliedrovirus de Spodoptera ornithogalli, Diatraea saccharalis o S. frugiperda. El límite mínimo de detección de la técnica fue de 6,4 x 10-4 ng de ADN, lo que equivale a 1,25 x 105 copias del gen. Así mismo, la variación intra e inter ensayos fue mínima, demostrando la reproducibilidad de la misma. La aplicabilidad de la técnica fue evaluada para la detección de granulovirus en muestras de larva, suelo, y para determinar la concentración viral en un bioplaguicida formulado como concentrado emulsionable. En conclusión, la técnica de q-PCR desarrollada fue reproducible, sensible y específica, con aplicabilidad en estudios de persistencia viral en campo, control de infecciones en crías de insectos y control de calidad de bioplaguicidas a base de betabaculovirus.

Prospects for primary prevention of cervical cancer in developing countries Perspectivas de prevención primaria de cáncer cervical en países en desarrollo

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Silvia Franceschi

2003-01-01

mediante la conducción de ensayos clínicos aleatorizados. El primer tipo de estudios no puede demostrar en forma convincente su efectividad, porque las tasas de incidencia y mortalidad por cáncer cervical son influenciadas fuertemente por tendencias seculares que son independientes de las medidas de intervención. Estudios de fase 3 de vacunas profilácticas contra el VPH están siendo desarrollados actualmente. Cada uno de los ensayos es muy costoso e involucran complejos tipos de diseño. Esto es importante, sin embargo, deben iniciarse, tan pronto como sea posible, ensayos más simples diseñados para demostrar la efectividad de una vacuna contra el VPH en condiciones de campo, como las que pueden existir en países en desarrollo, donde el peso de la mortalidad por cáncer cervical es muy alto. Cada ensayo clínico puede cuantificar una diferencia en la presentación de lesiones precursoras de cáncer (el real blanco de una vacuna contra el VPH en un periodo de seguimiento prolongado (20 años, al menos. En este artículo se presenta brevemente el posible diseño de este tipo de estudios, considerando dos áreas: India meridional y Corea del Sur.

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

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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.

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.

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

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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

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...

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.

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...

Factores de riesgo de cáncer cervicouterino invasor en mujeres mexicanas Risk factors in invasive cervical cancer among Mexican women

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Laura Leticia Tirado-Gómez

2005-10-01

Full Text Available OBJETIVOS: Evaluar la asociación entre cáncer cervicouterino (CaCu invasor y el virus del papiloma humano de alto riesgo (VPH-AR (carga viral/tipo 16, así como con factores ginecológicos y socioeconómicos. MATERIAL Y MÉTODOS: Estudio de casos y controles pareado individualmente (215 casos con CaCu invasor y 420 controles. La población de estudio se reclutó entre los años 2000 y 2001. Se evaluaron variables tradicionalmente asociadas con CaCu (ginecológicas y socioeconómicas y dos variables asociadas con la presencia de VPH (carga viral y el tipo 16. La presencia de VPH-AR se determinó mediante Captura de Híbridos II. La carga viral se midió a través de unidades relativas de luz y picogramos por ml (1 RLU=1 pg/ml, divididas en cuatro categorías: negativa (499 pg/ml. El análisis estimó razones de momios (RM ajustadas a través de modelos de regresión logística condicionada. RESULTADOS: La presencia de VPH-AR incrementa en 78 veces la probabilidad de presentar CaCu invasor; cuando el VPH es tipo 16, el incremento es mayor (RM= 429.7 comparado con otros tipos (RM=64.1. Se observó una tendencia importante en la RM al elevarse la carga viral (RM=46.6 carga baja; RM=250.7 intermedia y RM=612.9 alta. Finalmente, los factores demográficos y obstétricos conocidos, incrementaron la probabilidad de CaCu invasor. No se observó asociación entre CaCu invasor y tabaquismo en la población de estudio. CONCLUSIONES: Este estudio contribuye a la identificación de las mujeres con alto riesgo de desarrollar CaCu invasor, entre las pacientes infectadas con VPH-AR. Por otra parte, confirma la importancia de la infección de VPH-AR y refleja la carga viral del VPH-AR como cofactor y posible promotor en el desarrollo de la enfermedad. Por último, este biomarcador puede contribuir a mejorar la prevención y la detección temprana de esta enfermedad.OBJECTIVE: To evaluate the association between invasive Cervical Cancer (CC and high risk Human

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.

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

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

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

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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

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

Transgene detection by digital droplet PCR.

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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.

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.

[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

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.

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.

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...

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...

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.

Visceral pain hypersensitivity in functional gastrointestinal disorders.

Science.gov (United States)

Farmer, A D; Aziz, Q

2009-01-01

Functional gastrointestinal disorders (FGIDs) are a highly prevalent group of heterogeneous disorders whose diagnostic criteria are symptom based in the absence of a demonstrable structural or biochemical abnormality. Chronic abdominal pain or discomfort is a defining characteristic of these disorders and a proportion of patients may display heightened pain sensitivity to experimental visceral stimulation, termed visceral pain hypersensitivity (VPH). We examined the most recent literature in order to concisely review the evidence for some of the most important recent advances in the putative mechanisms concerned in the pathophysiology of VPH. VPH may occur due to anomalies at any level of the visceral nociceptive neuraxis. Important peripheral and central mechanisms of sensitization that have been postulated include a wide range of ion channels, neurotransmitter receptors and trophic factors. Data from functional brain imaging studies have also provided evidence for aberrant central pain processing in cortical and subcortical regions. In addition, descending modulation of visceral nociceptive pathways by the autonomic nervous system, hypothalamo-pituitary-adrenal axis and psychological factors have all been implicated in the generation of VPH. Particular areas of controversy have included the development of efficacious treatment of VPH. Therapies have been slow to emerge, mainly due to concerns regarding safety. The burgeoning field of genome wide association studies may provide further evidence for the pleiotropic genetic basis of VPH development. Tangible progress will only be made in the treatment of VPH when we begin to individually characterize patients with FGIDs based on their clinical phenotype, genetics and visceral nociceptive physiology.

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 ...

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.

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

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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.

Development of Nested PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Cylindrocladium scoparium on Eucalyptus

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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.

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

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.

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...

PCR-RFLP y RAPD para la tipificación de Leishmania neotropical

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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.

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

RT-PCR Protocols - Methods in Molecular Biology

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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...........

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...

Comportamiento del tamizaje para cancer de cervix en la ciudad de Santa Marta en el año 2005

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José Abraham Jaramillo Osorio

2013-10-01

Full Text Available ResumenObjetivo: analizar los resultados del programa de detección y control de cáncer de cuello uterino en la ciudad de santa marta, Colombia durante el año 2005. Materiales y métodos: la información se obtuvo compilando los registros mensuales del laboratorio de citología de la E.S.E Alejandro Prospero Reverend en Santa Sarta durante el año 2005. Los datos fueron registrados en un archivo excel y luego evaluados mediante el programa Epi-info 2002. Resultados: durante el 2005 se realizaron en los distintos centros y puestos de salud 16683 citologías, 552 fueron inadecuadas (3,3% 534 alteradas (3,2% 340 infección por virus del papiloma humano (V.P.H. (2% 111 casos de lesión intraepitelial de bajo grado (lieb(0,67% 63 casos de lesión intraepitelial de alto grado(0,38% 15 casos de cáncer invasor de cuello uterino(89.9x100.000. En la vereda de machete y el corregimiento de Guachaca el promedio de infección por V.P.H. fue el doble comparado con los restantes. Conclusiones: este análisis permite identificar debilidades en cuanto a la cobertura y el suministro de información del programa de detección y control de cáncer de cuello uterino, además aporta datos de gran utilidad para diseñar estrategias en salud pública tendientes a corregir indicadores epidemiológicos. (Duazary 2006; 1: 32- 37

Use of On-Site GC/MS Analysis to Distinguish Between Vapor Intrusion and Indoor Sources of VOCs

Science.gov (United States)

2013-11-01

Petroleum Hydrocarbons (ETPH), MA-EPH, MA- VPH . Microbiology Parameters: Total Coliform – MF mEndo (SM9222B), Total Coliform – MTF (SM9221B), E. Coli...Parameters: PCBs, PCBs in Oil, Organochlorine Pesticides, Technical Chlordane, Toxaphene, CT-Extractable Petroleum Hydrocarbons (ETPH), MA-EPH, MA- VPH ...Organic Parameters: 608, 624, 625, 8081A, 8082, 8330, 8151A, 8260B, 8270C, 3510C, 3630C, 5030B, ME- DRO, ME-GRO, MA-EPH, MA- VPH .) Solid Waste/Soil

Environmental Assessment for Mid-Bay Bridge Connector, Eglin Air Force Base, Florida

Science.gov (United States)

2008-12-05

Year Average Speed (mph) AADT VPH Receptor Max 1-Hr Conc (ppm) Max 8-Hr Conc (ppm) Proposed Action 2001 45 17,400 981 Proposed...Speed (mph) AADT VPH Receptor Max 1-Hr Conc (ppm) Max 8-Hr Conc (ppm) Alternative C 2001 45 17,400 981 Proposed Intersection at SR 20...Alternative Traffic Volumes Alternative Year Average Speed White Point Road (mph) AADT VPH Receptor Max 1-Hr Conc (ppm) Max 8-Hr Conc (ppm

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...

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

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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.

[Molecular genetics in chronic myeloid leukemia with variant Ph translocation].

Science.gov (United States)

Wu, Wei; Li, Jian-yong; Zhu, Yu; Qiu, Hai-rong; Pan, Jin-lan; Xu, Wei; Chen, Li-juan; Shen, Yun-feng; Xue, Yong-quan

2007-08-01

To explore the value of fluorescence in situ hybridization (FISH) and multiplex fluorescence in situ hybridization (M-FISH) techniques in the detection of genetic changes in chronic myeloid leukemia (CML) with variant Philadelphia translocation (vPh). Cytogenetic preparations from 10 CML patients with vPh confirmed by R banding were assayed with dual color dual fusion FISH technique. If only one fusion signal was detected in interphase cells, metaphase cells were observed to determine if there were derivative chromosome 9[der (9)] deletions. Meanwhile, the same cytogenetic preparations were assayed with M-FISH technique. Of the 10 CML patients with vPh, 5 were detected with der (9) deletions by FISH technique. M-FISH technique revealed that besides the chromosome 22, chromosomes 1, 3, 5, 6, 8, 10, 11 and 17 were also involved in the vPh. M-FISH technique also detected the abnormalities which were not found with conventional cytogenetics (CC), including two never reported abnormalities. The combination of CC, FISH and M-FISH technique could refine the genetic diagnosis of CML with vPh.

The Virtual Physiological Human - a European initiative for in silico human modelling -.

Science.gov (United States)

Viceconti, Marco; Clapworthy, Gordon; Van Sint Jan, Serge

2008-12-01

The Virtual Physiological Human (VPH) is an initiative, strongly supported by the European Commission (EC), that seeks to develop an integrated model of human physiology at multiple scales from the whole body through the organ, tissue, cell and molecular levels to the genomic level. VPH had its beginnings in 2005 with informal discussions amongst like-minded scientists which led to the STEP project, a Coordination Action funded by the EC that began in early 2006. The STEP project greatly accelerated the progress of the VPH and proved to be a catalyst for wide-ranging discussions within Europe and for outreach activities designed to develop a broad international approach to the huge scientific and technological challenges involved in this area. This paper provides an overview of the VPH and the developments it has engendered in the rapidly expanding worldwide activities associated with the physiome. It then uses one particular project, the Living Human Project, to illustrate the type of advances that are taking place to further the aims of the VPH and similar initiatives worldwide.

Diseño óptimo de un disipador de calor para luminaria LED mediante moderación modelación computacional

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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.

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...

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...

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.

Performance of Droplet Digital PCR in Non-Invasive Fetal RHD Genotyping - Comparison with a Routine Real-Time PCR Based Approach.

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Iveta Svobodová

Full Text Available Detection and characterization of circulating cell-free fetal DNA (cffDNA from maternal circulation requires an extremely sensitive and precise method due to very low cffDNA concentration. In our study, droplet digital PCR (ddPCR was implemented for fetal RHD genotyping from maternal plasma to compare this new quantification alternative with real-time PCR (qPCR as a golden standard for quantitative analysis of cffDNA. In the first stage of study, a DNA quantification standard was used. Clinical samples, including 10 non-pregnant and 35 pregnant women, were analyzed as a next step. Both methods' performance parameters-standard curve linearity, detection limit and measurement precision-were evaluated. ddPCR in comparison with qPCR has demonstrated sufficient sensitivity for analysing of cffDNA and determination of fetal RhD status from maternal circulation, results of both methods strongly correlated. Despite the more demanding workflow, ddPCR was found to be slightly more precise technology, as evaluated using quantitative standard. Regarding the clinical samples, the precision of both methods equalized with decreasing concentrations of tested DNA samples. In case of cffDNA with very low concentrations, variance parameters of both techniques were comparable. Detected levels of fetal cfDNA in maternal plasma were slightly higher than expected and correlated significantly with gestational age as measured by both methods (ddPCR r = 0.459; qPCR r = 0.438.

Environmental Assessment: Mid-Bay Bridge Connector

Science.gov (United States)

2008-11-01

Speed (mph) AADT VPH Receptor Max 1-Hr Conc (ppm) Max 8-Hr Conc (ppm) Proposed Action 2001 45 17,400 981 Proposed Intersection at SR 20...AADT VPH Receptor Max 1-Hr Conc (ppm) Max 8-Hr Conc (ppm) Alternative C 2001 45 17,400 981 Proposed Intersection at SR 20 5.6 3.3...Volumes Alternative Year Average Speed White Point Road (mph) AADT VPH Receptor Max 1-Hr Conc (ppm) Max 8-Hr Conc (ppm) No-Action 2001

Use of Compound-Specific Stable Isotope Analysis to Distinguish Between Vapor Intrusion and Indoor Sources of VOCs

Science.gov (United States)

2013-11-01

Chlorinated Hydrocarbons, Volatile Organics, TPH (HEM/SGT), CT- Extractable Petroleum Hydrocarbons (ETPH), MA-EPH, MA- VPH . Microbiology Parameters: Total...Toxaphene, CT-Extractable Petroleum Hydrocarbons (ETPH), MA-EPH, MA- VPH , Dicamba, 2,4-D, 2,4,5-T, 2,4,5-TP(Silvex), Dalapon, Volatile Organics (SW 8260...3510C, 3630C, 5030B, ME- DRO, ME-GRO, MA-EPH, MA- VPH .) Solid Waste/Soil (Inorganic Parameters: 9010B, 9012A, 9014A, 9030B, 9040B, 9045C, 6010B

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...

Detection of five potentially periodontal pathogenic bacteria in peri-implant disease: A comparison of PCR and real-time PCR.

Science.gov (United States)

Schmalz, Gerhard; Tsigaras, Sandra; Rinke, Sven; Kottmann, Tanja; Haak, Rainer; Ziebolz, Dirk

2016-07-01

The aim of this study was to compare the microbial analysis methods of polymerase chain reaction (PCR) and real-time PCR (RT-PCR) in terms of detection of five selected potentially periodontal pathogenic bacteria in peri-implant disease. Therefore 45 samples of healthy, mucositis and peri-implantitis (n = 15 each) were assessed according to presence of the following bacteria using PCR (DNA-strip technology) and RT-PCR (fluorescent dye SYBR green-system): Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tanerella forsythia (Tf), and Fusobacterium nucleatum (Fn). There were no significant correlations between the bacterial and disease patterns, so the benefit of using microbiological tests for the diagnosis of peri-implant diseases is questionable. Correlations between the methods were highest for Tf (Kendall's Tau: 0.65, Spearman: 0.78), Fn (0.49, 0.61) and Td (0.49, 0.59). For Aa (0.38, 0.42) and Pg (0.04, 0.04), lower correlation values were detected. Accordingly, conventional semi-quantitative PCR seems to be sufficient for analyzing potentially periodontal pathogenic bacterial species. Copyright © 2016 Elsevier Inc. All rights reserved.

Seguimiento de trayectorias tridimensionales de un quadrotor mediante control PVA

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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

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....

Development and Evaluation of a PCR and Mass Spectroscopy-based (PCR-MS) Method for Quantitative, Type-specific Detection of Human Papillomavirus

Science.gov (United States)

Patel, Divya A.; Shih, Yang-Jen; Newton, Duane W.; Michael, Claire W.; Oeth, Paul A.; Kane, Michael D.; Opipari, Anthony W.; Ruffin, Mack T.; Kalikin, Linda M.; Kurnit, David M.

2010-01-01

Knowledge of the central role of high-risk human papillomavirus (HPV) in cervical carcinogenesis, coupled with an emerging need to monitor the efficacy of newly introduced HPV vaccines, warrant development and evaluation of type-specific, quantitative HPV detection methods. In the present study, a prototype PCR and mass spectroscopy (PCR-MS)-based method to detect and quantitate 13 high-risk HPV types is compared to the Hybrid Capture 2 High Risk HPV DNA test (HC2; Digene Corp., Gaithersburg, MD) in 199 cervical scraping samples and to DNA sequencing in 77 cervical tumor samples. High-risk HPV types were detected in 76/77 (98.7%) cervical tumor samples by PCR-MS. Degenerate and type-specific sequencing confirmed the types detected by PCR-MS. In 199 cervical scraping samples, all 13 HPV types were detected by PCR-MS. Eighteen (14.5%) of 124 cervical scraping samples that were positive for high-risk HPV by HC2 were negative by PCR-MS. In all these cases, degenerate DNA sequencing failed to detect any of the 13 high-risk HPV types. Nearly half (46.7%) of the 75 cervical scraping samples that were negative for high-risk HPV by the HC2 assay were positive by PCR-MS. Type-specific sequencing in a subset of these samples confirmed the HPV type detected by PCR-MS. Quantitative PCR-MS results demonstrated that 11/75 (14.7%) samples contained as much HPV copies/cell as HC2-positive samples. These findings suggest that this prototype PCR-MS assay performs at least as well as HC2 for HPV detection, while offering the additional, unique advantages of type-specific identification and quantitation. Further validation work is underway to define clinically meaningful HPV detection thresholds and to evaluate the potential clinical application of future generations of the PCR-MS assay. PMID:19410602

Development and evaluation of a PCR and mass spectroscopy (PCR-MS)-based method for quantitative, type-specific detection of human papillomavirus.

