<p>MicroRNAs (miRNAs) have emerged as robust biomarkers for diagnosing and prognosing gynecological diseases due to their disease-specific expression and remarkable stability in body fluids. Despite the inherent instability of RNA molecules, circulating miRNAs remain well protected through encapsulation in extracellular vesicles (EVs) or binding to RNA-binding proteins and lipoproteins, enabling reliable clinical detection. This review summarizes circulating miRNAs’ biogenesis and clinical relevance, emphasizing their roles in both malignant (e.g., ovarian and endometrial cancers) and benign (e.g., endometriosis and preeclampsia) conditions. Several electrochemical, fluorescent, and colorimetric biosensing platforms have been developed for sensitive and specific miRNA detection. However, most research focuses on cancer-associated miRNAs, leaving those linked to non-malignant diseases largely overlooked. To bridge this gap, this review highlights the need for broader exploration of underrepresented miRNAs and the development of multiplexed biosensing strategies to enhance diagnostic precision. Integrating advanced detection technologies with comprehensive biomarker discovery could significantly improve diagnostic accuracy, expand clinical applications, and accelerate the adoption of miRNA biosensors in point-of-care testing (POCT).</p> Graphical abstract <p>Overview of gynecologic disease biomarkers and their corresponding biosensing detection strategies. The schematic depicts the interrelationship between gynecological diseases, circulating miRNA biomarkers, and biosensor-based detection modalities. The inner circle highlights four representative conditions—ovarian cancer, endometrial cancer, endometriosis, and preeclampsia—whose onset and progression are closely associated with dysregulated serum or plasma miRNAs.</p> <p></p>