Estandarización de un protocolo para aislamiento y amplificación por PCR en tiempo real y o convencional de ADNC de virus de Zika
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Fecha
2016
Autores
Coronell Padilla, Jairo Samir
López Bossio, María Angélica
Trespalacios Cerpa, Cecy Paola
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Ediciones Universidad Simón Bolívar
Facultad de Ciencias de la Salud
Facultad de Ciencias de la Salud
Resumen
El virus del Ztka es una enfermedad transmitida el Vector Aedes aegypti o Aedes
albobictus ( Stegomya aegypti o Stegomya albopicta, nueva clasificación
taxonómica) a un huésped susceptible. Este es un proyecto en el que se trataba
de evidenciar un protocolo para el aislamiento de virus de Zika mediante la
elaboración de la reacción en cadena de polimerasa (PCR) en tiempo real y/o
convencional. Este es un estudio de tipo experimentales, este proyecto contó con
la participación de un biólogo con experiencia en el área de investigación, quien
supervisó todo el proceso, lo cual facilitó ta comprensión de cada paso establecido
en los protocolos seleccionados, para la PCR convencional, y para la
retrotranscripcion se utilizó SuperScript III first-Strand Synthesis System for TRPCR,
y el utilizado para la pcr en tiempo real fue QuantiNova Probe RT-PCR Kit,
guiándonos de los componentes establecidos en cada protocolo y sus respectivas
concentraciones, a la hora de analizar los resuJtados, éstos no son favorables
respecto a lo que se quiere lograr, ya que no se logra la amplificación del virus en
cuestión. Tras los resultados negativos del experimento, se compararon los
procedimientos realizados, con los encontrados en otros estudios similares, para
determinar cuáles son las diferencias, y por ende las posibles causas implicadas
en la no obtención favorable de los objetivos del presente estudio.
Zika virus is a disease the vector Aedes Aedes aegypti or albobictus (Stegomyia aegyptí or Stegomyía albopicta, new taxonomic classification) to a susceptible host. This is a project in which rt was evident a protocol for isolation of Zika virus by devetoping the polymerase chain reaction (PCR) in real time and I or conventional. This is a study of experimental type, this project attended a biologist with experience in the area of research, who oversaw the whole process, which facilitated the understanding of each step established protocols selected for conventional PCR and reverse transcription was used SuperScript III first-Strand Synthesis System for RT-PCR, and used for real-time PCR was QuantiNova Probe RT-PCR Kit, guiding components set for each protocol and their respective concentrations, the when analyzing the resuJts, they are not favorable regarding what is to be achieved, since the amplífication of the virus in question is not achieved. Following the negative results of the experiment, procedures performed, with those found in similar studies, to determine what are the differences, and therefore the possible causes unfavorable involved in achieving the objectives of this study were compared.
Zika virus is a disease the vector Aedes Aedes aegypti or albobictus (Stegomyia aegyptí or Stegomyía albopicta, new taxonomic classification) to a susceptible host. This is a project in which rt was evident a protocol for isolation of Zika virus by devetoping the polymerase chain reaction (PCR) in real time and I or conventional. This is a study of experimental type, this project attended a biologist with experience in the area of research, who oversaw the whole process, which facilitated the understanding of each step established protocols selected for conventional PCR and reverse transcription was used SuperScript III first-Strand Synthesis System for RT-PCR, and used for real-time PCR was QuantiNova Probe RT-PCR Kit, guiding components set for each protocol and their respective concentrations, the when analyzing the resuJts, they are not favorable regarding what is to be achieved, since the amplífication of the virus in question is not achieved. Following the negative results of the experiment, procedures performed, with those found in similar studies, to determine what are the differences, and therefore the possible causes unfavorable involved in achieving the objectives of this study were compared.
Descripción
Palabras clave
Virus, PCR, Zika, Protocolo, Protocol
