A new quantitative real-time PCR method to measure human miRNAs using the PROMER technology

MicroRNAs (miRNAs) are small, non-coding RNA molecules that play a crucial role in regulating gene expression. Dysregulation of miRNAs is associated with various human diseases, including cancer. Accurate quantification of miRNAs in bodily fluids or tissue biopsy samples is essential for their use as biomarkers in tumor diagnosis, yet current methods remain suboptimal. In this study, we introduce a novel quantitative real-time PCR platform based on PROMER technology, which relies on RNA:DNA perfect matching in the primer-template interaction, followed by RNAse H2 cleavage to generate a 3'-hydroxyl group for new DNA synthesis. We selected 16 miRNAs implicated in lung cancer and evaluated the specificity and performance of the platform using synthesized miRNA mimics. Each miRNA could be measured with high specificity and accuracy, even at copy numbers as low as 1-10, while non-template controls exhibited minimal amplification. Additionally, key miRNAs involved in tumor progression, such as miR-210, miR-331, and miR-505, were accurately quantified using minimal amounts of plasma. In summary, we present a new quantitative real-time PCR method for miRNA detection using PROMER technology. This platform successfully measured miRNA mimics and was further applied to quantify miRNA species in plasma samples from cancer patients. Our findings support the potential clinical application of this method for early cancer diagnosis using liquid biopsy samples.