Science.gov (United States)

Patel, Divya A; Shih, Yang-Jen; Newton, Duane W; Michael, Claire W; Oeth, Paul A; Kane, Michael D; Opipari, Anthony W; Ruffin, Mack T; Kalikin, Linda M; Kurnit, David M

2009-09-01

Knowledge of the central role of high-risk human papillomavirus (HPV) in cervical carcinogenesis, coupled with an emerging need to monitor the efficacy of newly introduced HPV vaccines, warrant development and evaluation of type-specific, quantitative HPV detection methods. In the present study, a prototype PCR and mass spectroscopy (PCR-MS)-based method to detect and quantitate 13 high-risk HPV types is compared to the Hybrid Capture 2 High-Risk HPV DNA test (HC2; Digene Corp., Gaithersburg, MD) in 199 cervical scraping samples and to DNA sequencing in 77 cervical tumor samples. High-risk HPV types were detected in 76/77 (98.7%) cervical tumor samples by PCR-MS. Degenerate and type-specific sequencing confirmed the types detected by PCR-MS. In 199 cervical scraping samples, all 13 HPV types were detected by PCR-MS. Eighteen (14.5%) of 124 cervical scraping samples that were positive for high-risk HPV by HC2 were negative by PCR-MS. In all these cases, degenerate DNA sequencing failed to detect any of the 13 high-risk HPV types. Nearly half (46.7%) of the 75 cervical scraping samples that were negative for high-risk HPV by the HC2 assay were positive by PCR-MS. Type-specific sequencing in a subset of these samples confirmed the HPV type detected by PCR-MS. Quantitative PCR-MS results demonstrated that 11/75 (14.7%) samples contained as much HPV copies/cell as HC2-positive samples. These findings suggest that this prototype PCR-MS assay performs at least as well as HC2 for HPV detection, while offering the additional, unique advantages of type-specific identification and quantitation. Further validation work is underway to define clinically meaningful HPV detection thresholds and to evaluate the potential clinical application of future generations of the PCR-MS assay.

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.

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

Directory of Open Access Journals (Sweden)

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.

Application of real-time PCR (qPCR) for characterization of microbial populations and type of milk in dairy food products.

Science.gov (United States)

Agrimonti, Caterina; Bottari, Benedetta; Sardaro, Maria Luisa Savo; Marmiroli, Nelson

2017-09-08

Dairy foods represent an important sector of the food market for their nutritional qualities and their organoleptic characteristics, which are often linked to tradition and to region. These products are typically protected by labels such as PDO (Protected Designation of Origin) and PGI (Protected Geographical Indication). Real-time PCR (qPCR) is a fundamental tool in "Food Genomics;" a discipline concerned with the residual DNA in food, which, alongside traditional physical and chemical methods, is frequently used to determine product safety, quality and authenticity. Compared to conventional or "end-point" PCR, qPCR incorporates continuous monitoring of reaction progress, thereby enabling quantification of target DNA. This review describes qPCR applications to the analysis of microbiota, and to the identification of the animal species source of milk from which dairy products have been made. These are important aspects for ensuring safety and authenticity. The various applications of qPCR are discussed, as well as advantages and disadvantages in comparison with other analytical methods.

SÍNTESIS DE ÓXIDOS TIPO PEROVSKITA MEDIANTE POLIMERIZACIÓN CON ÁCIDO CÍTRICO Y PROPIÓNICO

Directory of Open Access Journals (Sweden)

Jairo Gómez Cuaspud

2010-03-01

Full Text Available En este trabajo se describe la preparación de la perovskita La0,75Sr0,25Co0,5Fe0,5O3 (LaSrCoFeO, empleando una ruta de química húmeda, mediante la polimerización con ácido cítrico y propiónico, con el propósito de obtener materiales para potenciales aplicaciones como membranas de purificación de oxígeno y como materiales electródicos en celdas de combustible de óxido sólido (SOFC. Para ello, los sólidos se caracterizaron mediante difracción de rayos X (DRX y microscopia electrónica de barrido (SEM, con lo que se obtuvo información sobre la formación y pureza de fases, la morfología, la estructura y las propiedades superficiales de cada sistema, indicando que es posible obtener sólidos con una distribución de grano homogéneo, textura y relieve característicos, en cuyo contexto el método que involucra la polimerización con ácido cítrico mostró los mejores resultados. La composición global se determinó mediante microanálisis de rayos X de energía dispersiva (EDX, y se señaló una buena concordancia entre las composiciones propuestas y obtenidas. La caracterización realizada sugiere la presencia de pequeñas cantidades de carbono y algunos óxidos de lantano, estroncio y cobalto como principales contaminantes, específicamente en la muestra obtenida mediante la polimerización con ácido propiónico.

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.

[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.

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.

Characterization of Freshwater EM Sub Bottom Sediment Properties and Target Responses for Detection of UXO with Ground-Penetrating RADAR (GPR)

Science.gov (United States)

2008-09-01

such that n*= √ε*. We computed phase velocity vph = c/Real(n*). We computed the one-way attenuation rate β (dB m−1) from the imaginary part of the...velocities of propagation at 100 MHz and 1 GHz. At 1 GHz we might expect vph to be controlled by the free, or nearly free value of εshi. The complex...distorted waveform resulted from changes in vph , β, or both across the pulse bandwidth. The small differences in vphmeas between 100 MHz and 1 GHz at

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.

IDENTIFIKASI TIPE HLA KELAS II DENGAN TEKNIK PCR

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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

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.

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...

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.

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...

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.

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.

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.

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.

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

Comparative Evaluation Of Conventional Rt-pcr And Real-time Rt-pcr (rrt-pcr) For Detection Of Avian Metapneumovirus Subtype A [comparação Entre As Técnicas De Rt-pcr Convencional E Rt-pcr Em Tempo Real Para A Detecção Do Metapneumovírus Aviários Subtipo A

OpenAIRE

Ferreira H.L.; Spilki F.R.; dos Santos M.M.A.B.; de Almeida R.S.; Arns C.W.

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...

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.

Principios básicos y aplicación del aprendizaje mediante tareas

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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.

Pronósticos de inflación mediante técnicas bayesianas

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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.

MULTIPLEX SYBR® GREEN-REAL TIME PCR (qPCR ASSAY FOR THE DETECTION AND DIFFERENTIATION OF Bartonella henselae AND Bartonella clarridgeiae IN CATS

Directory of Open Access Journals (Sweden)

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.

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.

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.

Historia natural de la infección por el virus del papiloma humano: una actualización

OpenAIRE

González Martínez, Gerardo; Núñez Troconis, José

2014-01-01

Durante los últimos años, se han sucedido grandes avances en nuestro entendimiento acerca de la biología e historia natural del Virus del Papiloma Humano (VPH). La mayoría de las infecciones por papiloma virus son transmitidas por un contacto cercano bien sea de piel a piel o mucosa a mucosa. La relación sexual con penetración no es un requerimiento para la transmisión del VPH. Las infecciones orales y digitales por VPH ocurren, y existe evidencia de que el contacto digital-genital y genital-...

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.

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.

Have you tried spermine? A rapid and cost-effective method to eliminate dextran sodium sulfate inhibition of PCR and RT-PCR.

Science.gov (United States)

Krych, Łukasz; Kot, Witold; Bendtsen, Katja M B; Hansen, Axel K; Vogensen, Finn K; Nielsen, Dennis S

2018-01-01

The Dextran Sulfate Sodium (DSS) induced colitis mouse model is commonly used to investigate human inflammatory bowel disease (IBD). Nucleic acid extracts originating from these animals are often contaminated with DSS, which is a strong inhibitor of many enzymatic based molecular biology reactions including PCR and reverse-transcription (RT). Methods for removing DSS from nucleic acids extracts exist for RNA, but no effective protocol for DNA or cDNA is currently available. However, spermine has previously been shown to be an effective agent for counteracting DSS inhibition of polynucleotide kinase, which led to the hypothesis, that spermine could be used to counteract DSS inhibition of PCR and RT. We investigated the means of adding spermine in an adequate concentration to PCR based protocols (including qPCR, two-step RT-qPCR, and amplicon sequencing library preparation) to remove DSS inhibition. Within the range up to 0.01g/L, spermine can be added to PCR/qPCR or RT prophylactically without a significant reduction of reaction efficiency. Addition of spermine at the concentration of 0.08g/L can be used to recover qualitative PCR signal inhibited by DSS in concentrations up to 0.32g/L. For optimal quantitative analysis, the concentration of spermine requires fine adjustment. Hence, we present here a simple fluorometric based method for adjusting the concentration of spermine ensuring an optimal efficiency of the reaction exposed to an unknown concentration of DSS. In conclusion, we demonstrate a cost effective and easy method to counteract DSS inhibition in PCR and two-step RT-qPCR. Fixed or fine-tuned concentrations of spermine can be administered depending on the qualitative or quantitative character of the analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

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 ...

The diagnosis of microorganism involved in infective endocarditis (IE by polymerase chain reaction (PCR and real-time PCR: A systematic review

Directory of Open Access Journals (Sweden)

Reza Faraji

2018-02-01

Full Text Available Broad-range bacterial rDNA polymerase chain reaction (PCR followed by sequencing may be identified as the etiology of infective endocarditis (IE from surgically removed valve tissue; therefore, we reviewed the value of molecular testing in identifying organisms' DNA in the studies conducted until 2016. We searched Google Scholar, Scopus, ScienceDirect, Cochrane, PubMed, and Medline electronic databases without any time limitations up to December 2016 for English studies reporting microorganisms involved in infective endocarditis microbiology using PCR and real-time PCR. Most studies were prospective. Eleven out of 12 studies used valve tissue samples and blood cultures while only 1 study used whole blood. Also, 10 studies used the molecular method of PCR while 2 studies used real-time PCR. Most studies used 16S rDNA gene as the target gene. The bacteria were identified as the most common microorganisms involved in infective endocarditis. Streptococcus spp. and Staphylococcus spp. were, by far, the most predominant bacteria detected. In all studies, PCR and real-time PCR identified more pathogens than blood and tissue cultures; moreover, the sensitivity and specificity of PCR and real-time PCR were more than cultures in most of the studies. The highest sensitivity and specificity were 96% and 100%, respectively. The gram positive bacteria were the most frequent cause of infective endocarditis. The molecular methods enjoy a greater sensitivity compared to the conventional blood culture methods; yet, they are applicable only to the valve tissue of the patients undergoing cardiac valve surgery.

The diagnosis of microorganism involved in infective endocarditis (IE) by polymerase chain reaction (PCR) and real-time PCR: A systematic review.

Science.gov (United States)

Faraji, Reza; Behjati-Ardakani, Mostafa; Moshtaghioun, Seyed Mohammad; Kalantar, Seyed Mehdi; Namayandeh, Seyedeh Mahdieh; Soltani, Mohammadhossien; Emami, Mahmood; Zandi, Hengameh; Firoozabadi, Ali Dehghani; Kazeminasab, Mahmood; Ahmadi, Nastaran; Sarebanhassanabadi, Mohammadtaghi

2018-02-01

Broad-range bacterial rDNA polymerase chain reaction (PCR) followed by sequencing may be identified as the etiology of infective endocarditis (IE) from surgically removed valve tissue; therefore, we reviewed the value of molecular testing in identifying organisms' DNA in the studies conducted until 2016. We searched Google Scholar, Scopus, ScienceDirect, Cochrane, PubMed, and Medline electronic databases without any time limitations up to December 2016 for English studies reporting microorganisms involved in infective endocarditis microbiology using PCR and real-time PCR. Most studies were prospective. Eleven out of 12 studies used valve tissue samples and blood cultures while only 1 study used whole blood. Also, 10 studies used the molecular method of PCR while 2 studies used real-time PCR. Most studies used 16S rDNA gene as the target gene. The bacteria were identified as the most common microorganisms involved in infective endocarditis. Streptococcus spp. and Staphylococcus spp. were, by far, the most predominant bacteria detected. In all studies, PCR and real-time PCR identified more pathogens than blood and tissue cultures; moreover, the sensitivity and specificity of PCR and real-time PCR were more than cultures in most of the studies. The highest sensitivity and specificity were 96% and 100%, respectively. The gram positive bacteria were the most frequent cause of infective endocarditis. The molecular methods enjoy a greater sensitivity compared to the conventional blood culture methods; yet, they are applicable only to the valve tissue of the patients undergoing cardiac valve surgery. Copyright © 2017. Published by Elsevier Taiwan.

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.

Comparative evaluation of the nested ITS PCR against the 18S PCR-RFLP in a survey of bovine trypanosomiasis in Kwale County, Kenya.

Science.gov (United States)

Odongo, Steven; Delespaux, Vincent; Ngotho, Maina; Bekkele, Serkalem Mindaye; Magez, Stefan

2016-09-01

We compared the nested internal transcribed spacer (ITS) PCR and the 18S PCR-RFLP (restriction-fragment length polymorphism) pan-trypanosome assays in a cross-sectional survey of bovine trypanosomiasis in 358 cattle in Kwale County, Kenya. The prevalence of trypanosomiasis as determined by the nested ITS PCR was 19.6% (70/358) and by 18S PCR-RFLP was 16.8% (60/358). Of the pathogenic trypanosomes detected, the prevalence of Trypanosoma congolense and Trypanosoma vivax was greater than that of Trypanosoma simiae The nested ITS PCR detected 83 parasite events, whereas the 18S PCR-RFLP detected 64; however, overall frequencies of infections and the parasite events detected did not differ between the assays (χ(2) = 0.8, df = 1, p > 0.05 and χ(2) = 2.5, df = 1, p > 0.05, respectively). The kappa statistic (0.8) showed good agreement between the tests. The nested ITS PCR and the 18S PCR-RFLP had comparable sensitivity, although the nested ITS PCR was better at detecting mixed infections (χ(2) = 5.4, df = 1, p < 0.05). © 2016 The Author(s).

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.

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.

On-lattice agent-based simulation of populations of cells within the open-source Chaste framework

KAUST Repository

Figueredo, G. P.; Joshi, T. V.; Osborne, J. M.; Byrne, H. M.; Owen, M. R.

2013-01-01

been implemented and are available for download as part of the latest public release of Chaste (Cancer, Heart and Soft Tissue Environment; http://www.cs.ox.ac.uk/chaste/), part of the VPH Toolkit (http://toolkit.vph-noe.eu/). The environment

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.

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.

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...

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.

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.

Cutaneous and visceral leishmaniasis co-infection in dogs from Rio de Janeiro, Brazil: evaluation by specific PCR and RFLP-PCR assays

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Marize Quinhones Pires

2014-04-01

Full Text Available Introduction During a diagnostic evaluation of canine visceral leishmaniasis (VL, two of seventeen dogs were found to be co-infected by Leishmania (Viannia braziliensis and Leishmania (Leishmania chagasi. Methods Specific polymerase chain reaction (PCR and restriction fragment length polymorphism-PCR (RFLP-PCR assays were performed. Results PCR assays for Leishmania subgenus identification followed by RFLP-PCR analysis in biopsies from cutaneous lesions and the spleen confirmed the presence of Leishmania (Viannia braziliensis and Leishmania (Leishmania chagasi in those fragments. Conclusions This report reinforces the importance of using serological and molecular techniques in the epidemiological surveillance of canine populations in endemic areas in which both diseases are known to co-exist. In such cases, a reassessment of the control measures is required.

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)

Development of Quantitative Competitive PCR and Absolute Based Real-Time PCR Assays for Quantification of The Butyrate Producing Bacterium: Butyrivibrio fibrisolvens

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Mojtaba Tahmoorespur

2016-04-01

Full Text Available Introduction Butyrivibrio fibrisolvens strains are presently recognized as the major butyrate-producing bacteria found in the rumen and digestive track of many animals and also in the human gut. In this study we reported the development of two DNA based techniques, quantitative competitive (QC PCR and absolute based Real-Time PCR, for enumerating Butyrivibrio fibrisolvens strains. Despite the recent introduction of real-time PCR method for the rapid quantification of the target DNA sequences, use of quantitative competitive PCR (QC-PCR technique continues to play an important role in nucleic acid quantification since it is more cost effective. The procedure relies on the co-amplification of the sequence of interest with a serially diluted synthetic DNA fragment of the known concentration (competitor, using the single set primers. A real-time polymerase chain reaction is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR. It monitors the amplification of a targeted DNA molecule during the PCR. Materials and Methods At first reported species-specific primers targeting the 16S rDNA region of the bacterium Butyrivibrio fibrisolvens were used for amplifying a 213 bp fragment. A DNA competitor differing by 50 bp in length from the 213 bp fragment was constructed and cloned into pTZ57R/T vector. The competitor was quantified by NanoDrop spectrophotometer and serially diluted and co-amplified by PCR with total extracted DNA from rumen fluid samples. PCR products were quantified by photographing agarose gels and analyzed with Image J software and the amount of amplified target DNA was log plotted against the amount of amplified competitor. Coefficient of determination (R2 was used as a criterion of methodology precision. For developing the Real-time PCR technique, the 213 bp fragment was amplified and cloned into pTZ57R/T was used to draw a standard curve. Results and Discussion The specific primers of Butyrivibrio

Curvas de fusión de regiones genómicas específicas: una herramienta prometedora para el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia.

Science.gov (United States)

Marin, Johana; Urrea, Daniel; Muskus, Carlos; Echeverry, María Clara; Mejía, Ana María; Triana, Omar

2017-12-01

Introducción. La leishmaniasis cutánea es una enfermedad causada por parásitos del género Leishmania que tiene gran incidencia en Colombia. El diagnóstico y la identificación de la especie infecciosa son factores críticos en el momento de escoger e iniciar el tratamiento. Actualmente, los métodos de diagnóstico y tipificación requieren procedimientos complejos, por lo que es necesario validar nuevos marcadores moleculares y métodos que simplifiquen el proceso.Objetivo. Desarrollar una herramienta basada en la reacción en cadena de la polimerasa (PCR) con curvas de fusión (High Resolution Melting; PCR-HRM) para el diagnóstico y tipificación de las tres especies de Leishmania de importancia epidemiológica en casos de leishmaniasis cutánea en Colombia.Materiales y métodos. Los genomas de Leishmania panamensis, L. braziliensis y L. guyanensis se compararon mediante métodos bioinformáticos. Las regiones específicas de especie identificadas se validaron mediante PCR. Para los marcadores seleccionados se diseñó una PCR-HRM y se estimaron algunos parámetros de validez y seguridad usando aislamientos de pacientes colombianos caracterizados previamente mediante PCR y análisis de polimorfismos en la longitud de los fragmentos de restricción (Restriction Fragment Length Polymorphism - RFLP; PCR-RFLP) del gen hsp70.Resultados. El análisis genómico comparativo mostró 24 regiones específicas de especie. Sin embargo, la validación mediante PCR solo identificó un marcador específico para cada especie de Leishmania. Los otros marcadores mostraron amplificación cruzada. El límite de detección para los tres marcadores seleccionados fue de un parásito, mientras que la sensibilidad, la especificidad, el valor predictivo positivo y el negativo fueron de 91,4, 100, 100 y 75 %, respectivamente.Conclusiones. Las tres regiones seleccionadas pueden emplearse como marcadores moleculares en el diagnóstico y tipificación de las especies causantes de la

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.

Caracterización espacial de PM10 en la ciudad de Medellín mediante modelos geoestadísticos

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Libardo Antonio Londoño Ciro

2015-12-01

Full Text Available En este artículo se presenta un modelo geoestadístico para caracterizar espacialmente el comportamiento del contaminante PM10 en la ciudad de Medellín Colombia. Los datos se han tomado de nueve sitios de monitoreo en valor promedio mensual (µg/m3 durante el periodo enero 2003 a diciembre 2007. Se evaluaron diferentes modelos mediante pruebas de validación cruzada. El mejor modelo es el j-bessel. Se calculan los parámetros del modelo mediante pruebas ANOVA para agrupaciones trimestrales. Con Kriging ordinario y sistemas de información geográfica, se obtienen mapas de caracterización espacial del contaminante.

