The advancement of biosensor technology is driving the development of ultrasensitive detection methods, with single-molecule sensing emerging as a particularly promising approach. Nanopore sensing enables the detection of biomarkers at the single-molecule level, offering high sensitivity and specificity for biological sample analysis. In this study, we designed a reconfigurable DNA origami hinge as a molecular carrier for nanopore-based detection of miRNA-141-3p, a biomarker associated with prostate cancer. The hinge undergoes a controlled transition from a closed to an extended state upon binding to miRNA-141-3p, with distinct ionic current signatures recorded before and after reconfiguration during nanopore translocation experiments. Our approach demonstrates detection sensitivity down to the nanomolar level and exhibits excellent selectivity, effectively distinguishing miRNA-141-3p from closely related sequences. These findings highlight the potential of DNA nanostructure-based nanopore sensing for ultrasensitive and selective biomarker detection in biological samples.