Estudio de biopsia muscular por punción para el diagnóstico de miopatías, mediante el uso de técnicas de histoquímica e inmunohistoquímica
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Fecha
2009
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Facultad/escuela
Idioma
es
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Universidad Andrés Bello
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Licencia CC
Licencia CC
Resumen
Las miopatías son todas las enfermedades relacionadas con los músculos, para su diagnóstico existen numerosos exámenes, uno de ellos es el histológico, que es de gran significancia, ya que muchas miopatías cursan con alteraciones morfológicas.
El primer paso para realizar éste examen, es la extracción de una muestra de tejido muscular, la cual se obtiene mediante una biopsia que debe ser manejada y procesada de una manera distinta al procesamiento de una muestra histopatológica de rutina, para que el resultado histológico final sea el óptimo. Luego se deben practicar distintas reacciones histológicas, para así poder diferenciar la miopatía que padece el paciente.
En Chile, este tipo de examen se realiza en muy pocos laboratorios, uno de ellos es del Hospital Dr. Luis Calvo Mackenna, Centro de Referencia Nacional en Patología Muscular, el cual cuenta con los recursos necesarios para realizar el estudio histopatológico, sin embargo, existen etapas que deben ser mejoradas, para poder llegar a un diagnóstico oportuno y confiable. Dichos pasos son: el manejo y procesamiento de la biopsia muscular, y las técnicas de coloración empleadas para su observación microscópica. Para optimizarlas fue necesario, primero, la elaboración de un protocolo de toma y manejo del especímen, luego se estandarizaron las distintas técnicas de coloración, las que se dividen en tres tipos: histológicas de rutina (Hematoxilina & Eosina y Tricrómico de Gomori), histoquímicas (enzimáticas como la NADH, COX, etc., y no enzimáticas como PAS y Oil Red O) e inmunohistoquímicas (Distrofinas, Merosina, etc.); con lo que finalmente se logró la elaboración de un panel diagnóstico, el cual consta de técnicas básicas, que permiten evaluar los rasgos generales de las miopatías, y de técnicas más puntuales, las que permiten hacer una diferenciación más específica de la patología en estudio.
Con estos objetivos se pretendió perfeccionar el estudio de la biopsia muscular, desde su extracción hasta la elaboración del diagnóstico final.
Myopathies are all the diseases associated with muscles, for their diagnosis there are numerous test, one of them is the histologic, which is of great significance, as many morphological changes occur with myopathies. The first step to start this exam, is the extraction of a sample of muscle tissue, which is obtained through a biopsy that should be handled and processed in a different way that is processed in a usual histopathological sample, this way the final histological result is optimal. Then have to practice different histological reactions to be able to differentiate the myopathy that affects the patient. In Chile, this type of test is done in few laboratories, one of them is Dr. Luis Calvo Mackenna Hospital, National Reference Center in Muscular Pathology, which has the necessary resources to carry out a histopathological study, however, there are steps that must be improved to achieve a timely and reliable diagnosis. These steps are: the handling and processing of muscle biopsy and the staining techniques used for microscopic observation. To optimize them it was necessary, first, to develop a protocol to take and handle the specimen, then standardize the different coloring techniques, which are divided into three types: routine histological techniques (hematoxylin & eosin and Gomori's Trichrome), Histochemical techniques (enzymatic such as NADH enzyme, COX, among others, and not enzymatic such as PAS and Oil Red O) and Immunohistochemistry techniques (Dystrophin, Merosin, among others), which finally succeeds in developing a diagnostic panel, which consists in fundamental techniques, to assess the general features of myopathies, and more specific techniques, which allow a more specific differentiation of the pathology under study. With these objectives we intended to improve the study of muscle biopsy from its extraction to the elaboration of the final diagnostic.
Myopathies are all the diseases associated with muscles, for their diagnosis there are numerous test, one of them is the histologic, which is of great significance, as many morphological changes occur with myopathies. The first step to start this exam, is the extraction of a sample of muscle tissue, which is obtained through a biopsy that should be handled and processed in a different way that is processed in a usual histopathological sample, this way the final histological result is optimal. Then have to practice different histological reactions to be able to differentiate the myopathy that affects the patient. In Chile, this type of test is done in few laboratories, one of them is Dr. Luis Calvo Mackenna Hospital, National Reference Center in Muscular Pathology, which has the necessary resources to carry out a histopathological study, however, there are steps that must be improved to achieve a timely and reliable diagnosis. These steps are: the handling and processing of muscle biopsy and the staining techniques used for microscopic observation. To optimize them it was necessary, first, to develop a protocol to take and handle the specimen, then standardize the different coloring techniques, which are divided into three types: routine histological techniques (hematoxylin & eosin and Gomori's Trichrome), Histochemical techniques (enzymatic such as NADH enzyme, COX, among others, and not enzymatic such as PAS and Oil Red O) and Immunohistochemistry techniques (Dystrophin, Merosin, among others), which finally succeeds in developing a diagnostic panel, which consists in fundamental techniques, to assess the general features of myopathies, and more specific techniques, which allow a more specific differentiation of the pathology under study. With these objectives we intended to improve the study of muscle biopsy from its extraction to the elaboration of the final diagnostic.
Notas
Tesis (Tecnólogo médico, especialidad en Morfofisiología y Citodiagnóstico)
Palabras clave
Miopatías, Biopsia Muscular, Enzimática Muscular, Histoquímica Muscular, Inmunohistoquímica Muscular