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 ...

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...

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...

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...

Two-temperature LATE-PCR endpoint genotyping

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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

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.

Detection of Histoplasma capsulatum from clinical specimens by cycling probe-based real-time PCR and nested real-time PCR.

Science.gov (United States)

Muraosa, Yasunori; Toyotome, Takahito; Yahiro, Maki; Watanabe, Akira; Shikanai-Yasuda, Maria Aparecida; Kamei, Katsuhiko

2016-05-01

We developed new cycling probe-based real-time PCR and nested real-time PCR assays for the detection of Histoplasma capsulatum that were designed to detect the gene encoding N-acetylated α-linked acidic dipeptidase (NAALADase), which we previously identified as an H. capsulatum antigen reacting with sera from patients with histoplasmosis. Both assays specifically detected the DNAs of all H. capsulatum strains but not those of other fungi or human DNA. The limited of detection (LOD) of the real-time PCR assay was 10 DNA copies when using 10-fold serial dilutions of the standard plasmid DNA and 50 DNA copies when using human serum spiked with standard plasmid DNA. The nested real-time PCR improved the LOD to 5 DNA copies when using human serum spiked with standard plasmid DNA, which represents a 10-fold higher than that observed with the real-time PCR assay. To assess the ability of the two assays to diagnose histoplasmosis, we analyzed a small number of clinical specimens collected from five patients with histoplasmosis, such as sera (n = 4), formalin-fixed paraffin-embedded (FFPE) tissue (n = 4), and bronchoalveolar lavage fluid (BALF) (n = 1). Although clinical sensitivity of the real-time PCR assay was insufficiently sensitive (33%), the nested real-time PCR assay increased the clinical sensitivity (77%), suggesting it has a potential to be a useful method for detecting H. capsulatum DNA in clinical specimens. © 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.

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.

Lqr Robusto Mediante Incertidumbre Acotada En Los Datos

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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

Fusion primer and nested integrated PCR (FPNI-PCR: a new high-efficiency strategy for rapid chromosome walking or flanking sequence cloning

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Wang Zhen

2011-11-01

Full Text Available Abstract Background The advent of genomics-based technologies has revolutionized many fields of biological enquiry. However, chromosome walking or flanking sequence cloning is still a necessary and important procedure to determining gene structure. Such methods are used to identify T-DNA insertion sites and so are especially relevant for organisms where large T-DNA insertion libraries have been created, such as rice and Arabidopsis. The currently available methods for flanking sequence cloning, including the popular TAIL-PCR technique, are relatively laborious and slow. Results Here, we report a simple and effective fusion primer and nested integrated PCR method (FPNI-PCR for the identification and cloning of unknown genomic regions flanked known sequences. In brief, a set of universal primers was designed that consisted of various 15-16 base arbitrary degenerate oligonucleotides. These arbitrary degenerate primers were fused to the 3' end of an adaptor oligonucleotide which provided a known sequence without degenerate nucleotides, thereby forming the fusion primers (FPs. These fusion primers are employed in the first step of an integrated nested PCR strategy which defines the overall FPNI-PCR protocol. In order to demonstrate the efficacy of this novel strategy, we have successfully used it to isolate multiple genomic sequences namely, 21 orthologs of genes in various species of Rosaceace, 4 MYB genes of Rosa rugosa, 3 promoters of transcription factors of Petunia hybrida, and 4 flanking sequences of T-DNA insertion sites in transgenic tobacco lines and 6 specific genes from sequenced genome of rice and Arabidopsis. Conclusions The successful amplification of target products through FPNI-PCR verified that this novel strategy is an effective, low cost and simple procedure. Furthermore, FPNI-PCR represents a more sensitive, rapid and accurate technique than the established TAIL-PCR and hiTAIL-PCR procedures.

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

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.

Trypanosoma rangeli: RAPD-PCR and LSSP-PCR analyses of isolates from southeast Brazil and Colombia and their relation with KPI minicircles.

Science.gov (United States)

Marquez, D S; Ramírez, L E; Moreno, J; Pedrosa, A L; Lages-Silva, E

2007-09-01

This study presents the first genetic characterization of five Trypanosoma rangeli isolates from Minas Gerais, in the southeast of Brazil and their comparison with Colombian populations by minicircle classification, RAPD-PCR and LSSP-PCR analyses. Our results demonstrated a homogenous T. rangeli population circulating among Didelphis albiventris as reservoir host in Brazil while heterogeneous populations were found in different regions of Colombia. KP1(+) minicircles were found in 100% isolates from Brazil and in 36.4% of the Colombian samples, whereas the KP2 and KP3 minicircles were detected in both groups. RAPD-PCR and LSSP-PCR profiles revealed a polymorphism within KP1(+) and KP1(-) T. rangeli populations and allowed the division of T. rangeli in two branches. The Brazilian KP1(+) isolates were more homogenous than the KP1(+) isolates from Colombia. The RAPD-PCR were entirely consistent with the distribution of KP1 minicircles while those obtained by LSSP-PCR were associated in 88.9% and 71.4% with KP1(+) and KP1(-) populations, respectively.

Comparison of Direct Sequencing, Real-Time PCR-High Resolution Melt (PCR-HRM) and PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis for Genotyping of Common Thiopurine Intolerant Variant Alleles NUDT15 c.415C>T and TPMT c.719A>G (TPMT*3C).

Science.gov (United States)

Fong, Wai-Ying; Ho, Chi-Chun; Poon, Wing-Tat

2017-05-12

Thiopurine intolerance and treatment-related toxicity, such as fatal myelosuppression, is related to non-function genetic variants encoding thiopurine S-methyltransferase (TPMT) and Nudix hydrolase 15 (NUDT15). Genetic testing of the common variants NUDT15:NM_018283.2:c.415C>T (Arg139Cys, dbSNP rs116855232 T allele) and TPMT: NM_000367.4:c.719A>G (TPMT*3C, dbSNP rs1142345 G allele) in East Asians including Chinese can potentially prevent treatment-related complications. Two complementary genotyping approaches, real-time PCR-high resolution melt (PCR-HRM) and PCR-restriction fragment length morphism (PCR-RFLP) analysis were evaluated using conventional PCR and Sanger sequencing genotyping as the gold standard. Sixty patient samples were tested, revealing seven patients (11.7%) heterozygous for NUDT15 c.415C>T, one patient homozygous for the variant and one patient heterozygous for the TPMT*3C non-function allele. No patient was found to harbor both variants. In total, nine out of 60 (15%) patients tested had genotypic evidence of thiopurine intolerance, which may require dosage adjustment or alternative medication should they be started on azathioprine, mercaptopurine or thioguanine. The two newly developed assays were more efficient and showed complete concordance (60/60, 100%) compared to the Sanger sequencing results. Accurate and cost-effective genotyping assays by real-time PCR-HRM and PCR-RFLP for NUDT15 c.415C>T and TPMT*3C were successfully developed. Further studies may establish their roles in genotype-informed clinical decision-making in the prevention of morbidity and mortality due to thiopurine intolerance.

Introduction on Using the FastPCR Software and the Related Java Web Tools for PCR and Oligonucleotide Assembly and Analysis.

Science.gov (United States)

Kalendar, Ruslan; Tselykh, Timofey V; Khassenov, Bekbolat; Ramanculov, Erlan M

2017-01-01

This chapter introduces the FastPCR software as an integrated tool environment for PCR primer and probe design, which predicts properties of oligonucleotides based on experimental studies of the PCR efficiency. The software provides comprehensive facilities for designing primers for most PCR applications and their combinations. These include the standard PCR as well as the multiplex, long-distance, inverse, real-time, group-specific, unique, overlap extension PCR for multi-fragments assembling cloning and loop-mediated isothermal amplification (LAMP). It also contains a built-in program to design oligonucleotide sets both for long sequence assembly by ligase chain reaction and for design of amplicons that tile across a region(s) of interest. The software calculates the melting temperature for the standard and degenerate oligonucleotides including locked nucleic acid (LNA) and other modifications. It also provides analyses for a set of primers with the prediction of oligonucleotide properties, dimer and G/C-quadruplex detection, linguistic complexity as well as a primer dilution and resuspension calculator. The program consists of various bioinformatical tools for analysis of sequences with the GC or AT skew, CG% and GA% content, and the purine-pyrimidine skew. It also analyzes the linguistic sequence complexity and performs generation of random DNA sequence as well as restriction endonucleases analysis. The program allows to find or create restriction enzyme recognition sites for coding sequences and supports the clustering of sequences. It performs efficient and complete detection of various repeat types with visual display. The FastPCR software allows the sequence file batch processing that is essential for automation. The program is available for download at http://primerdigital.com/fastpcr.html , and its online version is located at http://primerdigital.com/tools/pcr.html .

Bulimia nerviosa

Science.gov (United States)

... Virus del papiloma humano (VPH) Infertilidad Síndrome de ovario poliquístico (SOP) Infecciones de transmisión sexual (ITS) Fibromas ... Virus del papiloma humano (VPH) Infertilidad Síndrome de ovario poliquístico (SOP) Infecciones de transmisión sexual (ITS) Fibromas ...

A Compact, Pi-Mode Extraction Scheme for the Axial B-Field Recirculating Planar Magnetron

Science.gov (United States)

2012-07-23

Figure 4). Thus, in a planar magnetron, the minimum phase velocity, vph , to stay above cutoff in the rectangular waveguide is ℎ = ...as magnetrons, electrons must be accelerated such that they are in synchronism with the phase velocity, vph , of the electromagnetic wave for an

Environmental Assessment, SR 123 (Roger J. Clary Highway) from North of the Intersection of SR 123 and SR 85S to SR 85N, Okaloosa County, Eglin Air Force Base, Florida

Science.gov (United States)

2012-05-01

mph) SRS 123 / SR 85 Traffic Volumes Receptor Max 1-Hr CO Conc (ppm) Max 8-Hr CO Conc (ppm) AADT VPH Alternative 2 (East Shift) and...8.4 5.0 AADT: Annual Average Daily Traffic. VPH : Vehicles per Hour Environmental Consequences Air Quality Impacts

TRANSVERSE MODES IN PHASE-CONJUGATION RESONATORS (PCR) WITH FINITE APERTURES (Ⅱ)——FUNDAMENTAL PROPERTIES OF THE TRANSVERSE MODES IN PCR

Institute of Scientific and Technical Information of China (English)

李先枢; 徐家进; 高燕球

1990-01-01

Based on the first part of this paper (Science in China, 33(1990), 982—995), further research has been done on quasi-equivalence relation and asymmetrical character of axisymmetrical phase-conjugatlon resonator(PCR). A series of calculations for axisymmetrical PCR(hundreds of transverse modes in 66 axisymmetrical PCRs) have been carried out, and the results are compared with those of corresponding conventional laser resonators. Fundamental properties of the transverse modes (TEMs) in PCR are summarized. This makes possible a rough estimation of the properties of various TEMs in these simple PCR, including different geometrical structures.

Tipificación molecular del Virus Papiloma Humano en mujeres con lesiones cervicales de alto grado y cáncer de cérvix atentidas en el área de colposcopía en consulta externa del ION Solca Guayaquil, periodo 2013 - 2014.

OpenAIRE

Morocho Molina, Nancy Jannet

2015-01-01

El Virus del Papiloma Humano (VPH) está asociado directamente con el desarrollo del cáncer de cérvix. Objetivo: tipificación y asociación de VPH de alto riesgo en pacientes con LIEAG y cáncer de cérvix que acudieron a la consulta externa del ION SOLCA-GUAYAQUIL, durante el período 2013- 2014. Metodología: el presente estudio fue de tipo descriptivo, observacional y analítico, los exámenes utilizados fueron el papanicolaou, colposcopia, biopsia de cérvix y tipificación viral del VPH medi...

Quantitative analysis of food and feed samples with droplet digital PCR.

Directory of Open Access Journals (Sweden)

Dany Morisset

Full Text Available In this study, the applicability of droplet digital PCR (ddPCR for routine analysis in food and feed samples was demonstrated with the quantification of genetically modified organisms (GMOs. Real-time quantitative polymerase chain reaction (qPCR is currently used for quantitative molecular analysis of the presence of GMOs in products. However, its use is limited for detecting and quantifying very small numbers of DNA targets, as in some complex food and feed matrices. Using ddPCR duplex assay, we have measured the absolute numbers of MON810 transgene and hmg maize reference gene copies in DNA samples. Key performance parameters of the assay were determined. The ddPCR system is shown to offer precise absolute and relative quantification of targets, without the need for calibration curves. The sensitivity (five target DNA copies of the ddPCR assay compares well with those of individual qPCR assays and of the chamber digital PCR (cdPCR approach. It offers a dynamic range over four orders of magnitude, greater than that of cdPCR. Moreover, when compared to qPCR, the ddPCR assay showed better repeatability at low target concentrations and a greater tolerance to inhibitors. Finally, ddPCR throughput and cost are advantageous relative to those of qPCR for routine GMO quantification. It is thus concluded that ddPCR technology can be applied for routine quantification of GMOs, or any other domain where quantitative analysis of food and feed samples is needed.

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

The EuroPhysiome, STEP and a roadmap for the virtual physiological human.

Science.gov (United States)

Fenner, J W; Brook, B; Clapworthy, G; Coveney, P V; Feipel, V; Gregersen, H; Hose, D R; Kohl, P; Lawford, P; McCormack, K M; Pinney, D; Thomas, S R; Van Sint Jan, S; Waters, S; Viceconti, M

2008-09-13

Biomedical science and its allied disciplines are entering a new era in which computational methods and technologies are poised to play a prevalent role in supporting collaborative investigation of the human body. Within Europe, this has its focus in the virtual physiological human (VPH), which is an evolving entity that has emerged from the EuroPhysiome initiative and the strategy for the EuroPhysiome (STEP) consortium. The VPH is intended to be a solution to common infrastructure needs for physiome projects across the globe, providing a unifying architecture that facilitates integration and prediction, ultimately creating a framework capable of describing Homo sapiens in silico. The routine reliance of the biomedical industry, biomedical research and clinical practice on information technology (IT) highlights the importance of a tailor-made and robust IT infrastructure, but numerous challenges need to be addressed if the VPH is to become a mature technological reality. Appropriate investment will reap considerable rewards, since it is anticipated that the VPH will influence all sectors of society, with implications predominantly for improved healthcare, improved competitiveness in industry and greater understanding of (patho)physiological processes. This paper considers issues pertinent to the development of the VPH, highlighted by the work of the STEP consortium.

In vitro culture, PCR , and nested PCR for the detection of Theileria equi in horses submitted to exercise Cultivo in vitro, PCR e nested PCR na detecção de Theileria equi em eqüinos submetidos a exercícios

Directory of Open Access Journals (Sweden)

C.D. Baldani

2008-06-01

Full Text Available This study compared the usefulness of in vitro culture, PCR, and nested PCR for the diagnosis of Theileria equi in horses submitted to stress during exercise. Blood samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken prior to and after exercise. The animals were divided into two experimental groups: 30-day training schedule (G1 and 90-day training schedule (G2. Statistical analysis was performed using a chi-square test and kappa statistic was used in order to assess agreement. No significant difference was observed between samples collected at resting or after exercise. In G1, merozoites of T. equi were detected in the blood smears of four horses before in vitro culture, whereas 14 samples were positive, confirmed by culture. In G2, five and 11 horses were positive before and after culture, respectively. No PCR amplified product was observed in any of the tested animals although the PCR system based on the 16S rRNA gene of T. equi detected DNA in blood with an equivalent 8x10-5% parasitaemia. The nested PCR based on the T. equi merozoite antigen gene (EMA-1 allowed the visualization of amplified products in all the horses. Therefore, nested PCR should be considered as a means of detection of sub-clinical T. equi infections and in vitro culture could be used as a complement to other methods of diagnosis.Comparou-se a utilização do cultivo in vitro, PCR e nested PCR no diagnóstico de Theileria equi em eqüinos submetidos ao estresse induzido por exercícios. Amostras de sangue foram obtidas de 15 eqüinos submetidos a treinamento em esteira rolante de alto desempenho, sendo as amostras colhidas antes e após os exercícios. Os animais foram divididos em dois grupos experimentais: 30 dias de treinamento (G1 e 90 dias de treinamento (G2. O teste do qui-quadrado foi empregado para as análises estatísticas e o índice kappa utilizado para avaliar a concordância. Não houve diferen

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.

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.

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.

Influence of DNA extraction methods, PCR inhibitors and quantification methods on real-time PCR assay of biotechnology-derived traits.

Science.gov (United States)

Demeke, Tigst; Jenkins, G Ronald

2010-03-01

Biotechnology-derived varieties of canola, cotton, corn and soybean are being grown in the USA, Canada and other predominantly grain exporting countries. Although the amount of farmland devoted to production of biotechnology-derived crops continues to increase, lingering concerns that unintended consequences may occur provide the EU and most grain-importing countries with justification to regulate these crops. Legislation in the EU requires traceability of grains/oilseeds, food and feed products, and labelling, when a threshold level of 0.9% w/w of genetically engineered trait is demonstrated to be present in an analytical sample. The GE content is routinely determined by quantitative PCR (qPCR) and plant genomic DNA provides the template for the initial steps in this process. A plethora of DNA extraction methods exist for qPCR applications. Implementing standardized methods for detection of genetically engineered traits is necessary to facilitate grain marketing. The International Organization for Standardization draft standard 21571 identifies detergent-based methods and commercially available kits that are widely used for DNA extraction, but also indicates that adaptations may be necessary depending upon the sample matrix. This review assesses advantages and disadvantages of various commercially available DNA extraction kits, as well as modifications to published cetyltrimethylammonium bromide methods. Inhibitors are a major obstacle for efficient amplification in qPCR. The types of PCR inhibitors and techniques to minimize inhibition are discussed. Finally, accurate quantification of DNA for applications in qPCR is not trivial. Many confounders contribute to differences in analytical measurements when a particular DNA quantification method is applied and different methods do not always provide concordant results on the same DNA sample. How these differences impact measurement uncertainty in qPCR is considered.

Rapid and sensitive detection of Feline immunodeficiency virus using an insulated isothermal PCR-based assay with a point-of-need PCR detection platform.

