부산대학교 도서관

Simple Summary: A multiplexed optimised bias-based preamplification–digital droplet PCR (OBBPA-ddPCR) diagnostic technique was developed and tested in this study. The method estimates blood-derived cell-free DNA (cfDNA) methylation levels of RASSF1A, MIR129-2, NRIP3, and SOX8 target sequences in blood samples. The study included 90 healthy individuals, 40 benign prostatic hyperplasia patients, and 39 prostate cancer (PCa) patients. The risk scores were developed to further reduce unnecessary prostate biopsies and PCa overdiagnosis. The scores included classical risk factors and methylation data, delivering an improved 70% specificity for BPH patients and 100% sensitivity for clinically significant PCa patients. Current prostate carcinoma (PCa) biomarkers, including total prostate-specific antigen (tPSA), have unsatisfactory diagnostic sensitivity and specificity resulting in overdiagnosis and overtreatment. Previously, we described an optimised bias-based preamplification–digital droplet PCR (OBBPA-ddPCR) technique, which detects tumour DNA in blood-derived cell-free DNA (cfDNA) of cancer patients. The current study investigated the performance of newly developed OBBPA-ddPCR-based biomarkers. Blood plasma samples from healthy individuals (n = 90, controls) and PCa (n = 39) and benign prostatic hyperplasia patients (BPH, n = 40) were analysed. PCa and BPH patients had tPSA values within a diagnostic grey area of 2–15 ng/mL, for whom further diagnostic validation is most crucial. Methylation levels of biomarkers RASSF1A, MIR129-2, NRIP3, and SOX8 were found significantly increased in PCa patients compared to controls. By combining classical PCa risk factors (percentage of free PSA compared to tPSA (QfPSA) and patient's age) with cfDNA-based biomarkers, we developed PCa risk scores with improved sensitivity and specificity compared to established tPSA and QfPSA single-marker analyses. The diagnostic specificity was increased to 70% with 100% sensitivity for clinically significant PCa patients. Thus, prostate biopsies could be avoided for 28 out of 40 BPH patients. In conclusion, the newly developed risk scores may help to confirm the clinical decision and prevent unnecessary prostate biopsy. [ABSTRACT FROM AUTHOR]