Estandarización de la reacción en cadena de la polimerasa (PCR) específica para la amplificación del segmento rs429358 del gen APOE4 en Alzheimer tardío

Abstract

Alzheimer's disease (AD) is a neurodegenerative disease that affects memory and the ability to generate new memories in people with AD. A relationship has been found between the different alleles encoding the APOE protein and late onset AD. The present work seeks to amplify and standardize the Polymerase Chain Reaction (PCR) specifically the rs429358 polymorphism located in exon 4 of the 19q13.32 locus (APOE4). Standardization of this PCR test is crucial for genetic and clinical studies. The ability to detect and analyze the rs429358 polymorphism efficiently opens new avenues for research and potentially for improving the quality of life of those at risk of developing AD. Methodology: Primers were designed with in silico test runs where amplification and specificity were demonstrated, after which they were sent to INVITROGEN for synthesis. In PCR standardization, we tried to optimize the conditions to ensure specific and reproducible amplification of APOE exon 4. This included adjusting the concentration of reagents and primers, hybridisation temperature, duration and number of cycles. Results: The primers synthesized and tested in silico were not specific for in situ amplification due to their self-complementarity.