Science.gov (United States)

Wilkes, Rebecca Penrose; Kania, Stephen A; Tsai, Yun-Long; Lee, Pei-Yu Alison; Chang, Hsiu-Hui; Ma, Li-Juan; Chang, Hsiao-Fen Grace; Wang, Hwa-Tang Thomas

2015-07-01

Feline immunodeficiency virus (FIV) is an important infectious agent of cats. Clinical syndromes resulting from FIV infection include immunodeficiency, opportunistic infections, and neoplasia. In our study, a 5' long terminal repeat/gag region-based reverse transcription insulated isothermal polymerase chain reaction (RT-iiPCR) was developed to amplify all known FIV strains to facilitate point-of-need FIV diagnosis. The RT-iiPCR method was applied in a point-of-need PCR detection platform--a field-deployable device capable of generating automatically interpreted RT-iiPCR results from nucleic acids within 1 hr. Limit of detection 95% of FIV RT-iiPCR was calculated to be 95 copies standard in vitro transcription RNA per reaction. Endpoint dilution studies with serial dilutions of an ATCC FIV type strain showed that the sensitivity of lyophilized FIV RT-iiPCR reagent was comparable to that of a reference nested PCR. The established reaction did not amplify any nontargeted feline pathogens, including Felid herpesvirus 1, feline coronavirus, Feline calicivirus, Feline leukemia virus, Mycoplasma haemofelis, and Chlamydophila felis. Based on analysis of 76 clinical samples (including blood and bone marrow) with the FIV RT-iiPCR, test sensitivity was 97.78% (44/45), specificity was 100.00% (31/31), and agreement was 98.65% (75/76), determined against a reference nested-PCR assay. A kappa value of 0.97 indicated excellent correlation between these 2 methods. The lyophilized FIV RT-iiPCR reagent, deployed on a user-friendly portable device, has potential utility for rapid and easy point-of-need detection of FIV in cats. © 2015 The Author(s).

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.

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...

Optimized PCR with sequence specific primers (PCR-SSP for fast and efficient determination of Interleukin-6 Promoter -597/-572/-174Haplotypes

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Bugert Peter

2009-12-01

Full Text Available Abstract Background Interleukin-6 (IL-6 promoter polymorphisms at positions -597(G→A, -572(G→C and -174(G→C were shown to have a clinical impact on different major diseases. At present PCR-SSP protocols for IL-6 -597/-572/-174haplotyping are elaborate and require large amounts of genomic DNA. Findings We describe an improved typing technique requiring a decreased number of PCR-reactions and a reduced PCR-runtime due to optimized PCR-conditions. Conclusion This enables a fast and efficient determination of IL-6 -597/-572/-174haplotypes in clinical diagnosis and further evaluation of IL-6 promoter polymorphisms in larger patient cohorts.

Knowledge environments representing molecular entities for the virtual physiological human.

Science.gov (United States)

Hofmann-Apitius, Martin; Fluck, Juliane; Furlong, Laura; Fornes, Oriol; Kolárik, Corinna; Hanser, Susanne; Boeker, Martin; Schulz, Stefan; Sanz, Ferran; Klinger, Roman; Mevissen, Theo; Gattermayer, Tobias; Oliva, Baldo; Friedrich, Christoph M

2008-09-13

In essence, the virtual physiological human (VPH) is a multiscale representation of human physiology spanning from the molecular level via cellular processes and multicellular organization of tissues to complex organ function. The different scales of the VPH deal with different entities, relationships and processes, and in consequence the models used to describe and simulate biological functions vary significantly. Here, we describe methods and strategies to generate knowledge environments representing molecular entities that can be used for modelling the molecular scale of the VPH. Our strategy to generate knowledge environments representing molecular entities is based on the combination of information extraction from scientific text and the integration of information from biomolecular databases. We introduce @neuLink, a first prototype of an automatically generated, disease-specific knowledge environment combining biomolecular, chemical, genetic and medical information. Finally, we provide a perspective for the future implementation and use of knowledge environments representing molecular entities for the VPH.

Vertical parasagittal hemispherotomy for Sturge-Weber syndrome in early infancy: case report and literature review.

Science.gov (United States)

Liu, Xiangyu; Otsuki, Taisuke; Takahashi, Akio; Kaido, Takanobu

2016-01-01

The authors here present a rare case of a 3-month-old infant with unilateral Sturge-Weber syndrome (SWS) who had excellent seizure control and no aggravation of previous existed neurological deficits after vertical parasagittal hemispherotomy (VPH). To our knowledge, this patient with SWS was the youngest one who received VPH. The use of VPH results in a successful treatment of intractable epilepsy in a patient with seizure onset in early infancy. At follow-up, the patient's neurodevelopmental status has been improved since the surgery. It is generally accepted that early-onset seizures in children with SWS are associated with worse neurological and developmental outcomes. However, when surgical treatment should be considered and how it should be performed remain a longstanding controversy. We promote early surgery in children with SWS and early-onset epilepsy. We suggest that VPH may be a useful adjuvant in the management of SWS with refractory epilepsy in early infancy and this procedure carries low neurological risk.

PH responsive self-assembly of cucurbit[7]urils and polystyrene-block- polyvinylpyridine micelles for hydrophobic drug delivery

KAUST Repository

Moosa, Basem; Mashat, A.; Li, W.; Fhayli, K.; Khashab, Niveen M.

2013-01-01

Polystyrene-block-polyvinylpyridine (PS-b-P4VP) polypseudorotaxanes with cucurbit[7]urils (CB[7]) were prepared from water soluble PS-b-P4VPH+ polymer and CB[7] in aqueous solution at room temperature. At acidic and neutral pH, the pyridinium block of PS-b-P4VP is protonated (PS-b-P4VPH +) pushing CB[7] to preferably host the P4VP block. At basic pH (pH 8), P4VP is not charged and thus is not able to strongly complex CB[7]. This phenomenon was verified further by monitoring the release of pyrene, a hydrophobic cargo model, from a PS-b-P4VPH+/CB[7] micellar membrane. Release study of UV active pyrene from the membrane at different pH values revealed that the system is only operational under basic conditions and that the host-guest interaction of CB[7] with P4VPH+ significantly slows down cargo release.

pH Responsive Self-Assembly of Cucurbit[7]urils and Polystyrene-Block-Polyvinylpyridine Micelles for Hydrophobic Drug Delivery

Directory of Open Access Journals (Sweden)

Basem A. Moosa

2013-01-01

Full Text Available Polystyrene-block-polyvinylpyridine (PS-b-P4VP polypseudorotaxanes with cucurbit[7]urils (CB[7] were prepared from water soluble PS-b-P4VPH+ polymer and CB[7] in aqueous solution at room temperature. At acidic and neutral pH, the pyridinium block of PS-b-P4VP is protonated (PS-b-P4VPH+ pushing CB[7] to preferably host the P4VP block. At basic pH (pH 8, P4VP is not charged and thus is not able to strongly complex CB[7]. This phenomenon was verified further by monitoring the release of pyrene, a hydrophobic cargo model, from a PS-b-P4VPH+/CB[7] micellar membrane. Release study of UV active pyrene from the membrane at different pH values revealed that the system is only operational under basic conditions and that the host-guest interaction of CB[7] with P4VPH+ significantly slows down cargo release.

Detección molecular del virus papiloma humano de alto riesgo oncogénico en muestras cervicales. Laboratorio Central de Salud Pública. Primeros Resultados

Directory of Open Access Journals (Sweden)

Maria Liz Bobadilla

2015-04-01

Full Text Available El cáncer de cuello uterino es la primera causa de muerte por cáncer en mujeres en países en vías de desarrollo, con una tasa de incidencia de 34,2 por 100.000 mujeres y de mortalidad de 15,7 por 100.000 mujeres en Paraguay. La sensibilidad de la citología está entre 30-60%, mientras que la de la detección molecular del Virus Papiloma Humano (VPH en muestras cervicales, es mayor al 90% para detectar neoplasia intraepitelial cervical de grado 2 (CIN II o más. El objetivo de este trabajo fue describir la frecuencia de detección de VPH de alto riesgo (AR y su distribución por edad en mujeres que concurrieron al Hospital San Pablo, de mayo a agosto de 2.013. Se estudiaron 170 muestras cervicales de pacientes que accedieron a participar firmando un consentimiento informado. Se utilizó el sistema Cobas 4800 HPV Test (Roche que detecta los VPH-AR 16 y 18, y un pool de 10 VPH-AR (31,33,35,39,45,51,52,56,58,59 y dos de “probable” alto riesgo (66,68. La frecuencia de infección por VPH-AR fue del 16%, la infección decrecía con la edad y el mayor número de casos apareció en mujeres menores de 30 años. El VPH-16 fue encontrado en todos los grupos de edades. Este es el primer reporte de la detección de ADN de VPH-AR en el LCSP, y se muestra que la prevención y control del cáncer cérvico-uterino es una prioridad de salud pública en el país por la gran carga de la enfermedad evidenciada por su alta incidencia y mortalidad.

Detección de virus papiloma humano en la prevención del cáncer cérvico-uterino

Directory of Open Access Journals (Sweden)

María Alejandra Picconi

2013-12-01

Full Text Available El cáncer cérvico-uterino (CCU, que está fuertemente asociado a la infección por virus papiloma humano de alto riesgo (VPH-AR, sigue siendo un problema de salud pública en Latinoamérica. El uso de la citología para la detección de lesiones pre-cancerosas no ha tenido mayor impacto en las tasas de incidencia y mortalidad del CCU, que aún se mantienen altas en la región. La disponibilidad de nuevas técnicas de tamizaje para la detección de lesiones pre-cancerosas y de vacunas altamente eficaces que previenen casi todas las lesiones relacionadas con los VPH-AR de alto potencial oncogénico VPH 16 y 18, en mujeres no expuestas previamente al virus brindan una gran oportunidad para la prevención del CCU. La detección de VPH-AR representa actualmente un valioso componente de las guías clínicas para el tamizaje, manejo y tratamiento del CCU y sus lesiones precursoras. Se han desarrollado estrategias metodológicas que detectan un amplio espectro de tipos de VPH-AR; sin embargo, solo un pequeño subgrupo de ellas ha documentado la validación clínica para cualquiera de las indicaciones habituales de la detección de estos virus. Las pruebas de VPH que no estén validadas y que no hayan demostrado confiabilidad, reproducibilidad y exactitud no deben ser usadas en el manejo clínico. Una vez incorporada una prueba de VPH en el laboratorio, es esencial que el procedimiento completo sea sometido a un continuo y riguroso control de calidad para evitar prácticas subóptimas, potencialmente dañinas. Este artículo discute los recientes progresos y el estado actual de estos métodos.

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...

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...

Quantitative threefold allele-specific PCR (QuanTAS-PCR) for highly sensitive JAK2 V617F mutant allele detection

International Nuclear Information System (INIS)

Zapparoli, Giada V; Jorissen, Robert N; Hewitt, Chelsee A; McBean, Michelle; Westerman, David A; Dobrovic, Alexander

2013-01-01

The JAK2 V617F mutation is the most frequent somatic change in myeloproliferative neoplasms, making it an important tumour-specific marker for diagnostic purposes and for the detection of minimal residual disease. Sensitive quantitative assays are required for both applications, particularly for the monitoring of minimal residual disease, which requires not only high sensitivity but also very high specificity. We developed a highly sensitive probe-free quantitative mutant-allele detection method, Quantitative Threefold Allele-Specific PCR (QuanTAS-PCR), that is performed in a closed-tube system, thus eliminating the manipulation of PCR products. QuantTAS-PCR uses a threefold approach to ensure allele-specific amplification of the mutant sequence: (i) a mutant allele-specific primer, (ii) a 3′dideoxy blocker to suppress false-positive amplification from the wild-type template and (iii) a PCR specificity enhancer, also to suppress false-positive amplification from the wild-type template. Mutant alleles were quantified relative to exon 9 of JAK2. We showed that the addition of the 3′dideoxy blocker suppressed but did not eliminate false-positive amplification from the wild-type template. However, the addition of the PCR specificity enhancer near eliminated false-positive amplification from the wild-type allele. Further discrimination between true and false positives was enabled by using the quantification cycle (Cq) value of a single mutant template as a cut-off point, thus enabling robust distinction between true and false positives. As 10,000 JAK2 templates were used per replicate, the assay had a sensitivity of 1/10 -4 per replicate. Greater sensitivity could be reached by increasing the number of replicates analysed. Variation in replicates when low mutant-allele templates were present necessitated the use of a statistics-based approach to estimate the load of mutant JAK2 copies. QuanTAS-PCR showed comparable quantitative results when validated against a

Comparison of three human papillomavirus DNA detection methods: Next generation sequencing, multiplex-PCR and nested-PCR followed by Sanger based sequencing.

Science.gov (United States)

da Fonseca, Allex Jardim; Galvão, Renata Silva; Miranda, Angelica Espinosa; Ferreira, Luiz Carlos de Lima; Chen, Zigui

2016-05-01

To compare the diagnostic performance for HPV infection using three laboratorial techniques. Ninty-five cervicovaginal samples were randomly selected; each was tested for HPV DNA and genotypes using 3 methods in parallel: Multiplex-PCR, the Nested PCR followed by Sanger sequencing, and the Next_Gen Sequencing (NGS) with two assays (NGS-A1, NGS-A2). The study was approved by the Brazilian National IRB (CONEP protocol 16,800). The prevalence of HPV by the NGS assays was higher than that using the Multiplex-PCR (64.2% vs. 45.2%, respectively; P = 0.001) and the Nested-PCR (64.2% vs. 49.5%, respectively; P = 0.003). NGS also showed better performance in detecting high-risk HPV (HR-HPV) and HPV16. There was a weak interobservers agreement between the results of Multiplex-PCR and Nested-PCR in relation to NGS for the diagnosis of HPV infection, and a moderate correlation for HR-HPV detection. Both NGS assays showed a strong correlation for detection of HPVs (k = 0.86), HR-HPVs (k = 0.91), HPV16 (k = 0.92) and HPV18 (k = 0.91). NGS is more sensitive than the traditional Sanger sequencing and the Multiplex PCR to genotype HPVs, with promising ability to detect multiple infections, and may have the potential to establish an alternative method for the diagnosis and genotyping of HPV. © 2015 Wiley Periodicals, Inc.

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...

Diagnóstico temprano en un brote epidémico del virus Dengue en Piura usando RT-PCR y nested-PCR

Directory of Open Access Journals (Sweden)

Oscar Nolasco

1997-07-01

Full Text Available Un test de diagnóstico temprano (RT-PCR y Nested-PCR fue evaluado y comparado con métodos convencionales (cultivo in vitro, IFI y MAC-ELISA. Treinta y cuatro sueros de pacientes correspondientes de un brote epidémico de la costa norte peruana (Mancora, Piura en mayo de 1997 fueron incluidos en este estudio. Todos los sueros fueron obtenidos de pacientes que presentaron en los primeros cinco días manifestaciones clínicas siendo diagnosticados luego como dengue serotipo 1. Asimismo, RT-PCR permitió diagnosticar 82% de los sueros (28/34, sin embargo Mac-ELISA y cultivo in vitro reconocieron unicamente 41% de los sueros (14/34 y 38% de los sueros (13/34 respectivamente. Por lo tanto, el uso de esta herramienta molecular (RT-PCR y Nested-PCR permitiró dar un diagnóstico temprano a estos pacientes y actuar inmediatamente ante la presencia de un brote epidémico.

A vision and strategy for the virtual physiological human in 2010 and beyond

NARCIS (Netherlands)

Hunter, P.; Coveney, P.V.; de Bono, B.; Diaz, V.; Fenner, J.; Frangi, A.F.; Harris, P.; Hose, R.; Kohl, P.; Lawford, P.; McCormack, K.; Mendes, M.; Omholt, S.; Quarteroni, A.; Skar, J.; Tegner, J.; Thomas, S.R.; Tollis, I.; Tsamardinos, I.; van Beek, J.H.G.M.; Viceconti, M.

2010-01-01

European funding under framework 7 (FP7) for the virtual physiological human (VPH) project has been in place now for nearly 2 years. The VPH network of excellence (NoE) is helping in the development of common standards, open-source software, freely accessible data and model repositories, and various

A vision and strategy for the virtual physiological human: 2012 update

NARCIS (Netherlands)

Hunter, P.; Chapman, T.; Coveney, P.V.; De Bono, B.; Diaz, V.; Fenner, J.; Frangi, A.F.; Harris, P.; Hose, R.; Kohl, P.; Lawford, P.; McCormack, K.; Mendes, M.; Omholt, S.; Quarteroni, A.; Shublaq, N.; Skår, J.; Stroetmann, K.; Tegner, J.; Thomas, S.R.; Tollis, I.; Tsamardinos, I.; van Beek, J.H.G.M.; Viceconti, M.

2013-01-01

European funding under Framework 7 (FP7) for the virtual physiological human (VPH) project has been in place now for 5 years. The VPH Network of Excellence (NoE) has been set up to help develop common standards, open source software, freely accessible data and model repositories, and various

The Role of Model Fidelity in Model Predictive Control Based Hazard Avoidance in Unmanned Ground Vehicles Using Lidar Sensors

Science.gov (United States)

2013-03-08

34 VPH : A New Laser Radar Based Obstacle Avoidance Method for Intelligent Mobile Robots", Proceedings of World Congress on Intelligent Control and...Automation, Hangzhou, China, 5, pp. 4681- 4685. [9] Gong, J., Duan, Y., Man, Y., and Xiong, G., 2007, " VPH +: An Enhanced Vector Polar Histogram Method

Establishing a novel single-copy primer-internal intron-spanning PCR (spiPCR) procedure for the direct detection of gene doping.

Science.gov (United States)

Beiter, Thomas; Zimmermann, Martina; Fragasso, Annunziata; Armeanu, Sorin; Lauer, Ulrich M; Bitzer, Michael; Su, Hua; Young, William L; Niess, Andreas M; Simon, Perikles

2008-01-01

So far, the abuse of gene transfer technology in sport, so-called gene doping, is undetectable. However, recent studies in somatic gene therapy indicate that long-term presence of transgenic DNA (tDNA) following various gene transfer protocols can be found in DNA isolated from whole blood using conventional PCR protocols. Application of these protocols for the direct detection of gene doping would require almost complete knowledge about the sequence of the genetic information that has been transferred. Here, we develop and describe the novel single-copy primer-internal intron-spanning PCR (spiPCR) procedure that overcomes this difficulty. Apart from the interesting perspectives that this spiPCR procedure offers in the fight against gene doping, this technology could also be of interest in biodistribution and biosafety studies for gene therapeutic applications.

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.

Encapsulación de moléculas pequeñas mediante la precipitación salina de poliuretanos catioméricos

OpenAIRE

Fernández d’Arlas, Borja; Corcuera, María Ángeles; Eceiza, Arantxa

2015-01-01

En este trabajo se estudia un copolímero de poliuretano catiomérico (PU) con alta proporción de uretano como agente encapsulante de fármacos modelos (FM) mediante la encapsulación inducida por precipitación salina del PU y FM a pH < pI del PU. Mediante espectroscopia UV-Vis se ha estimado el porcentaje de encapsulación de varios FMs proponiéndose un modelo semi-empírico para determinar la distribución observada en las eficiencias de encapsulación, E, en función de su volumen molar...

