The diaRNAgnosis project has brought together an international consortium of experts to develop a fast, non‑invasive diagnostic tool for two of the most widespread men’s cancers: testicular germ cell tumours (TGCTs) and prostate cancer (PCa). From day one, the partners organised their work into clearly defined stages, ensuring that biological discovery, device engineering and clinical testing progressed in tandem.
In the first phase, scientists focused on finding reliable molecular fingerprints in blood and urine by studying circulating cell-free RNAs (ccfRNAs). Early results confirmed that a molecule called miR-371a-3p is a highly accurate indicator of TGCT. The team also discovered nine additional microRNAs linked to tumour presence—three of which were rigorously validated using real-time PCR—and identified long non-coding RNAs and microRNAs that appear at elevated levels in the urine of men with prostate cancer. These validated markers formed the foundation for the next development steps.
At the same time, scientists at OPTOI and DESTINA Genomica improved the ODG device. This device was realised by merging dynamic chemical labelling (DCL) technology with a silicon photomultiplier (SiPM) detector, further enhancing signal detection and timing resolution. The first version used time-gated fluorescence to detect RNA molecules labelled with special luminescent tags. To reach lower detection limits, a second version shifted to a chemiluminescent approach enhanced by polystyrene nanoparticles, achieving even greater sensitivity. Key assay reagents—including labelled “SMART” nucleotide (SMART-Base) and magnetic beads functionalized with basic peptide nucleic acid (PNA) to capture target RNAs—were developed in parallel, ensuring that the chosen chemiluminescent workflow worked seamlessly.
Midway through the project, two prototype ODG devices were installed in partner laboratories in Trento (Italy) and Granada (Spain). Automated sample handling minimised user variation, and blinded tests on patient-derived samples confirmed that the ODG platform could detect cancer-derived RNAs without any amplification steps, matching or exceeding the performance of standard PCR-based methods. These successful trials demonstrated that the system is ready for broader clinical evaluation.
To support future impact, the consortium also invested in training and outreach. More than fifty researchers and technicians took part in cross‑sector secondments and workshops, gaining hands-on experience in advanced laboratory methods, data analysis and regulatory processes in the fields covered by the project. The team published peer-reviewed articles detailing their discoveries and platform performance and presented results at international conferences. A public-facing website, social-media updates and participation in events such as the European Researchers’ Night helped share the project’s progress with wider audiences.
By the end of the reporting period, diaRNAgnosis had delivered a fully functioning, amplification-free liquid‑biopsy platform, a robust set of RNA biomarkers for TGCT and PCa, and a clear roadmap for regulatory approval and commercialisation. These achievements mark a major step toward easy-to-use tests that could bring faster, more accurate cancer diagnosis into routine clinical practice, ultimately benefiting patients and healthcare systems worldwide.
