The overall goal of the proposal is to develop an automated data acquisition system for non-invasive quantitative cancer detection, screening, and monitoring, based on highly sensitive low volume real time PCR technique and capillary electrophoresis, apply this system for quantitative analysis of the telomerase activity and gene expression in single cancer cells, and to validate this system on clinical specimens using exfoliated cancer cells in voided urine of prostate cancer patients. Phase I of the project aims at building the bench-top prototype of the detection system and determination of the limits of sensitivity of the proposed instrument for quantitative measurement of telomerase activity and hTERT mRNA expression in comparison with the sensitivity of one of the best commercial real-time PCR instruments, OpticonTM, MJ Research, available at SUNY core facility, and/or ABI PRISM TM7700, PE Applied Biosystems. In Phase II we will develop an automated pilot prototype of the data acquisition system, validate the instrument using exfoliated cancer cells in voided urine specimens of prostate cancer patients, and develop optimal protocols for clinical sample preparation and analysis.