Oncogenes E6-E7 de los Papilomavirus Humanos de alto riesgo detectados por PCR en Biopsias de pene incluidas en parafina

Directory of Open Access Journals (Sweden)

I Guerrero

1999-01-01

Full Text Available La alta prevalencia del papilomavirus humano (PVH, referida a nivel mundial, en lesiones genitales de ambos sexos, el rol del varón como reservorio pasivo del virus, y el incremento de la mortalidad por cáncer genital en la mujer en nuestro país, motiva la detección y correlación de los oncogenes de los PVH de alto riesgo con la neoplasia de pene. Informamos de diez casos de biopsias de carcinoma escamoso de pene, incluidos en parafina, los cuales fueron investigados para la presencia de los oncogenes E6-E7 de PVH de alto riesgo, utilizando cebadores tipo específico para los PVH -16 y 18, mediante la reacción en cadena de la polimerasa (PCR. El 40% de los casos mostró un producto de amplificación ADN E6-E7 de los PVH estudiados, correspondiendo el 75% de ellos a detección simple por PVH-18 y el 25% presentó detección mixta ADN E6 - E7 del PVH-16 y 18 simultáneamente. El producto de amplificación fue sometido a comprobación por análisis de restricción específico. La prevalencia obtenida de los oncogenes E6-E7 de los PVH de alto riesgo, usando un método tan sensible como la PCR, apoya el rol de estos virus en el proceso de carcinogénesis de la neoplasia de pene.

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...

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.

Papiloma bucal en pacientes Pediátricos: Potencial Transmisión Materna

OpenAIRE

Harris Ricardo, Jonathan; Rebolledo Cobos, Martha; Fortich Mesa, Natalia

2012-01-01

La evidencia actual afirma que el virus del papiloma humano (VPH) se puede transmitir tanto por vía sexual como por otras vías. Cuando el método de contagio es no sexual la madre parece ser el principal transmisor del VPH al recién nacido. Diversos autores en sus estudios reportan que se detectó ADN del VPH en el líquido amniótico, cordón umbilical, placenta y membranas fetales, lo que sugiere que la madre puede infectar al infante en la etapa de gestación o durante el parto. Después de la tr...

Transmitted wavefront error of a volume phase holographic grating at cryogenic temperature.

Science.gov (United States)

Lee, David; Taylor, Gordon D; Baillie, Thomas E C; Montgomery, David

2012-06-01

This paper describes the results of transmitted wavefront error (WFE) measurements on a volume phase holographic (VPH) grating operating at a temperature of 120 K. The VPH grating was mounted in a cryogenically compatible optical mount and tested in situ in a cryostat. The nominal root mean square (RMS) wavefront error at room temperature was 19 nm measured over a 50 mm diameter test aperture. The WFE remained at 18 nm RMS when the grating was cooled. This important result demonstrates that excellent WFE performance can be obtained with cooled VPH gratings, as required for use in future cryogenic infrared astronomical spectrometers planned for the European Extremely Large Telescope.

Rotational Isomers, Intramolecular Hydrogen Bond, and IR Spectra of o-Vinylphenol Homologs

Science.gov (United States)

Glazunov, V. P.; Berdyshev, D. V.; Balaneva, N. N.; Radchenko, O. S.; Novikov, V. L.

2018-03-01

The ν(OH) stretching-mode bands in solution IR spectra of five o-vinylphenol (o-VPh) homologs in the slightly polar solvents CCl4 and n-hexane were studied. Several rotamers with free OH groups were found in solutions of o-VPh and its methyl-substituted derivatives in n-hexane. The proportion of rotamers in o-VPh homologs with intramolecular hydrogen bonds (IHBs) O-H...π varied from 22 to 97% in the gas and cyclohexane according to B3LYP/cc-pVTZ calculations. The theoretically estimated effective enthalpies -ΔH of their IHBs varied in the range 0.20-2.24 kcal/mol.

Optimizing Noncovalent Interactions Between Lignin and Synthetic Polymers to Develop Effective Compatibilizers

Energy Technology Data Exchange (ETDEWEB)

Henry, Nathan [University of Tennessee, Knoxville (UTK); Harper, David [University of Tennessee, Knoxville (UTK), Center for Renewable Carbon; Dadmun, Mark D [ORNL

2012-01-01

Experiments are designed and completed to identify an effective polymeric compatibilizer for lignin polystyrene blends. Copolymers of styrene and vinylphenol are chosen as the structure of the compatibilizer as the VPh unit can readily form intermolecular hydrogen bonds with the lignin molecule. Electron microscopy, thermal analysis, and neutron refl ectivity results demonstrate that among these compatibilizers, a copolymer of styrene and VPh with 20% 30% VPh most readily forms intermolecular interactions with the lignin molecule and results in the most well-dispersed blends with lignin. This behavior is explained by invoking the competition of intra- and intermolecular hydrogen bonding and functional group accessibility in forming intermolecular interactions.

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.

Power Packaging of Spray-Cooled SiC Devices for High Temperature and High Voltage Operation: Final Report

Science.gov (United States)

2008-07-01

the desired switching frequencies. * I Three- r1aL phase dc-ac ac-dc Vph converter # 4 convertr converter 1 # 2 # 3 * I r --- -- -- - 4I6l kV ACSIDE...J:-----------------.HWn.XEMEL ------- J WDC_ SI CEI.................... ... .. .. .............. .......... J . Fig. 4.1 Block diagram of a PCM4. Vph

An admissions system to select veterinary medical students with an interest in food animals and veterinary public health.

Science.gov (United States)

Haarhuis, Jan C M; Muijtjens, Arno M M; Scherpbier, Albert J J A; van Beukelen, Peter

2009-01-01

Interest in the areas of food animals (FA) and veterinary public health (VPH) appears to be declining among prospective students of veterinary medicine. To address the expected shortage of veterinarians in these areas, the Utrecht Faculty of Veterinary Medicine has developed an admissions procedure to select undergraduates whose aptitude and interests are suited to these areas. A study using expert meetings, open interviews, and document analysis identified personal characteristics that distinguished veterinarians working in the areas of FA and VPH from their colleagues who specialized in companion animals (CA) and equine medicine (E). The outcomes were used to create a written selection tool. We validated this tool in a study among undergraduate veterinary students in their final (sixth) year before graduation. The applicability of the tool was verified in a study among first-year students who had opted to pursue either FA/VPH or CA/E. The tool revealed statistically significant differences with acceptable effect sizes between the two student groups. Because the written selection tool did not cover all of the differences between the veterinarians who specialized in FA/VPH and those who specialized in CA/E, we developed a prestructured panel interview and added it to the questionnaire. The evaluation of the written component showed that it was suitable for selecting those students who were most likely to succeed in the FA/VPH track.

Interlaboratory diagnostic accuracy of a Salmonella specific PCR-based method

DEFF Research Database (Denmark)

Malorny, B.; Hoorfar, Jeffrey; Hugas, M.

2003-01-01

A collaborative study involving four European laboratories was conducted to investigate the diagnostic accuracy of a Salmonella specific PCR-based method, which was evaluated within the European FOOD-PCR project (http://www.pcr.dk). Each laboratory analysed by the PCR a set of independent obtained...... presumably naturally contaminated samples and compared the results with the microbiological culture method. The PCR-based method comprised a preenrichment step in buffered peptone water followed by a thermal cell lysis using a closed tube resin-based method. Artificially contaminated minced beef and whole......-based diagnostic methods and is currently proposed as international standard document....

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...

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...

Human papillomavirus detection and typing using a nested-PCR-RFLP assay.

Science.gov (United States)

Coser, Janaina; Boeira, Thaís da Rocha; Fonseca, André Salvador Kazantzi; Ikuta, Nilo; Lunge, Vagner Ricardo

2011-01-01

It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner. Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene. Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay. The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5% increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay. The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

Science.gov (United States)

Espy, M. J.; Uhl, J. R.; Sloan, L. M.; Buckwalter, S. P.; Jones, M. F.; Vetter, E. A.; Yao, J. D. C.; Wengenack, N. L.; Rosenblatt, J. E.; Cockerill, F. R.; Smith, T. F.

2006-01-01

Real-time PCR has revolutionized the way clinical microbiology laboratories diagnose many human microbial infections. This testing method combines PCR chemistry with fluorescent probe detection of amplified product in the same reaction vessel. In general, both PCR and amplified product detection are completed in an hour or less, which is considerably faster than conventional PCR detection methods. Real-time PCR assays provide sensitivity and specificity equivalent to that of conventional PCR combined with Southern blot analysis, and since amplification and detection steps are performed in the same closed vessel, the risk of releasing amplified nucleic acids into the environment is negligible. The combination of excellent sensitivity and specificity, low contamination risk, and speed has made real-time PCR technology an appealing alternative to culture- or immunoassay-based testing methods for diagnosing many infectious diseases. This review focuses on the application of real-time PCR in the clinical microbiology laboratory. PMID:16418529

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...

Uncooled Split-off Quantum Infrared Sensors for 3-5 Micron Imaging Applications

Science.gov (United States)

2012-12-20

15 (b) (arrow), gradually raising the potential in Contact 1 to be higher than the emitter and Contact 2, until a built-in potential difference Vph ...presence of a graded barrier, a net flow of holes occurs, as indicated by the arrow, until a new equilibrium with a built-in potential Vph is

An in situ vapour phase hydrothermal surface doping approach for fabrication of high performance Co3O4 electrocatalysts with an exceptionally high S-doped active surface.

Science.gov (United States)

Tan, Zhijin; Liu, Porun; Zhang, Haimin; Wang, Yun; Al-Mamun, Mohammad; Yang, Hua Gui; Wang, Dan; Tang, Zhiyong; Zhao, Huijun

2015-04-04

A facile in situ vapour phase hydrothermal (VPH) surface doping approach has been developed for fabrication of high performance S-doped Co3O4 electrocatalysts with an unprecedentedly high surface S content (>47%). The demonstrated VPH doping approach could be useful for enrichment of surface active sites for other metal oxide electrocatalysts.

Study comparing human papillomavirus (HPV) real-time multiplex PCR and Hybrid Capture II INNO-LiPA v2 HPV genotyping PCR assays

DEFF Research Database (Denmark)

Iftner, Thomas; Germ, Liesje; Swoyer, Ryan

2009-01-01

methods has not been well characterized. Clinically, cytology is used to establish possible HPV infection. We evaluated the sensitivity and specificity of HPV multiplex PCR assays compared to those of the testing scheme of the Hybrid Capture II (HCII) assay followed by an HPV PCR/line hybridization assay...... (HCII-LiPA v2). SurePath residual samples were split into two aliquots. One aliquot was subjected to HCII testing followed by DNA extraction and LiPA v2 genotyping. The second aliquot was shipped to a second laboratory, where DNA was extracted and HPV multiplex PCR testing was performed. Comparisons...... were evaluated for 15 HPV types common in both assays. A slightly higher proportion of samples tested positive by the HPV multiplex PCR than by the HCII-LiPA v2 assay. The sensitivities of the multiplex PCR assay relative to those of the HCII-LiPA v2 assay for HPV types 6, 11, 16, and 18, for example...

A comparison of QuantStudio™ 3D Digital PCR and ARMS-PCR for measuring plasma EGFR T790M mutations of NSCLC patients

Directory of Open Access Journals (Sweden)

Feng Q

2018-01-01

Full Text Available Qin Feng,1,* Fei Gai,2,* Yaxiong Sang,2 Jie Zhang,3 Ping Wang,1 Yue Wang,1 Bing Liu,2 Dongmei Lin,1 Yang Yu,2 Jian Fang3 1Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education, Department of Pathology, Peking University Cancer Hospital & Institute, 2Oncology Business Division, Beijing Novogene Bioinformatics Technology Co., Ltd, 3Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education, Department of Thoracic Oncology II, Peking University Cancer Hospital & Institute, Beijing, China *These authors contributed equally to this work Background: 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.Purpose: 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.Patients and methods: 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.Results: 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 <0.1%. Limited by the amount of plasma DNA, 17 samples (AF <2.5% and eight samples (T790M- were selected for verification by ARMS-PCR. Four of those samples were verified by NGS as a third verification method. Among the selected 17 positive cases, ten samples presented mutant allele frequency <0.5%, and seven samples presented intermediate mutant allele frequency (0.5%＜AF＜2.5%. However, only three samples (3/17 were identified as positive by ARMS-PCR, namely, P6

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)

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.

High-throughput STR analysis for DNA database using direct PCR.

Science.gov (United States)

Sim, Jeong Eun; Park, Su Jeong; Lee, Han Chul; Kim, Se-Yong; Kim, Jong Yeol; Lee, Seung Hwan

2013-07-01

Since the Korean criminal DNA database was launched in 2010, we have focused on establishing an automated DNA database profiling system that analyzes short tandem repeat loci in a high-throughput and cost-effective manner. We established a DNA database profiling system without DNA purification using a direct PCR buffer system. The quality of direct PCR procedures was compared with that of conventional PCR system under their respective optimized conditions. The results revealed not only perfect concordance but also an excellent PCR success rate, good electropherogram quality, and an optimal intra/inter-loci peak height ratio. In particular, the proportion of DNA extraction required due to direct PCR failure could be minimized to <3%. In conclusion, the newly developed direct PCR system can be adopted for automated DNA database profiling systems to replace or supplement conventional PCR system in a time- and cost-saving manner. © 2013 American Academy of Forensic Sciences Published 2013. This article is a U.S. Government work and is in the public domain in the U.S.A.

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.

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…

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.

Analysis of the clonal relationship among clinical isolates of Salmonella enterica serovar Infantis by different typing methods Análisis de la relación clonal entre aislamientos clínicos de Salmonella enterica serovar Infantis mediante diferentes métodos de tipificación

Directory of Open Access Journals (Sweden)

Luis A. Merino

2003-06-01

Full Text Available Salmonella Infantis has been the second most common serovar in Argentina in the last two years, being isolated mostly from paediatric hospitalised patients. In order to determine the clonal relationship among Salmonella Infantis strains, we examined 15 isolates from paediatric patient faeces in Argentina (12 geographically related and 3 geographically non-related by using antimicrobial susceptibility, plasmid profiling, repetitive extragenic palindromic (REP PCR, enterobacterial repetitive intergenic consensus (ERIC PCR, and low-frequency restriction analysis of chromosomal DNA by pulsed field gel electrophoresis (PFGE. Four Spanish strains were included as controls of clonal diversity in molecular techniques. Antibiotype and plasmid profile was not useful as epidemiological tools. PFGE and REP-PCR were able to discriminate between Argentinean and Spanish isolates of Salmonella Infantis allowing to detect genetically related strains in three different cities. This finding indicates that a possible spread of a clone of this serovar in the North-eastern Region of Argentina has taken place in 1998.Salmonella Infantis ha sido el segundo serovar más común en la Argentina en los últimos dos años, siendo aislada principalmente, a partir de pacientes pediátricos hospitalizados. La relación clonal entre 15 aislamientos de Salmonella Infantis obtenidos de heces de pacientes pediátricos en Argentina se estudió mediante la susceptibilidad antimicrobiana, el perfil plasmídico, amplificación por reacción en cadena de la polimerasa (PCR de las secuencias repetitivas REP y ERIC, y electroforesis de ADN total en campo pulsátil (PFGE. Cuatro cepas españolas fueron incluidas como control de diversidad clonal. El antibiotipo y el perfil plasmídico no fueron herramientas útiles en la tipificación. PFGE y REP-PCR fueron capaces de discriminar entre las cepas argentinas y españolas de Salmonella Infantis, permitiendo detectar cepas gen

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

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.

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.

A comparative study of digital PCR and real-time qPCR for the detection and quantification of HPV mRNA in sentinel lymph nodes of cervical cancer patients.

Science.gov (United States)

Carow, Katrin; Read, Christina; Häfner, Norman; Runnebaum, Ingo B; Corner, Adam; Dürst, Matthias

2017-10-30

Qualitative analyses showed that the presence of HPV mRNA in sentinel lymph nodes of cervical cancer patients with pN0 status is associated with significantly decreased recurrence free survival. To further address the clinical potential of the strategy and to define prognostic threshold levels it is necessary to use a quantitative assay. Here, we compare two methods of quantification: digital PCR and standard quantitative PCR. Serial dilutions of 5 ng-5 pg RNA (≙ 500-0.5 cells) of the cervical cancer cell line SiHa were prepared in 5 µg RNA of the HPV-negative human keratinocyte cell line HaCaT. Clinical samples consisted of 10 sentinel lymph nodes with varying HPV transcript levels. Reverse transcription of total RNA (5 µg RNA each) was performed in 100 µl and cDNA aliquots were analyzed by qPCR and dPCR. Digital PCR was run in the RainDrop ® Digital PCR system (RainDance Technologies) using a probe-based detection of HPV E6/E7 cDNA PCR products with 11 µl template. qPCR was done using a Rotor Gene Q 5plex HRM (Qiagen) amplifying HPV E6/E7 cDNA in a SYBR Green format with 1 µl template. For the analysis of both, clinical samples and serial dilution samples, dPCR and qPCR showed comparable sensitivity. With regard to reproducibility, both methods differed considerably, especially for low template samples. Here, we found with qPCR a mean variation coefficient of 126% whereas dPCR enabled a significantly lower mean variation coefficient of 40% (p = 0.01). Generally, we saw with dPCR a substantial reduction of subsampling errors, which most likely reflects the large cDNA amounts available for analysis. Compared to real-time PCR, dPCR shows higher reliability. Thus, our HPV mRNA dPCR assay holds promise for the clinical evaluation of occult tumor cells in histologically tumor-free lymph nodes in future studies.

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

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...

Películas orgánicas delgadas preparadas mediante diversos métodos: propiedades ópticas, morfológicas y eléctricas

OpenAIRE

Pérez-Morales, M.

2005-01-01

En la presente Memoria se estudia la organización molecular de compuestos orgánicos, tales como derivados de porfirinas y C60, en películas de Langmuir formadas en la interfase aire-agua. Asimismo, se han depositado películas de derivados de porfirina sobre electrodos ITO mediante métodos electroquímicos. Por último, se han estudiado las propiedades fluorescentes de películas de porfirina preparadas mediante diversos métodos, para su posterior aplicación a la preparación de dispositivos elect...

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...

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 ...

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.

Percentile-based Weibull diameter distribution model for Pinus ...

African Journals Online (AJOL)

Using a site index equation and stem volume model developed for Pinus kesiya in the Philippines, a yield prediction system was created to predict the volume per ha (VPH) for each diameter class and, subsequently, the total volume of a stand. To evaluate the yield prediction system, the predicted mean VPH for each ...

Viability qPCR, a new tool for Legionella risk management.

