A Highly Sensitive Methylation Assay for Prostate Cancer Diagnosis.

[PURPOSE] Prostate cancer is a prevalent malignancy among males, necessitating precise diagnosis for effective treatment and prognosis. However, there is a lack of accurate, reliable, and cost-effective methods for precise diagnosis of prostate cancer.

[MATERIALS AND METHODS] The bisulfite-treated DNA was amplified by a blocker strand-assisted methylation-specific PCR method, and the signal was amplified by a guiding strand-assisted enzyme/probe detection system. On this basis, an Optimized DNA Methylation Detection Assay was developed. Fifty-five prostate cancer patients and 24 healthy patients were selected for blood/urine sample testing to evaluate the clinical value of the assay.

[RESULTS] The experimental results showed that the detection limit of the Tri-Component Liquid Biopsy Assay reached 0.002%. Assays for six prostate cancer methylation variants were constructed and finally three sites, GSTP1, ADCY4, and HOXA7, were selected for the design of prostate cancer diagnostic panel. The differences in methylation were statistically significant. Additionally, evaluating this approach on liquid biopsies from prostate cancer patients, we obtained a sensitivity and specificity of 89% and 76% respectively. Meanwhile, the cost of a single test on this platform is about $7.5, and the testing time is only about 5 hours.

[CONCLUSIONS] Here we have successfully developed a highly sensitive methylation assay for prostate cancer diagnosis that features both accuracy, efficiency, and low cost. Combined with the established detection panel, this method can realize accurate and non-invasive early diagnosis of prostate cancer, which substantially augments the pragmatic utility of liquid biopsy.