Label-free electrochemical biosensor with magnetic self-assembly constructed via PNA-DNA hybridization process on α-Fe2O3/Fe3O4 nanosheets for APOE ε4 genes ultrasensitive detection

A label-free electrochemical DNA detection strategy based on self-assembled α-Fe₂O₃/Fe₃O₄ nanosheets with PNA-DNA hybridization process was developed for ultrasensitive detection of APOE ε4 gene, one of the most robust genetic risks for Alzheimer’s Disease (AD). In this work, magnetic α-Fe₂O₃/Fe₃O₄ heterogeneous nanosheets were prepared by hydrothermal-calcined reduction method and loaded with Au nanoparticles (AuNPs) on their surfaces. The magnetic α-Fe₂O₃/Fe₃O₄@Au nanocomposites significantly enhanced the electrochemical response as a signal amplification matrix and were able to bind to the magnetic glassy carbon electrode (MGCE) surface by magnetic self-assembly. Moreover, owing to the high specificity and stable binding capacity of PNA with respect to the target DNA, the biosensor not only enabled accurate (the limit of detection was estimated to be 0.147 pM) and rapid detection of the APOE ε4 gene, but also exhibited excellent specificity, stability and regeneration capability. Additional, the satisfactory recoveries were also obtained in real samples of human serum, ranging from 92.83 % to 106.22 % with relative standard deviation (RSD) between 0.25 % and 1.85 %. The results possessed important reference value for exploring the application of DNA biosensor technology in the diagnosis of APOE gene mutation.