Science.gov (United States)

Lizana, X; López, A; Benito, S; Agustí, G; Ríos, M; Piqué, N; Marqués, A M; Codony, F

2017-11-01

Viability quantitative Polymerase Chain Reaction (v-qPCR) is a recent analytical approach for only detecting live microorganisms by DNA amplification-based methods This approach is based on the use of a reagent that irreversibly fixes dead cells DNA. In this study, we evaluate the utility of v-qPCR versus culture method for Legionellosis risk management. The present study was performed using 116 real samples. Water samples were simultaneously analysed by culture, v-qPCR and qPCR methods. Results were compared by means of a non-parametric test. In 11.6% of samples using both methods (culture method and v-qPCR) results were positive, in 50.0% of samples both methods gave rise to negative results. As expected, equivalence between methods was not observed in all cases, as in 32.1% of samples positive results were obtained by v-qPCR and all of them gave rise to negative results by culture. Only in 6.3% of samples, with very low Legionella levels, was culture positive and v-qPCR negative. In 3.5% of samples, overgrowth of other bacteria did not allow performing the culture. When comparing both methods, significant differences between culture and v-qPCR were in the samples belonging to the cooling towers-evaporative condensers group. The v-qPCR method detected greater presence and obtained higher concentrations of Legionella spp. (p<0.001). Otherwise, no significant differences between methods were found in the rest of the groups. The v-qPCR method can be used as a quick tool to evaluate Legionellosis risk, especially in cooling towers-evaporative condensers, where this technique can detect higher levels than culture. The combined interpretation of PCR results along with the ratio of live cells is proposed as a tool for understanding the sample context and estimating the Legionellosis risk potential according to 4 levels of hierarchy. Copyright © 2017 Elsevier GmbH. All rights reserved.

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.

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

Electron Acoustic Waves in Pure Ion Plasmas

Science.gov (United States)

Anderegg, F.; Driscoll, C. F.; Dubin, D. H. E.; O'Neil, T. M.

2009-11-01

Electron Acoustic Waves (EAW) are the low frequency branch of electrostatic plasma waves. These waves exist in neutralized plasmas, pure electron plasmas and in pure ion plasmasfootnotetextF. Anderegg et al., PRL 102, 095001 (2009) and PoP 16, 055705 (2009). (where the name is deceptive). Here, we observe standing mθ= 0 mz= 1 EAWs in a pure ion plasma column. At small amplitude, the EAWs have a phase velocity vph ˜1.4 v, and the frequencies are in close agreement with theory. At moderate amplitudes, waves can be excited over a broad range of frequencies, with observed phase velocities in the range of 1.4 v vph diagnostic shows that particles slower than vph oscillate in phase with the wave, while particles moving faster than vph oscillate 180^o out of phase with the wave. From a fluid perspective, this gives an unusual negative dynamical compressibility. That is, the wave pressure oscillations are 180^o out of phase from the density oscillations, almost fully canceling the electrostatic restoring force, giving the low and malleable frequency.

Proton Linear Energy Transfer measurement using Emulsion Cloud Chamber

Energy Technology Data Exchange (ETDEWEB)

Shin, Jae-ik [Proton Therapy Center, National Cancer Center (Korea, Republic of); Division of Heavy Ion Clinical Research, Korea Institute of Radiological & Medical Sciences (KIRAMS), Seoul (Korea, Republic of); Park, Seyjoon [Department of Radiation Oncology, Samsung Medical Center, Sungkyunkwan University, School of Medicine, Seoul (Korea, Republic of); Kim, Haksoo; Kim, Meyoung [Proton Therapy Center, National Cancer Center (Korea, Republic of); Jeong, Chiyoung [Department of Radiation Oncology, Asan Medical Center, University of Ulsan College of Medicine, Seoul (Korea, Republic of); Cho, Sungkoo [Department of Radiation Oncology, Samsung Medical Center, Sungkyunkwan University, School of Medicine, Seoul (Korea, Republic of); Lim, Young Kyung; Shin, Dongho [Proton Therapy Center, National Cancer Center (Korea, Republic of); Lee, Se Byeong, E-mail: sblee@ncc.re.kr [Proton Therapy Center, National Cancer Center (Korea, Republic of); Morishima, Kunihiro; Naganawa, Naotaka; Sato, Osamu [Department of Physics, Nagoya University, Nagoya (Japan); Kwak, Jungwon [Department of Radiation Oncology, Asan Medical Center, University of Ulsan College of Medicine, Seoul (Korea, Republic of); Kim, Sung Hyun [Center for Underground Physics, Institute for Basic Science (IBS), Daejeon (Korea, Republic of); Cho, Jung Sook [Department of refinement education, Dongseo University, Busan (Korea, Republic of); Ahn, Jung Keun [Department of Physics, Korea University, Seoul (Korea, Republic of); Kim, Ji Hyun; Yoon, Chun Sil [Gyeongsang National University, Jinju (Korea, Republic of); Incerti, Sebastien [CNRS, IN2P3, CENBG, UMR 5797, F-33170 Gradignan (France); Université Bordeaux 1, CENBG, UMR 5797, F-33170 Gradignan (France)

2015-04-15

This study proposes to determine the correlation between the Volume Pulse Height (VPH) measured by nuclear emulsion and Linear Energy Transfer (LET) calculated by Monte Carlo simulation based on Geant4. The nuclear emulsion was irradiated at the National Cancer Center (NCC) with a therapeutic proton beam and was installed at 5.2 m distance from the beam nozzle structure with various thicknesses of water-equivalent material (PMMA) blocks to position with specific positions along the Bragg curve. After the beam exposure and development of the emulsion films, the films were scanned by S-UTS developed in Nagoya University. The proton tracks in the scanned films were reconstructed using the ‘NETSCAN’ method. Through this procedure, the VPH can be derived from each reconstructed proton track at each position along the Bragg curve. The VPH value indicates the magnitude of energy loss in proton track. By comparison with the simulation results obtained using Geant4, we found the correlation between the LET calculated by Monte Carlo simulation and the VPH measured by the nuclear emulsion.

Magnetic Reconnection Processes Involving Modes Propagating in the Ion Diamagnetic Velocity Direction

Science.gov (United States)

Buratti, P.; Coppi, B.; Pucella, G.; Zhou, T.

2013-10-01

Experiments in weakly collisional plasma regimes, (e.g. neutral beam heated plasmas in the H-regime), measuring the Doppler shift associated with the plasma local rotation, have shown that the toroidal mode phase velocity vph in the frame with Er = 0 is in the direction of the ion diamagnetic velocity. For ohmically heated plasmas, with higher collisionalities, vph in the laboratory frame is in the direction of the electron diamagnetic velocity, but plasma rotation is reversed as well, and vph, in the Er = 0 frame, is in the ion diamagnetic velocity direction. Theoretically, two classes of reconnecting modes should emerge: drift-tearing modes and ``inductive modes'' that depend on the effects of a finite plasma inductivity. The former modes, with vph in the direction of the electron diamagnetic velocity, require the pre-excitation of a different kind of mode in order to become unstable in weakly collisional regimes. The second kind of modes has a growth rate associated with the relevant finite ion viscosity. A comprehensive theory is presented. Sponsored in part by the US DOE.

Proton Linear Energy Transfer measurement using Emulsion Cloud Chamber

International Nuclear Information System (INIS)

Shin, Jae-ik; Park, Seyjoon; Kim, Haksoo; Kim, Meyoung; Jeong, Chiyoung; Cho, Sungkoo; Lim, Young Kyung; Shin, Dongho; Lee, Se Byeong; Morishima, Kunihiro; Naganawa, Naotaka; Sato, Osamu; Kwak, Jungwon; Kim, Sung Hyun; Cho, Jung Sook; Ahn, Jung Keun; Kim, Ji Hyun; Yoon, Chun Sil; Incerti, Sebastien

2015-01-01

This study proposes to determine the correlation between the Volume Pulse Height (VPH) measured by nuclear emulsion and Linear Energy Transfer (LET) calculated by Monte Carlo simulation based on Geant4. The nuclear emulsion was irradiated at the National Cancer Center (NCC) with a therapeutic proton beam and was installed at 5.2 m distance from the beam nozzle structure with various thicknesses of water-equivalent material (PMMA) blocks to position with specific positions along the Bragg curve. After the beam exposure and development of the emulsion films, the films were scanned by S-UTS developed in Nagoya University. The proton tracks in the scanned films were reconstructed using the ‘NETSCAN’ method. Through this procedure, the VPH can be derived from each reconstructed proton track at each position along the Bragg curve. The VPH value indicates the magnitude of energy loss in proton track. By comparison with the simulation results obtained using Geant4, we found the correlation between the LET calculated by Monte Carlo simulation and the VPH measured by the nuclear emulsion

Proton Linear Energy Transfer measurement using Emulsion Cloud Chamber

Science.gov (United States)

Shin, Jae-ik; Park, Seyjoon; Kim, Haksoo; Kim, Meyoung; Jeong, Chiyoung; Cho, Sungkoo; Lim, Young Kyung; Shin, Dongho; Lee, Se Byeong; Morishima, Kunihiro; Naganawa, Naotaka; Sato, Osamu; Kwak, Jungwon; Kim, Sung Hyun; Cho, Jung Sook; Ahn, Jung Keun; Kim, Ji Hyun; Yoon, Chun Sil; Incerti, Sebastien

2015-04-01

This study proposes to determine the correlation between the Volume Pulse Height (VPH) measured by nuclear emulsion and Linear Energy Transfer (LET) calculated by Monte Carlo simulation based on Geant4. The nuclear emulsion was irradiated at the National Cancer Center (NCC) with a therapeutic proton beam and was installed at 5.2 m distance from the beam nozzle structure with various thicknesses of water-equivalent material (PMMA) blocks to position with specific positions along the Bragg curve. After the beam exposure and development of the emulsion films, the films were scanned by S-UTS developed in Nagoya University. The proton tracks in the scanned films were reconstructed using the 'NETSCAN' method. Through this procedure, the VPH can be derived from each reconstructed proton track at each position along the Bragg curve. The VPH value indicates the magnitude of energy loss in proton track. By comparison with the simulation results obtained using Geant4, we found the correlation between the LET calculated by Monte Carlo simulation and the VPH measured by the nuclear emulsion.

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.

EL APRENDIZAJE DE LOS CONCEPTOS DE FUERZAS INTERMOLECULARES E INTRAMOLECULARES MEDIANTE LA MODELIZACIÓN DIDÁCTICA

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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.

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.

Applicazioni della PCR e PCR in situ nella diagnosi di infezioni batteriche e virali da biopsie fissate in formalina e incluse in paraffina

Directory of Open Access Journals (Sweden)

Stefania Cazzavillan

2003-03-01

Full Text Available In situ PCR, amplification of target DNA sequences in fixed cells, is a very useful molecular biology tecnique with potential to combine the high sensitivity of tube PCR with the precise anatomical localization of the targeted bsequences. It allows the study of low copy viral or bacterial DNA. In this study we document the utility of directin situ PCR with single primer pair by applying it to 3 infectious agents in different model systems: Borrelia burgdorferi in 5 Eritema migrans lesions and 55 primitive cutaneous B cell lymphomas, Chlamydia pneumoniae in 200 autoptic atheromasic lesions, and Papilloma virus in 20 CIN 1 (mild cervical dysplasia. In situ PCR seems to be a very promising tecnique; however, the prerequisite for the success of in situ PCR is conditioned by optimal standardization of the key variables which, on the other hand, are influenced by tissue composition.

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.

Real-Time PCR using a PCR Microchip with Integrated Thermal System and Polymer Waveguides for the Detection of Campylobacter jejuni

DEFF Research Database (Denmark)

Wang, Zhenyu; Sekulovic, Andrea; Kutter, Jörg Peter

2006-01-01

A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. By using the integrated optical system of the real-time PCR chip, cadF – a virulence gene of Campylobacter jejuni, could specifically be detected. Two different DNA binding dyes, SYTOX...

A compact multichannel spectrometer for Thomson scatteringa)

Science.gov (United States)

Schoenbeck, N. L.; Schlossberg, D. J.; Dowd, A. S.; Fonck, R. J.; Winz, G. R.

2012-10-01

The availability of high-efficiency volume phase holographic (VPH) gratings and intensified CCD (ICCD) cameras have motivated a simplified, compact spectrometer for Thomson scattering detection. Measurements of Te VPH grating and measurements Te > 100 eV by a 2072 l/mm VPH grating. The spectrometer uses a fast-gated (˜2 ns) ICCD camera for detection. A Gen III image intensifier provides ˜45% quantum efficiency in the visible region. The total read noise of the image is reduced by on-chip binning of the CCD to match the 8 spatial channels and the 10 spectral bins on the camera. Three spectrometers provide a minimum of 12 spatial channels and 12 channels for background subtraction.

A compact multichannel spectrometer for Thomson scattering.

Science.gov (United States)

Schoenbeck, N L; Schlossberg, D J; Dowd, A S; Fonck, R J; Winz, G R

2012-10-01

The availability of high-efficiency volume phase holographic (VPH) gratings and intensified CCD (ICCD) cameras have motivated a simplified, compact spectrometer for Thomson scattering detection. Measurements of T(e) VPH grating and measurements T(e) > 100 eV by a 2072 l∕mm VPH grating. The spectrometer uses a fast-gated (~2 ns) ICCD camera for detection. A Gen III image intensifier provides ~45% quantum efficiency in the visible region. The total read noise of the image is reduced by on-chip binning of the CCD to match the 8 spatial channels and the 10 spectral bins on the camera. Three spectrometers provide a minimum of 12 spatial channels and 12 channels for background subtraction.

Aplicación de técnicas de electrodeposición mediante pulsos de corriente para la obtención de recubrimientos metálicos

OpenAIRE

Imaz Molina, Naroa

2013-01-01

En la presente tesis doctoral se han aplicado herramientas quimiométricas en el estudio y optimización de los parámetros implicados en la electrodeposición de metales y aleaciones mediante pulsos de corriente, centrando el trabajo en dos procesos determinados: • Cromo duro: con objeto de mejorar la funcionalidad y durabilidad de estos recubrimientos tan extendidos industrialmente, se ha investigado el efecto de la electrodeposición mediante pulsos de corriente...

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 ...

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

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.

Development and validation of a quantitative PCR assay for Ichthyophonus spp.

Science.gov (United States)

White, Vanessa C; Morado, J Frank; Crosson, Lisa M; Vadopalas, Brent; Friedman, Carolyn S

2013-04-29

Members of the genus Ichthyophonus are trophically transmitted, cosmopolitan parasites that affect numerous fish species worldwide. A quantitative PCR (qPCR) assay specific for genus Ichthyophonus 18S ribosomal DNA was developed for parasite detection and surveillance. The new assay was tested for precision, repeatability, reproducibility, and both analytical sensitivity and specificity. Diagnostic sensitivity and specificity were estimated using tissue samples from a wild population of walleye pollock Theragra chalcogramma. Ichthyophonus sp. presence in tissue samples was determined by qPCR, conventional PCR (cPCR), and histology. Parasite prevalence estimates varied depending upon the detection method employed and tissue type tested. qPCR identified the greatest number of Ichthyophonus sp.-positive cases when applied to walleye pollock skeletal muscle. The qPCR assay proved sensitive and specific for Ichthyophonus spp. DNA, but like cPCR, is only a proxy for infection. When compared to cPCR, qPCR possesses added benefits of parasite DNA quantification and a 100-fold increase in analytical sensitivity. Because this novel assay is specific for known members of the genus, it is likely appropriate for detecting Ichthyophonus spp. DNA in various hosts from multiple regions. However, species-level identification and isotype variability would require DNA sequencing. In addition to distribution and prevalence applications, this assay could be modified and adapted for use with zooplankton or environmental samples. Such applications could aid in investigating alternate routes of transmission and life history strategies typical to members of the genus Ichthyophonus.

Technical aspects and recommendations for single-cell qPCR.

Science.gov (United States)

Ståhlberg, Anders; Kubista, Mikael

2018-02-01

Single cells are basic physiological and biological units that can function individually as well as in groups in tissues and organs. It is central to identify, characterize and profile single cells at molecular level to be able to distinguish different kinds, to understand their functions and determine how they interact with each other. During the last decade several technologies for single-cell profiling have been developed and used in various applications, revealing many novel findings. Quantitative PCR (qPCR) is one of the most developed methods for single-cell profiling that can be used to interrogate several analytes, including DNA, RNA and protein. Single-cell qPCR has the potential to become routine methodology but the technique is still challenging, as it involves several experimental steps and few molecules are handled. Here, we discuss technical aspects and provide recommendation for single-cell qPCR analysis. The workflow includes experimental design, sample preparation, single-cell collection, direct lysis, reverse transcription, preamplification, qPCR and data analysis. Detailed reporting and sharing of experimental details and data will promote further development and make validation studies possible. Efforts aiming to standardize single-cell qPCR open up means to move single-cell analysis from specialized research settings to standard research laboratories. Copyright © 2017 Elsevier Ltd. All rights reserved.

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.

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.

La creación de nuevas reglas técnicas en el IGBM mediante la Programación Genética

Directory of Open Access Journals (Sweden)

Fernando Fernández Rodríguez

2002-01-01

Full Text Available En este trabajo analizamos la capacidad de generar beneficios de reglas técnicas creadas mediante la programación genética en el Índice General de la Bolsa de Madrid . Esta nueva técnica, que no es mas que una expansión de los algoritmos genéticos, permite generar nuevas reglas de contratación en bolsa mediante procedimientos de optimización basados en la selección natural darwiniana. Se comparará los rendimientos obtenidos con la sencilla estrategia de comprar y mantener así como con las reglas basadas en medias móviles más comúnmente usadas en los mercados.

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.

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

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

Cytogenetic and molecular characteristics of 25 Chilean patients with a variant Ph translocation.

Science.gov (United States)

Legues, Maria E; Encina, Andrea; Valenzuela, Mercedes; Palma, Tamara; Undurraga, Maria S

2011-07-01

Chronic myeloid leukemia (CML) is characterized by the presence of the Philadelphia chromosome (Ph), which results from a balanced translocation between chromosomes 9 and 22, the t(9;22)(q34;q11.2). In 5-10% of the cases, variants of the Ph (vPh) are detected, involving various breakpoints in addition to 9q34 and 22q11.2. Deletions on the der(9) and der(22) can be detected in approximately 10-15% of CML patients. The frequency of a deletion of the der(9) in vPh CML is variable. Most studies have shown high frequencies (30-45%) in this subgroup. We report the cytogenetic evaluation of 25 vPh cases, which represents 6.8% of the CML cases diagnosed at one institution in 20 years. The breakpoints of the partners of the vPh in our patients agree with those reported previously, except for a novel 18q23. We found a low incidence of deletions of the der(9) (10%) and der(22) (5%) in these patients, contrasting with several reports in the literature. This finding may reflect the extensive spectrum of aberrations in vPh, and the possibility that a considerable group of these aberrations may not affect the genetic stability of 5'ABL1 and 3'BCR. Epidemiologic differences may also exist and could explain our results. These differences would require further investigation. Copyright © 2011 Elsevier Inc. All rights reserved.

Incidence of pulmonary aspergillosis and correlation of conventional diagnostic methods with nested PCR and real-time PCR assay using BAL fluid in intensive care unit patients.

