131 BIOBANKING OF TISSUE OBTAINED FROM ROBOTIC-ASSISTED RADICAL PROSTATECTOMY CAN BE SUCCESSFULLY IMPLEMENTED WITHOUT COMPROMISING RNA INTEGRITY

Abstract

cific antigen (PSA) and cytokeratin 18 (CK18) genes was measured by quantitative RT/PCR, and we have optimized this method by using SABiosciences qPCR Master Mix. We also employ an additional technique of immunomagnetic precipitation to extract RNA from CTCs. Specifically, we coat Dynabeads® (Invitrogen) with equal amounts of antibody to epithelial cell adhesion molecule (EpCAM) and to prostate specific membrane antigen (PSMA). RESULTS: Using a specific primer/probe set, we demonstrate that our qRT/PCR assay is linear over a wide range, and that we can detect PSA transcript in RNA from 1 LNCaP cell. When LNCaP cells are spiked into peripheral blood mononuclear cells (PBMNCs) from men without PCa, we demonstrate that we can detect 1 human prostate cell per 10 7 PBMNCs. In our phase II clinical trial we successfully detected CTCs in 13/17 (76.5%) patients. Our technique of dual antigen pull-down can detect 1 CTC per ml of whole blood of PCa patients. PSA transcripts have not been detected in negative control blood. CONCLUSIONS: We have developed a novel, rapid and sensitive assay suitable for the detection of CTCs in peripheral blood of PCa patients.