The development of simple, robust, and reliable microRNA (miRNA) detection methods is of great significance in the studies of the biological function of miRNAs, molecular diagnostics, treatment of diseases, and targeted drugs. In recent years, with the increasing development of miRNA research, lots of novel approaches were developed for the detection of miRNA in terms of sensitivity, specificity, multiplicity, in situ imaging, etc. In particular, nucleic acid amplification-based methods and many detection techniques such as droplet digital PCR (ddPCR), electrochemiluminescence (ECL), surface-enhanced Raman spectroscopy (SERS), and mass spectrometry (MS) have been employed widely for the highly sensitive detection of miRNA. New progress in miRNA detection has accelerated miRNA functional research and clinical diagnostics. In this review, we summarize the recent progress in the development of miRNA detection methods and new applications. This review will provide guidelines for the development of more advanced miRNA detection methods with high sensitivity and specificity, and applicability to biochemical research, disease diagnosis and therapy.