Periodic Reporting for period 1 - diaRNAgnosis (A novel platform for the direct profiling of circulating cell-free ribonucleic acids in biofluids)
Reporting period: 2021-01-01 to 2023-05-31
Discovery of ccfRNA Biomarkers: The team validated miRNA-371 as a biomarker for testicular germ cell tumours (TGCTs) and identified potential infertility-related biomarkers. Nine differentially expressed microRNAs were discovered, three of which were confirmed via Real-Time PCR. The team found potential RNA biomarkers for prostate cancer (PCa), including long non-coding RNAs and miRNAs, overexpressed in urine samples from PCa patients.
ODG Platform prototypes: Two prototypes were developed using fluorescence and chemiluminescence protocols. Prototype 1 uses a time-gated detection approach for miRNA detection with photoluminescent-tagged SMART bases, whereas Prototype 2 enhances the chemiluminescent signal with silver and gold nanoparticles. Both performed satisfactorily, but Prototype 2 was selected for future development due to its superior Limit of Detection.
Reagent Development: Reagents were designed and synthesized to enhance the assays on the prototypes and target the newly identified or confirmed ccfRNA biomarkers. A particular success was the synthesis of a SMART base labelled with an Eu(III) luminophore and the successful functionalization of Dynabeads with DGL-Probes. These will be tested with the ODG Platform to investigate further other discovered or validated biomarkers, including long non-coding RNAs.