To validate the RNA-seq results, we used quantitative RT-PCR (qRT-PCR) analysis to evaluate the two most significant hub genes (EEF2 and RPL7) as potential biomarkers of AD for early diagnosis, three upregulated DEGs (TRIOBP, TAS2R39, and BASP1), and three downregulated DEGs (LDHB, NR1D2, and PDK4). Here, PDK4 is linked to Alzheimer disease.