Anemia de Fanconi, Parte 2. Estrategia metodológica para el diagnóstico molecular en pacientes con anemia de Fanconi
Resumen
La anemia de Fanconi es una enfermedad rara, se presenta en 1-5/millon de nacidos vivos. Los pacientes presentan a nivel celular inestabilidad cromosómica, que es la base para su diagnóstico y aunque clínicamente son heterogéneos, hay tres características generales: alteraciones del desarrollo físico, pancitopenia y alto riesgo a desarrollar cáncer. Presenta heterogeneidad genética, ya que se origina por variantes patogénicas en alguno de los 22 genes de la vía FA/BRCA, 20 de estos genes se heredan de manera autosómica recesiva, uno autosómica dominante y uno ligada al X. Debido a esta heterogeneidad, el diagnóstico molecular es complicado, por lo que se necesita una estrategia con varias metodologías. El primer abordaje es la detección de deleciones largas con el ensayo de amplificación de sondas dependiente de ligandos múltiples (MLPA) en los genes FANCA, FANCD2, FANCN/PALB2 y FANCB. Los casos no resueltos por MLPA se canalizan a secuenciación de nueva generación, ya sea por panel dirigido (16 genes FANC), o por secuenciación del exoma completo, finalmente si todavía tenemos pacientes sin genotipo realizamos microarreglos de alta resolución, que constan de sondas a lo largo del genoma para detectar polimorfismos de un solo nucleótido y variaciones en el número de copias, para la búsqueda de grandes deleciones o duplicaciones en los genes FANC, así como para la detección de regiones con homocigosidad, con el propósito de encontrar alelos homocigotos. En este artículo, presentamos la estrategia detallada para realizar la genotipificación de los pacientes AF mexicanos, con un porcentaje de éxito del 80%.
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