Droplet digital PCR, a prospective technological approach to quantitative profiling of microRNA

Resumo

MicroRNA is a special type of regulatory molecules modulating gene expression. Circulating microRNAs found in blood and other biological body fluids are now considered as potential biomarkers of human pathology. Quantitative changes of particular microRNAs have been recognized in many oncological diseases and other disorders. A recently developed method of droplet digital PCR (ddPCR) possesses a number of advantages making this method the most suitable for verification and validation of perspective microRNA markers of various human pathologies. These advantages include high accuracy and reproducibility of microRNA quantification as well as possibility of direct high-throughput determination of the absolute number of microRNA copies within a wide dynamic range. The present review considers microRNA biogenesis, the origin of circulating microRNAs, and methods used for their quantification. The special technical features of ddPCR, which make this method especially attractive for studying microRNAs as biomarkers of human pathologies and for basic research devoted to aspects of gene regulation by microRNA molecules, are also discussed.