Purpose: There have been several studies on the presence of circulating tumor cells in the peripheral blood of patients with malignant tumors including prostate cancer (PCa) using reverse transcription-PCR (RT-PCR). One of the aims of these studies was to obtain high sensitivity that would enable early-stage diagnosis. However, they varied in their detection rates, and the methods were rather complicated. We have improved the RT-PCR assay combining three prostate-associated molecules, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and prostate stem cell antigen (PSCA) to reveal patients with poor prognosis. Experimental Design: Peripheral blood samples were obtained from 129 patients including 58 cases of PCa and 71 cases of nonmalignant disorders. Total RNA was extracted from 1 ml of whole blood using a commercially available kit. Results: The sensitivity of PSA-, PSMA-, and PSCA-nested RT-PCR was verified with positive signals of a single LNCaP cell in 1 ml of female blood sample. PSA-, PSMA-, and PSCA-mRNA were detected in 7 (12.1%), 12 (20.7%), and 8 (13.8%) PCa, and in 1, 2, and 0 samples in nonmalignant disorders, respectively. Among 58 PCa patients, each PCR indicated the prognostic value in the hierarchy of PSCA&gt
PSA&gt
PSMA RT-PCR, and extraprostatic cases with positive PSCA PCR indicated lower disease-progression-free survival than those with negative PSCA PCR. Conclusions: The present findings suggest that PSCA PCR would be most promising for the molecular staging of PCa. The present RT-PCR is a highly cost-effective and rapid procedure, enabling the molecular staging of PCa with RT PCR as a diagnostic routine.