Abstract
INTRODUCTION: MicroRNAs (miRNAs) are small, non-coding RNA molecules critical for cellular function, growth, and development. Recent advances in remote diagnostic technologies have highlighted the potential of urinary miRNAs as non-invasive biomarkers for disease monitoring. This study introduces a simple, rapid, and cost-effective reagent for exosomal miRNA extraction, designed for urine-based exosome screening. We further identified hsa-miR-16-5p (miR-16) as a promising diagnostic biomarker for prostate cancer (PCa), with the goal of integrating this method with biosensors to enable rapid result acquisition and support early cancer detection and treatment planning. MATERIALS AND METHODS: The extraction reagent was formulated using polyethylene glycol (PEG), sodium dodecyl sulfate (SDS), and diethyl pyrocarbonate (DEPC), differing from commercial kits. miRNAs were extracted and validated through RT-qPCR, focusing on miR-16 and the reference control miR-21. Results were compared against commercial kits. A cohort of 39 clinical samples-28 PCa patients, 1 benign prostatic hyperplasia (BPH) case, and 10 healthy controls (Ctr)-was analyzed. Statistical evaluations included T-tests and receiver operating characteristic (ROC) analysis to assess the diagnostic value of miR-16. RESULTS: miR-16 expression significantly differed across PCa stages and between cancerous and non-cancerous individuals (Ctr vs Stage IV: p < 0.05; Stage II vs III: p < 0.05; Stage II vs IV: p < 0.005; Stage III vs IV: p < 0.05). ROC analysis confirmed miR-16's diagnostic potential, particularly in detecting mid-stage PCa. CONCLUSION: The proposed extraction method matches commercial kits in performance but offers notable advantages: a simplified five-step process, reduced extraction time (1.08 hours), and low cost ($0.13/test). These findings support the use of urinary miR-16 as a biomarker for PCa staging and highlight the practical value of the newly developed extraction reagent.