We propose to develop an integrated high throughput real-time PCR/capillary electrophoresis (PCR/CE) system for highly sensitive, quantitative detection of molecular markers of cancer, and apply this system for quantitative analysis of the telomerase activity and gene expression in single cancer cells isolated from voided urine of prostate cancer patients. In the proposed system, we will use disposable monolith multi-capillary arrays (MMCA) as a vessel for realtime PCR. The use of MMCA will enable highly sensitive low volume (economical) PCR/CE assays as well as absolute quantification of DNA target in clinical samples based on analysis of terminal dilutions and Poisson statistics. Ultra high sensitivity of the system will be due to the use of single photon detection of fluorescently labeled DNA species. The system operation will be automated and will rely on modern data acquisition, processing, and handling technologies (internet accessibility, standard database approach, etc.) We will carry out the clinical evaluation of the proposed system for non-invasive early detection of prostate cancer, and determine the feasibility of quantitative analysis of telomerase activity for discrimination between benign and malignant hyperproliferation.
