Co-allosteric hairpin probe for detecting microRNA with high specificity

MicroRNAs have emerged as crucial biomarkers for disease diagnosis and prognosis, while their accurate detection remains challenging due to high sequence homology among family members. In this study, a co-allosteric hairpin (CA-HP) probe was designed for highly specific microRNA detection. The CA-HP probe leverages synergistic interactions between the target miRNA and a helper strand to induce a conformational change, resulting in fluorescence activation only upon perfectly matched target recognition. By optimizing the complementary lengths of the target and helper strands, the system achieves significantly enhanced specificity compared to conventional hairpin probes. This strategy offers a robust and facile approach for accurate miRNA profiling, which is crucial for clinical diagnostics and personalized medicine. The CA-HP design also provides a generalizable framework for developing high-specificity nucleic acid probes.