Science.gov (United States)

Zarrinfar, Hossein; Makimura, Koichi; Satoh, Kazuo; Khodadadi, Hossein; Mirhendi, Hossein

2013-05-01

Although the incidence of invasive aspergillosis in the intensive care unit (ICU) is scarce, it has emerged as major problems in critically ill patients. In this study, the incidence of pulmonary aspergillosis (PA) in ICU patients has evaluated and direct microscopy and culture has compared with nested polymerase chain reaction (PCR) and real-time PCR for detection of Aspergillus fumigatus and A. flavus in bronchoalveolar lavage (BAL) samples of the patients. Thirty BAL samples obtained from ICU patients during a 16-month period were subjected to direct examinations on 20% potassium hydroxide (KOH) and culture on two culture media. Nested PCR targeting internal transcribed spacer ribosomal DNA and TaqMan real-time PCR assay targeting β-tubulin gene were used for the detection of A. fumigatus and A. flavus. Of 30 patients, 60% were men and 40% were women. The diagnosis of invasive PA was probable in 1 (3%), possible in 11 (37%), and not IPA in 18 (60%). Nine samples were positive in nested PCR including seven samples by A. flavus and two by A. fumigatus specific primers. The lowest amount of DNA that TaqMan real-time PCR could detect was ≥40 copy numbers. Only one of the samples had a positive result of A. flavus real-time PCR with Ct value of 37.5. Although a significant number of specimens were positive in nested PCR, results of this study showed that establishment of a correlation between the conventional methods with nested PCR and real-time PCR needs more data confirmed by a prospective study with a larger sample group. © 2013 Wiley Periodicals, Inc.

Interlaboratory study of DNA extraction from multiple ground samples, multiplex real-time PCR, and multiplex qualitative PCR for individual kernel detection system of genetically modified maize.

Science.gov (United States)

Akiyama, Hiroshi; Sakata, Kozue; Makiyma, Daiki; Nakamura, Kosuke; Teshima, Reiko; Nakashima, Akie; Ogawa, Asako; Yamagishi, Toru; Futo, Satoshi; Oguchi, Taichi; Mano, Junichi; Kitta, Kazumi

2011-01-01

In many countries, the labeling of grains, feed, and foodstuff is mandatory if the genetically modified (GM) organism content exceeds a certain level of approved GM varieties. We previously developed an individual kernel detection system consisting of grinding individual kernels, DNA extraction from the individually ground kernels, GM detection using multiplex real-time PCR, and GM event detection using multiplex qualitative PCR to analyze the precise commingling level and varieties of GM maize in real sample grains. We performed the interlaboratory study of the DNA extraction with multiple ground samples, multiplex real-time PCR detection, and multiplex qualitative PCR detection to evaluate its applicability, practicality, and ruggedness for the individual kernel detection system of GM maize. DNA extraction with multiple ground samples, multiplex real-time PCR, and multiplex qualitative PCR were evaluated by five laboratories in Japan, and all results from these laboratories were consistent with the expected results in terms of the commingling level and event analysis. Thus, the DNA extraction with multiple ground samples, multiplex real-time PCR, and multiplex qualitative PCR for the individual kernel detection system is applicable and practicable in a laboratory to regulate the commingling level of GM maize grain for GM samples, including stacked GM maize.

Modified DNA extraction for rapid PCR detection of methicillin-resistant staphylococci

International Nuclear Information System (INIS)

Japoni, A.; Alborzi, A.; Rasouli, M.; Pourabbas, B.

2004-01-01

Nosocomial infection caused by methicillin-resistant staphylococci poses a serious problem in many countries. The aim of this study was to rapidly and reliably detect methicillin-resistant-staphylococci in order to suggest appropriate therapy. The presence or absence of the methicillin-resistance gene in 115 clinical isolates of staphylococcus aureus and 50 isolates of coagulase negative staphylococci was examined by normal PCR. DNA extraction for PCR performance was then modified by omission of achromopeptadiase and proteinase K digestion, phenol/chloroform extraction and ethanol precipitation. All isolates with Mic>8 μ g/ml showed positive PCR. No differences in PCR detection have been observed when normal and modified DNA extractions have been performed. Our modified DNA extraction can quickly detect methicillin-resistant staphylococci by PCR. The advantage of rapid DNA extraction extends to both reduction of time and cost of PCR performance. This modified DNA extraction is suitable for different PCR detection, when staphylococci are the subject of DNA analysis

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.

Desarrollo de un escáner 3D mediante cámaras estereoscópicas e iluminación láser

OpenAIRE

Cristina, Federico; Dapoto, Sebastián H.; Vegas, Javier; Artola, Verónica; Russo, Claudia Cecilia; Abásolo Guerrero, María José

2007-01-01

Los dispositivos de escaneo tridimensional permiten obtener modelos de objetos utilizando distintas técnicas de captura. Esta tarea puede ser llevada a cabo por ejemplo mediante estereovisión, el cual es un método de reconstrucción 3D a partir de fotografías. Las técnicas de reconstrucción 3D mediante luz se basan en la proyección de un patrón de luz conocido sobre una escena y a partir del análisis de la proyección puede deducirse la forma de los objetos. De esta manera, basándose en la info...

An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection.

Science.gov (United States)

Pitz, Amanda de Faveri; Koishi, Andrea Cristine; Tavares, Eliandro Reis; Andrade, Fábio Goulart de; Loth, Eduardo Alexandre; Gandra, Rinaldo Ferreira; Venancio, Emerson José

2013-01-01

Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays.

Análisis de la biodiversidad de macroinvertebrados bentónicos del Río Cunas mediante indicadores ambientales, Junín - Perú

OpenAIRE

Custodio Villanueva, María

2013-01-01

El objetivo de la investigación fue analizar el estado de la biodiversidad de macroinvertebrados bentónicos del río Cunas mediante indicadores ambientales. Se utilizaron los métodos de observación, descripción y explicación; el tipo de investigación es básica y el diseño no experimental, de tipo longitudinal. Se definieron tres sectores de muestreo, San Blas, Huarisca y La Perla. La valoración de las presiones antrópicas se realizó mediante la determinación de DBO5 aportada por aguas residual...

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...

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 ...

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.

A MIQE-compliant real-time PCR assay for Aspergillus detection.

Directory of Open Access Journals (Sweden)

Gemma L Johnson

Full Text Available The polymerase chain reaction (PCR is widely used as a diagnostic tool in clinical laboratories and is particularly effective for detecting and identifying infectious agents for which routine culture and microscopy methods are inadequate. Invasive fungal disease (IFD is a major cause of morbidity and mortality in immunosuppressed patients, and optimal diagnostic criteria are contentious. Although PCR-based methods have long been used for the diagnosis of invasive aspergillosis (IA, variable performance in clinical practice has limited their value. This shortcoming is a consequence of differing sample selection, collection and preparation protocols coupled with a lack of standardisation of the PCR itself. Furthermore, it has become clear that the performance of PCR-based assays in general is compromised by the inadequacy of experimental controls, insufficient optimisation of assay performance as well as lack of transparency in reporting experimental details. The recently published "Minimum Information for the publication of real-time Quantitative PCR Experiments" (MIQE guidelines provide a blueprint for good PCR assay design and unambiguous reporting of experimental detail and results. We report the first real-time quantitative PCR (qPCR assay targeting Aspergillus species that has been designed, optimised and validated in strict compliance with the MIQE guidelines. The hydrolysis probe-based assay, designed to target the 18S rRNA DNA sequence of Aspergillus species, has an efficiency of 100% (range 95-107%, a dynamic range of at least six orders of magnitude and limits of quantification and detection of 6 and 0.6 Aspergillus fumigatus genomes, respectively. It does not amplify Candida, Scedosporium, Fusarium or Rhizopus species and its clinical sensitivity is demonstrated in histological material from proven IA cases, as well as concordant PCR and galactomannan data in matched broncho-alveolar lavage and blood samples. The robustness

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.

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)

Simultaneous detection of Legionella species and Legionella pneumophila by duplex PCR (dPCR assay in cooling tower water samples from Jakarta, Indonesia

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Andi Yasmon

2010-11-01

Full Text Available Aim: Since culture method is time-consuming and has low  sensitivity, we developed a duplex PCR (dPCR assay for the detection of Legionella sp. and L. pneumophila in cooling tower samples. We used culture method as a gold standard.Methods: Optimization of dPCR method was performed to obtain an assay with high sensitivity and specifi city. The optimized method was used to detect Legionella sp. dan L. pneumophila in 9 samples obtained from 9 buildings in Jakarta. For culture method, the bacteria were grown or isolated on selective growth factor supplemented-buffered charcoal yeast extract (BCYE media.Results: Of 9 samples tested by dPCR assay, 6 were positive for Legionella species,1 was positive for L. pneumophila, and 2 showed negative results. For the same samples, no Legionella sp. was detected by the culture method.Conclusion: dPCR assay was much more sensitive than the culture method and was potentially used as a rapid, specifi c and sensitive test for routine detection of Legionella sp. dan for L. pneumophila in water samples. (Med J Indones 2010; 19:223-7Keywords: BCYE media, mip gene, 16S-rRNA gene

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...

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 ...

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...

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...

An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

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Amanda de Faveri Pitz

2013-12-01

Full Text Available Introduction Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR assay for the detection of Paracoccidioides brasiliensis. Methods We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. Results The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. Conclusions The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays.

Análisis de la utilidad de la olfatogustometría mediante BAST-24 en la población diabética y su relación con la función renal

OpenAIRE

Gascón Rubio, María Cristina

2014-01-01

157 p. : il., graf. Realizamos un estudio descriptivo observacional del sentido del olfato mediante el test BAST-24 de la población diabética. Valoramos su función renal mediante determinaciones analíticas.

Detection of Fusarium verticillioides by PCR-ELISA based on FUM21 gene.

Science.gov (United States)

Omori, Aline Myuki; Ono, Elisabete Yurie Sataque; Bordini, Jaqueline Gozzi; Hirozawa, Melissa Tiemi; Fungaro, Maria Helena Pelegrinelli; Ono, Mario Augusto

2018-08-01

Fusarium verticillioides is a primary corn pathogen and fumonisin producer which is associated with toxic effects in humans and animals. The traditional methods for detection of fungal contamination based on morphological characteristics are time-consuming and show low sensitivity and specificity. Therefore, the objective of this study was to develop a PCR-ELISA based on the FUM21 gene for F. verticillioides detection. The DNA of the F. verticillioides, Fusarium sp., Aspergillus sp. and Penicillium sp. isolates was analyzed by conventional PCR and PCR-ELISA to determine the specificity. The PCR-ELISA was specific to F. verticillioides isolates, showed a 2.5 pg detection limit and was 100-fold more sensitive than conventional PCR. In corn samples inoculated with F. verticillioides conidia, the detection limit of the PCR-ELISA was 1 × 10 4 conidia/g and was also 100-fold more sensitive than conventional PCR. Naturally contaminated corn samples were analyzed by PCR-ELISA based on the FUM21 gene and PCR-ELISA absorbance values correlated positively (p PCR-ELISA developed in this study can be useful for F. verticillioides detection in corn samples. Copyright © 2018 Elsevier Ltd. All rights reserved.

Aspectos por considerar para una efectiva integración universitaria mediante las nuevas tecnologías. Una perspectiva desde la sociología de las organizaciones

OpenAIRE

Cordero, Aleska

2008-01-01

En el presente artículo se desarrollan algunas ideas y consideraciones por tomar en cuenta, desde el enfoque organizacional, para fundamentar una efectiva integración iberoamericana entre las instituciones universitarias mediante el uso de las nuevas tecnologías. Se presentan ciertos conceptos provenientes de la teoría organizacional (cultura organizacional, cultura profesional, resistencia al cambio) mediante los cuales es posible abordar el análisis de los problemas que se plantean en las i...

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

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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.

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

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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.

PCR IN TRAUMATOLOGY AND ORTHOPAEDICS: METHOD DESCRIPTION AND APPLICABILITY

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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.

The male role in cervical cancer El papel del varón en el cáncer cervical

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Xavier Castellsagué

2003-01-01

Full Text Available Experimental, clinical, and epidemiological evidence strongly suggests that genital Human Papillomaviruses (HPVs are predominantly sexually transmitted. Epidemiological studies in virginal and HPV-negative women clearly indicate that sexual intercourse is virtually a necessary step for acquiring HPV. As with any other sexually transmitted disease (STD men are implicated in the epidemiological chain of the infection. Penile HPVs are predominantly acquired through sexual contacts. Sexual contacts with women who are prostitutes play an important role in HPV transmission and in some populations sex workers may become an important reservoir of high-risk HPVs. Acting both as "carriers" and "vectors" of oncogenic HPVs male partners may markedly contribute to the risk of developing cervical cancer in their female partners. Thus, in the absence of screening programs, a woman's risk of cervical cancer may depend less on her own sexual behavior than on that of her husband or other male partners. Although more rarely than women, men may also become the "victims" of their own HPV infections as a fraction of infected men are at an increased risk of developing penile and anal cancers. Male circumcision status has been shown to reduce the risk not only of acquiring and transmitting genital HPVs but also of cervical cancer in their female partners. More research is needed to better understand the natural history and epidemiology of HPV infections in men.Evidencia experimental, clínica y epidemiológica demuestra que los papilomavirus humanos (VPH genitales son predominantemente de transmisión sexual. Estudios experimentales en mujeres vírgenes y en mujeres VPH-negativas indican de forma clara que el coito es virtualmente un paso necesario para adquirir el VPH. Como ocurre con cualquier otra infección de transmisión sexual (ITS los varones están implicados en la cadena epidemiológica de la infección. Los VPH en el pene son predominantemente adquiridos a

Clinical Assessment of a Nocardia PCR-Based Assay for Diagnosis of Nocardiosis.

Science.gov (United States)

Rouzaud, Claire; Rodriguez-Nava, Véronica; Catherinot, Emilie; Méchaï, Frédéric; Bergeron, Emmanuelle; Farfour, Eric; Scemla, Anne; Poirée, Sylvain; Delavaud, Christophe; Mathieu, Daniel; Durupt, Stéphane; Larosa, Fabrice; Lengelé, Jean-Philippe; Christophe, Jean-Louis; Suarez, Felipe; Lortholary, Olivier; Lebeaux, David

2018-06-01

The diagnosis of nocardiosis, a severe opportunistic infection, is challenging. We assessed the specificity and sensitivity of a 16S rRNA Nocardia PCR-based assay performed on clinical samples. In this multicenter study (January 2014 to April 2015), patients who were admitted to three hospitals and had an underlying condition favoring nocardiosis, clinical and radiological signs consistent with nocardiosis, and a Nocardia PCR assay result for a clinical sample were included. Patients were classified as negative control (NC) (negative Nocardia culture results and proven alternative diagnosis or improvement at 6 months without anti- Nocardia treatment), positive control (PC) (positive Nocardia culture results), or probable nocardiosis (positive Nocardia PCR results, negative Nocardia culture results, and no alternative diagnosis). Sixty-eight patients were included; 47 were classified as NC, 8 as PC, and 13 as probable nocardiosis. PCR results were negative for 35/47 NC patients (74%). For the 12 NC patients with positive PCR results, the PCR assay had been performed with respiratory samples. These NC patients had chronic bronchopulmonary disease more frequently than did the NC patients with negative PCR results (8/12 patients [67%] versus 11/35 patients [31%]; P = 0.044). PCR results were positive for 7/8 PC patients (88%). There were 13 cases of probable nocardiosis, diagnosed solely using the PCR results; 9 of those patients (69%) had lung involvement (consolidation or nodule). Nocardia PCR testing had a specificity of 74% and a sensitivity of 88% for the diagnosis of nocardiosis. Nocardia PCR testing may be helpful for the diagnosis of nocardiosis in immunocompromised patients but interpretation of PCR results from respiratory samples is difficult, because the PCR assay may also detect colonization. Copyright © 2018 American Society for Microbiology.

Environmental Assessment for US 98 (SR 30) at the Entrance to Hurlburt Field

Science.gov (United States)

2010-09-01

ppm) VPH Proposed Action 2012 30/50 3,650 Hurlburt Field main gate 10.0 35 6.0 9.0 Proposed Action 2032 30/50 4,830 Hurlburt Field main...NAAQS & FAAQS 8-Hr (ppm) VPH No Build 2012 30/45 4,770 Hurlburt Field main gate 16.1 35 9.7 9 No Build 2032 30/45 5,460 Hurlburt Field main

RT-PCR Detection of HIV in Republic of Macedonia

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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

Sources of PCR-induced distortions in high-throughput sequencing data sets

Science.gov (United States)

Kebschull, Justus M.; Zador, Anthony M.

2015-01-01

PCR permits the exponential and sequence-specific amplification of DNA, even from minute starting quantities. PCR is a fundamental step in preparing DNA samples for high-throughput sequencing. However, there are errors associated with PCR-mediated amplification. Here we examine the effects of four important sources of error—bias, stochasticity, template switches and polymerase errors—on sequence representation in low-input next-generation sequencing libraries. We designed a pool of diverse PCR amplicons with a defined structure, and then used Illumina sequencing to search for signatures of each process. We further developed quantitative models for each process, and compared predictions of these models to our experimental data. We find that PCR stochasticity is the major force skewing sequence representation after amplification of a pool of unique DNA amplicons. Polymerase errors become very common in later cycles of PCR but have little impact on the overall sequence distribution as they are confined to small copy numbers. PCR template switches are rare and confined to low copy numbers. Our results provide a theoretical basis for removing distortions from high-throughput sequencing data. In addition, our findings on PCR stochasticity will have particular relevance to quantification of results from single cell sequencing, in which sequences are represented by only one or a few molecules. PMID:26187991

Las redes SAPUVET y SPVet: un modelo de integración en materia de salud pública veterinaria entre Europa y América Latina The SAPUVET and SPVet networks: an integration model in veterinary public health between Europe and Latin America

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Carmelo Ortega

2005-01-01

Full Text Available This paper underscores the need for animal health professionals to play a more significant role in the sphere of public health, particularly during natural disasters and other emergency situations which can reduce the availability of safe foods from animal sources. In order to help readers understand the importance of the emerging field of veterinary public health (VPH, the authors review the importance and current status of VPH in different countries and assess the role that veterinarians can play in overcoming situations that threaten human health. The last section discusses the need for training veterinarians in VPH and the important role that veterinarians can play within international public health organizations and multidisciplinary groups, such as SAPUVET and SPVet networks.

Vapor-phase hydrothermal transformation of HTiOF3 intermediates into {001} faceted anatase single-crystalline nanosheets.

Science.gov (United States)

Liu, Porun; Wang, Yun; Zhang, Haimin; An, Taicheng; Yang, Huagui; Tang, Zhiyong; Cai, Weiping; Zhao, Huijun

2012-12-07

For the first time, a facile, one-pot hydrofluoric acid vapor-phase hydrothermal (HF-VPH) method is demonstrated to directly grow single-crystalline anatase TiO(2) nanosheets with 98.2% of exposed {001} faceted surfaces on the Ti substrate via a distinctive two-stage formation mechanism. The first stage produces a new intermediate crystal (orthorhombic HTiOF(3) ) that is transformed into anatase TiO(2) nanosheets during the second stage. The findings reveal that the HF-VPH reaction environment is unique and differs remarkably from that of liquid-phase hydrothermal processes. The uniqueness of the HF-VPH conditions can be readily used to effectively control the nanostructure growth. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

[A Duplex PCR Method for Detection of Babesia caballi and Theileria equi].

