Innovative Applications and Challenges of Isothermal Amplification Technology in miRNA Detection.

A class of endogenous non-coding RNAs with a length of roughly 18-25 nucleotides is known as microRNAs (miRNAs). They have been established as the best biomarkers for early cancer diagnosis, molecular subtyping, and prognostic evaluation. They also play important roles in important biological processes like cell proliferation, differentiation, apoptosis, and tumorigenesis through post-transcriptional regulatory networks. Traditional detection technologies (such as quantitative Polymerase Chain Reaction (qPCR), Northern blot, and microarrays) exhibit high sensitivity, yet they rely on expensive thermocycling equipment, involve cumbersome procedures, and are time-consuming, making it difficult to meet the demands of clinical point-of-care testing. In recent years, isothermal amplification technologies, represented by loop-mediated isothermal amplification (LAMP), Rolling Circle Amplification (RCA), Strand Displacement Amplification (SDA), and Exponential Amplification Reaction (EXPAR), have emerged as a research frontier in the field of miRNA detection, by virtue of their advantages including elimination of thermal cycling requirements, fast reaction kinetics, high sensitivity, and simple operation. This study aims to systematically organize the core principles, latest advancements, and integrated innovation patterns of isothermal amplification-based miRNA detection technologies, and provide support for the development of miRNA detection platforms suitable for clinical point-of-care testing.