Prueba piloto para la identificación del Campylobacter jejuni, en heces fecales de pacientes con y sin manifestaciones clínicas de enfermedad diarreica aguda, en una institución prestadora de servicio en salud de la ciudad de Barranquilla, mediante PCR múltiple
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2020
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Ediciones Universidad Simón Bolívar
Facultad de Ciencias Básicas y Biomédicas
Facultad de Ciencias Básicas y Biomédicas
Resumen
Durante los últimos años el Campylobacter es foco de
investigaciones en el mundo por ser reconocida como una carga sustancial para la
salud pública. Estos microorganismos que generan gran interés mundial, debido a
que posee un grado alto de supervivencia; son capaces de sobrevivir en diferentes
tipos de ambientes y condiciones, codifican resistencia a los antibióticos y son
causante de un incremento en la morbimortalidad en los últimos años.
Es causante de la campilobacteriosis que es una zoonosis causada por la bacteria
Campylobacter jejuni manifestándose de forma asintomática o causando enteritis
bacteriana y enfermedad diarreica aguda (EDA) con alteraciones clínicas severas.
Esta bacteria se puede adquirir por el consumo de alimentos y aguas contaminados
y puede pasar desapercibido por falta de un diagnóstico específico.
Objetivos: Identificar Campylobacter jejuni, aislada en heces fecales de pacientes
con y sin manifestaciones clínicas de enfermedades diarreicas agudas en una
institución prestadora de servicio de salud en la ciudad de Barranquilla, mediante
técnica de PCR múltiple.
Materiales y Métodos: Se hizo una prueba piloto donde se recolectaron 9 muestras
de heces fecales de pacientes con y sin manifestaciones clínicas de EDA en una
institución prestadora de servicio en salud en Barranquilla. Para la identificación de
género y especie se hizo mediante métodos microbiológicos convencionales y
métodos moleculares como la Reacción en Cadena de la Polimerasa (PCR). La
extracción del ADN bacteriano se realizó utilizando el kit DNA DNeasy (Quiagen)
utilizando el protocolo descrito por la casa comercial. La PCR Múltiple se realizó en
el Termociclador CFX96, utilizando cebadores específicos para el gen 16S rRNA
para identificar género y el gen cia-B específico para especie de Campylobacter
jejuni. El producto de PCR se analizó mediante electroforesis en gel de agarosa 2%
During the last years, Campylobacter has been the focus of research in the world because it is recognized as a substantial burden on public health. These microorganisms that generate great world interest, because they have a high degree of survival; they are capable of surviving in different types of environments and conditions, they encode resistance to antibiotics and are the cause of an increase in morbidity and mortality in recent years. It is the cause of campylobacteriosis which is a zoonosis caused by the Campylobacter jejuni bacteria manifesting asymptomatically or causing bacterial enteritis and acute diarrheal disease (ADD) with severe clinical alterations. This bacterium can be acquired through the consumption of contaminated food and water and can go unnoticed for lack of a specific diagnosis. Objective: To identify Campylobacter jejuni, isolated in feces from patients with and without clinical manifestations of acute diarrheal diseases in an institution that provides health services in the city of Barranquilla, using multiple PCR technique. Materials and Methods: A pilot test was carried out where 9 stool samples were collected from patients with and without clinical manifestations of ADD, in a health service provider institution in Barranquilla. For the identification of genus and species, it was done using conventional microbiological methods and molecular methods such as the Polymerase Chain Reaction (PCR). The extraction of bacterial DNA was carried out using the DNA DNeasy kit (Quiagen), using the protocol described by the commercial house and Multiplex PCR was performed in the CFX96 Thermal Cycler, using specific primers for the 16S rRNA gene to identify genus and the cia-B gen of Campylobacter jejuni species. The PCR product was analyzed by gel electrophoresis. 2% agarose.
During the last years, Campylobacter has been the focus of research in the world because it is recognized as a substantial burden on public health. These microorganisms that generate great world interest, because they have a high degree of survival; they are capable of surviving in different types of environments and conditions, they encode resistance to antibiotics and are the cause of an increase in morbidity and mortality in recent years. It is the cause of campylobacteriosis which is a zoonosis caused by the Campylobacter jejuni bacteria manifesting asymptomatically or causing bacterial enteritis and acute diarrheal disease (ADD) with severe clinical alterations. This bacterium can be acquired through the consumption of contaminated food and water and can go unnoticed for lack of a specific diagnosis. Objective: To identify Campylobacter jejuni, isolated in feces from patients with and without clinical manifestations of acute diarrheal diseases in an institution that provides health services in the city of Barranquilla, using multiple PCR technique. Materials and Methods: A pilot test was carried out where 9 stool samples were collected from patients with and without clinical manifestations of ADD, in a health service provider institution in Barranquilla. For the identification of genus and species, it was done using conventional microbiological methods and molecular methods such as the Polymerase Chain Reaction (PCR). The extraction of bacterial DNA was carried out using the DNA DNeasy kit (Quiagen), using the protocol described by the commercial house and Multiplex PCR was performed in the CFX96 Thermal Cycler, using specific primers for the 16S rRNA gene to identify genus and the cia-B gen of Campylobacter jejuni species. The PCR product was analyzed by gel electrophoresis. 2% agarose.
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Palabras clave
EDA, Campylobacteriosis, PCR, Zoonosis, Campylobacter jejuni