Science.gov (United States)

Zhang, Yang; Zhang, Yu-ting; Wang, Zhen-bao; Bolati; Li, Hai; Bayinchahan

2015-04-01

To develop a duplex PCR assay for detection of Babesia caballi and Theileria equi. Two pairs of primers were designed according to the BC48 gene of B. caballi and 18 s rRNA gene of T. equi, and a duplex PCR assay was developed by the optimization of reaction conditions. The specificity, sensitivity and reliability of the method were tested. The horse blood samples of suspected cases were collected from Yili region, and detected by the duplex PCR, microspopy, conventional PCR, and fluorescence quantitative PCR, and the results were compared. Using the duplex PCR assay, the specific fragments of 155 bp and 280 bp were amplified from DNA samples of B. caballi and T. equi, respectively. No specific fragment was amplified from DNA samples of B. bigemina, Theilerdia annulata, Theilerdia sergenti, Toxoplasma gondii, Neospora caninum, and Trypanosoma evansi. The limit of detection was 4.85 x 10(5) copies/L for B. caballi DNA and 4.85 x 10(4) copies/µl for T. equi DNA, respectively. Among the 24 blood samples, 11 were found B. caballi-positive by the duplex PCR assay, and 18 were T. equi-positive. The coincidence rate of microscopy, conventional PCR, and fluorescence quantitative PCR with duplex PCR was 91.7% (22/24), 95.8% (23/24), and 95.8% (23/24), respectively. A duplex PCR assay for simultaneous detection of B. caballi and T. equi is established.

Cross-platform comparison of SYBR® Green real-time PCR with TaqMan PCR, microarrays and other gene expression measurement technologies evaluated in the MicroArray Quality Control (MAQC study

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Dial Stacey L

2008-07-01

Full Text Available Abstract Background The MicroArray Quality Control (MAQC project evaluated the inter- and intra-platform reproducibility of seven microarray platforms and three quantitative gene expression assays in profiling the expression of two commercially available Reference RNA samples (Nat Biotechnol 24:1115-22, 2006. The tested microarrays were the platforms from Affymetrix, Agilent Technologies, Applied Biosystems, GE Healthcare, Illumina, Eppendorf and the National Cancer Institute, and quantitative gene expression assays included TaqMan® Gene Expression PCR Assay, Standardized (Sta RT-PCR™ and QuantiGene®. The data showed great consistency in gene expression measurements across different microarray platforms, different technologies and test sites. However, SYBR® Green real-time PCR, another common technique utilized by half of all real-time PCR users for gene expression measurement, was not addressed in the MAQC study. In the present study, we compared the performance of SYBR Green PCR with TaqMan PCR, microarrays and other quantitative technologies using the same two Reference RNA samples as the MAQC project. We assessed SYBR Green real-time PCR using commercially available RT2 Profiler™ PCR Arrays from SuperArray, containing primer pairs that have been experimentally validated to ensure gene-specificity and high amplification efficiency. Results The SYBR Green PCR Arrays exhibit good reproducibility among different users, PCR instruments and test sites. In addition, the SYBR Green PCR Arrays have the highest concordance with TaqMan PCR, and a high level of concordance with other quantitative methods and microarrays that were evaluated in this study in terms of fold-change correlation and overlap of lists of differentially expressed genes. Conclusion These data demonstrate that SYBR Green real-time PCR delivers highly comparable results in gene expression measurement with TaqMan PCR and other high-density microarrays.

A quantitative TaqMan PCR assay for the detection of Ureaplasma diversum.

Science.gov (United States)

Marques, Lucas M; Amorim, Aline T; Martins, Hellen Braga; Rezende, Izadora Souza; Barbosa, Maysa Santos; Lobão, Tassia Neves; Campos, Guilherme B; Timenetsky, Jorge

2013-12-27

Ureaplasma diversum in veterinary studies is an undesirable microbe, which may cause infection in bulls and may result in seminal vesiculitis, balanopostitis, and alterations in spermatozoids, whereas in cows, it may cause placentitis, fetal alveolitis, abortion, and birth of weak calves. U. diversum is released through organic secretions, especially semen, preputial and vaginal mucus, conjunctival secretion, and milk. The aim of the present study was to develop a TaqMan probe, highly sensitive and specific quantitative PCR (qPCR) assay for the detection and quantification of U. diversum from genital swabs of bovines. Primers and probes specific to U. diversum 16S rRNA gene were designed. The specificity, detection limit, intra- and inter-assay variability of qPCR to detect this ureaplasma was compared with the results of the conventional PCR assay (cPCR). Swabs of vaginal mucus from 169 cows were tested. The qPCR assay detected as few as 10 copies of U. diversum and was 100-fold more sensitive than the cPCR. No cross-reactivity with other Mollicutes or eubacteria was observed. U. diversum was detected in 79 swabs (46.42%) by qPCR, while using cPCR it was detected in 42 (25%) samples. The difference in cPCR and qPCR ureaplasma detection between healthy and sick animals was not statistically significant. But the U. diversum load in samples from animals with genital disorders was higher than in healthy animals. The qPCR assay developed herein is highly sensitive and specific for the detection and quantification of U. diversum in vaginal bovine samples. Copyright © 2013. Published by Elsevier B.V.

Enrichment of methylated molecules using enhanced-ice-co-amplification at lower denaturation temperature-PCR (E-ice-COLD-PCR) for the sensitive detection of disease-related hypermethylation.

Science.gov (United States)

Mauger, Florence; Kernaleguen, Magali; Lallemand, Céline; Kristensen, Vessela N; Deleuze, Jean-François; Tost, Jörg

2018-05-01

The detection of specific DNA methylation patterns bears great promise as biomarker for personalized management of cancer patients. Co-amplification at lower denaturation temperature-PCR (COLD-PCR) assays are sensitive methods, but have previously only been able to analyze loss of DNA methylation. Enhanced (E)-ice-COLD-PCR reactions starting from 2 ng of bisulfite-converted DNA were developed to analyze methylation patterns in two promoters with locked nucleic acid (LNA) probes blocking amplification of unmethylated CpGs. The enrichment of methylated molecules was compared to quantitative (q)PCR and quantified using serial dilutions. E-ice-COLD-PCR allowed the multiplexed enrichment and quantification of methylated DNA. Assays were validated in primary breast cancer specimens and circulating cell-free DNA from cancer patients. E-ice-COLD-PCR could prove a useful tool in the context of DNA methylation analysis for personalized medicine.

Development and evaluation of a nested-PCR assay for Senecavirus A diagnosis.

Science.gov (United States)

Feronato, Cesar; Leme, Raquel A; Diniz, Jaqueline A; Agnol, Alais Maria Dall; Alfieri, Alice F; Alfieri, Amauri A

2018-02-01

Senecavirus A (SVA) has been associated with vesicular disease in weaned and adult pigs and with high mortality of newborn piglets. This study aimed to establish a nested-PCR assay for the routine diagnosis of SVA infection. Tissue samples (n = 177) were collected from 37 piglets of 18 pig farms located in four different Brazilian states. For the nested-PCR, a primer set was defined to amplify an internal VP1 fragment of 316 bp of SVA genome. Of the 37 piglets, 15 (40.5%) and 23 (62.2%) were positive for the SVA in the RT-PCR and nested-PCR assays, respectively. The SVA RNA was detected in 61/177 (34.5%) samples with the RT-PCR, while the nested-PCR assay showed 84/177 (47.5%) samples with the virus (p PCR and nested-PCR assays, respectively. Nucleotide sequencing analysis revealed similarities of 98.7-100% among SVA Brazilian strains and of 86.6-98% with SVA strains from other countries. The nested-PCR assay in this study was suitable to recover the SVA RNA in biological specimens, piglets, and/or herds that were considered as negative in the RT-PCR assay, and is proposed for the routine investigation of the SVA infection in piglets, especially when other techniques are not available or when a great number of samples has to be examined.

[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.

PCR detection of Bartonella spp. in the dog

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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.

Cervical cancer and human papillomavirus: Epidemiological evidence and perspectives for prevention Cáncer del cérvix y virus del papiloma humano: evidencia epidemiológica y perspectivas para su prevención

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NUBIA MUÑOZ

1997-07-01

calcula se presentan anualmente en el mundo, corresponde a los países en desarrollo. Actualmente se revisa la evidencia epidemiológica que relaciona al virus del papiloma humano (VPH con el cáncer del cérvix. Se ha concluido que alrededor de 90% de los cánceres de cérvix pueden atribuirse a ciertos tipos de VPH. Así, el VPH 16 representa la mayor proporción (50%, seguido por el VPH 18 (12%, el VPH 45 (8% y el VPH 31 (5%. Las asociaciones con estos tipos de VPH son bastante fuertes y consistentes con razones de momios más allá de 15 en todos los estudios de casos y controles en los países con alto y bajo riesgo de cáncer cervical. No obstante, el VPH no constituye una causa suficiente de esta enfermedad; son necesarios ciertos cofactores para que un porcentaje de infecciones persistentes por VPH logre, en algún momento, progresar y dar lugar al cáncer. Entre ellos están los factores del huésped como los tipos de antígenos de histocompatibilidad y la respuesta inmonológica, las influencias que ejercen las hormonas y otros agentes de transmisión sexual, como por ejemplo la Chlamydia trachomatis. Por otra parte, los resultados de los estudios que se llevaron a cabo en España y en Colombia permiten sostener la hipótesis de que los portadores masculinos de VPH desempeñan un papel importante en el desarrollo del cáncer de cérvix que presentan sus esposas. El reconocimiento del sitio tan destacado que ocupa el VPH en el cáncer cervical ha rebasado en mucho las implicaciones de la prevención primaria y secundaria de este padecimiento. Hoy en día se están desarrollando vacunas terapéuticas y profilácticas contra el VPH y su tipificación se está integrando a los programas de detección en estudios piloto de algunos países desarrollados. En las naciones en desarrollo, los programas de detección convencionales y que cuentan con un buen manejo siguen siendo el mejor enfoque para controlar el cáncer del cérvix hasta que pueda utilizarse una vacuna

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

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

PCR performance of a thermostable heterodimeric archaeal DNA polymerase

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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.

Result Variation and Efficiency Kinetics in Real-Time PCR

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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.

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

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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.

PCR biocompatibility of Lab-on-a-chip and MEMS materials

DEFF Research Database (Denmark)

Christensen, Troels Balmer; Pedersen, Christian Møller; Grøndahl, K. G.

2007-01-01

the possibility of interaction between the surfaces and ingredients in the PCR mixture. By proper surface treatment the PCR reaction can be facilitated and in this paper we present a systematic and quantitative study of the impact on the PCR compatibility of a chemical and a biological surface treatment....... The chemical treatments are based on the silanizing agent dichlordimethylsilane [(CH3)(2)SiCl2

Conhecimento de mulheres de 40 a 60 anos sobre o Papillomavirus Humano

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Caroline Freitas Silveira

2011-01-01

Full Text Available El estudio planteó determinar el conocimiento de mujeres, de 40 - 60 años, sobre el Virus Papiloma Humano (VPH. Se trata de una investigación descriptiva, transversal y cuantitativa efectuada con 30 mujeres que esperaban la realización de la prueba de Papanícclacu, en el Centro de Atención Integral a la Salud de la Mujer, en Uberaba - MG, entre octubre de 2009 a enero de 2010. La edad promedio fue de 48,7 años. Sobre la prueba de Papanicolacu, un 96,7% informó que realizó el procedimiento alguna vez y 40% mencionó que la prueba previene el cáncer de cuello uterino. Sobre el VPH,86,7% de las mujeres conocía la sigla, pero 53,3% no sabía lo qué era VPH. Se concluyó que el conocimiento que estas mujeres tenían sobre la prueba Papanícclacu y la sigla VPH era insuficiente. Por lo tanto, se recomienda reforzar las campañas de información para ese rango de edad específico.

Specific PCR-based detection of Alternaria helianthi

DEFF Research Database (Denmark)

Udayashankar, A.C.; Nayaka, S. Chandra; Archana, B.

2012-01-01

Alternaria helianthi is an important seed-borne pathogenic fungus responsible for blight disease in sunflower. The current detection methods, which are based on culture and morphological identification, are time-consuming, laborious and are not always reliable. A PCR-based diagnostic method...... tested. The detection limit of the PCR method was of 10 pg from template DNA. The primers could also detect the pathogen in infected sunflower seed. This species-specific PCR method provides a quick, simple, powerful and reliable alternative to conventional methods in the detection and identification...

Simultaneous detection of three lily viruses using Triplex IC-RT-PCR.

Science.gov (United States)

Zhang, Yubao; Wang, Yajun; Xie, Zhongkui; Yang, Guo; Guo, Zhihong; Wang, Le

2017-11-01

Viruses commonly infecting lily (Lilium spp.) include: Lily symptomless virus (LSV), Cucumber mosaic virus (CMV) and Lily mottle virus (LMoV). These viruses usually co-infect lilies causing severe economic losses in terms of quantity and quality of flower and bulb production around the world. Reliable and precise detection systems need to be developed for virus identification. We describe the development of a triplex immunocapture (IC) reverse transcription (RT) polymerase chain reaction (PCR) assay for the simultaneous detection of LSV, CMV and LMoV. The triplex IC-RT-PCR was compared with a quadruplex RT-PCR assay. Relative to the quadruplex RT-PCR, the specificity of the triplex IC-RT-PCR system for LSV, CMV and LMoV was 100% for field samples. The sensitivity of the triplex IC-RT-PCR system was 99.4%, 81.4% and 98.7% for LSV, CMV and LMoV, respectively. Agreement (κ) between the results obtained from the two tests was 0.968, 0.844 and 0.984 for LSV, CMV and LMoV, respectively. This is the first report of the simultaneous detection of LSV, CMV and LMoV in a triplex IC-RT-PCR assay. In particular we believe this convenient and reliable triplex IC-RT-PCR method could be used routinely for large-scale field surveys or crop health monitoring of lily. Copyright © 2017. Published by Elsevier B.V.

Diseño e implementación de un sistema de seguridad vehicular mediante reconocimiento facial a través de visión artificial

OpenAIRE

Cajas Idrovo, Marco Vinicio; Viri Ávila, Pablo Andrés

2017-01-01

El documento consiste en un sistema de seguridad antirrobo de vehículos basado en visión artificial mediante varias técnicas de reconocimiento facial, el cual permite el encendido y la conducción de personas autorizadas previamente establecida en una base de datos y niega el acceso a otras personas haciendo sonar alarmas, esto se logra mediante la activación del relé de la bomba de gasolina, el reconocimiento se lo hace cada cierto tiempo para a cada momento verificar la identidad de la perso...

Evaluación genotóxica, mediante la prueba de micronúcleos, de la exposición a drogas psicoactivas en individuos del suroccidente colombiano

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LS. Hoyos

2001-07-01

genotóxico de estas drogas mediante la prueba de Micronúcleos (Mn, que identifica fragmentos cromosómicos o cromosomas enteros excluidos del núcleo celular y que es un biomarcador temprano de exposición e indicador de riesgo incrementado de cáncer. El objetivo de este estudio fue: evaluar el daño genético inducido por el consumo de drogas psicoactivas mediante cuantificación de micronúcleos en linfocitos binucleados de sangre periférica de individuos consumidores y no c onsumidores.

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...

"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...

Molecular detection of Toxoplasma gondii in water samples from Scotland and a comparison between the 529bp real-time PCR and ITS1 nested PCR.

Science.gov (United States)

Wells, Beth; Shaw, Hannah; Innocent, Giles; Guido, Stefano; Hotchkiss, Emily; Parigi, Maria; Opsteegh, Marieke; Green, James; Gillespie, Simon; Innes, Elisabeth A; Katzer, Frank

2015-12-15

Waterborne transmission of Toxoplasma gondii is a potential public health risk and there are currently no agreed optimised methods for the recovery, processing and detection of T. gondii oocysts in water samples. In this study modified methods of T. gondii oocyst recovery and DNA extraction were applied to 1427 samples collected from 147 public water supplies throughout Scotland. T. gondii DNA was detected, using real time PCR (qPCR) targeting the 529bp repeat element, in 8.79% of interpretable samples (124 out of 1411 samples). The samples which were positive for T. gondii DNA originated from a third of the sampled water sources. The samples which were positive by qPCR and some of the negative samples were reanalysed using ITS1 nested PCR (nPCR) and results compared. The 529bp qPCR was the more sensitive technique and a full analysis of assay performance, by Bayesian analysis using a Markov Chain Monte Carlo method, was completed which demonstrated the efficacy of this method for the detection of T. gondii in water samples. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Kinetic characterisation of primer mismatches in allele-specific PCR: a quantitative assessment.

Science.gov (United States)

Waterfall, Christy M; Eisenthal, Robert; Cobb, Benjamin D

2002-12-20

A novel method of estimating the kinetic parameters of Taq DNA polymerase during rapid cycle PCR is presented. A model was constructed using a simplified sigmoid function to represent substrate accumulation during PCR in combination with the general equation describing high substrate inhibition for Michaelis-Menten enzymes. The PCR progress curve was viewed as a series of independent reactions where initial rates were accurately measured for each cycle. Kinetic parameters were obtained for allele-specific PCR (AS-PCR) amplification to examine the effect of mismatches on amplification. A high degree of correlation was obtained providing evidence of substrate inhibition as a major cause of the plateau phase that occurs in the later cycles of PCR.

A standard curve based method for relative real time PCR data processing

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Krause Andreas

2005-03-01

Full Text Available Abstract Background Currently real time PCR is the most precise method by which to measure gene expression. The method generates a large amount of raw numerical data and processing may notably influence final results. The data processing is based either on standard curves or on PCR efficiency assessment. At the moment, the PCR efficiency approach is preferred in relative PCR whilst the standard curve is often used for absolute PCR. However, there are no barriers to employ standard curves for relative PCR. This article provides an implementation of the standard curve method and discusses its advantages and limitations in relative real time PCR. Results We designed a procedure for data processing in relative real time PCR. The procedure completely avoids PCR efficiency assessment, minimizes operator involvement and provides a statistical assessment of intra-assay variation. The procedure includes the following steps. (I Noise is filtered from raw fluorescence readings by smoothing, baseline subtraction and amplitude normalization. (II The optimal threshold is selected automatically from regression parameters of the standard curve. (III Crossing points (CPs are derived directly from coordinates of points where the threshold line crosses fluorescence plots obtained after the noise filtering. (IV The means and their variances are calculated for CPs in PCR replicas. (V The final results are derived from the CPs' means. The CPs' variances are traced to results by the law of error propagation. A detailed description and analysis of this data processing is provided. The limitations associated with the use of parametric statistical methods and amplitude normalization are specifically analyzed and found fit to the routine laboratory practice. Different options are discussed for aggregation of data obtained from multiple reference genes. Conclusion A standard curve based procedure for PCR data processing has been compiled and validated. It illustrates that

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

Representación del Conocimiento en curriculo mediante esquemas preconceptuales

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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

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...