Cutting-edge RNA quantitative detection methods for rapid and on-site analysis

RNA, a vital biomarker, is crucial for molecular diagnostics, encompassing pathogen detection, gene expression, tumor diagnosis, prognosis, and drug efficacy. Since the COVID-19 outbreak, its role in clinical settings has grown. This review examines established RNA detection methods like qPCR, sequencing, Nanostring, and fluorescence in situ hybridization (FISH), highlighting recent progress and trends. It also focuses on new Point-of-Care Testing (POCT) methods developed over the last five years, categorizing them by the necessity for reverse transcription and nucleic acid amplification. The review emphasizes advances in multiplex detection tailored for POCT, aiming to update researchers and clinicians on the latest RNA detection technologies. Among various RNA detection methods, the amplification-free approach is preferable for POCT applications compared to those requiring reverse transcription and sequence-dependent amplification, as they simplify the detection process by reducing technical complexity while retaining high sensitivity.