The association of the results reverse transcription-PCR (RT-PCR) assays for prostate-specific antigen (PSA) with prostate cancer stage and progression remains controversial. We have previously shown that RT-PCR amplification of a native human glandular kallikrein fragment (hK2-L), but not a splice-variant hK2 transcript (hK2-U), performed preoperatively on peripheral blood before radical prostatectomy is an independent predictor of metastases to lymph nodes [K. M. Slawin, et al., Cancer Res., 60: 7142-7148, 2000]. We evaluated the ability of preoperative peripheral blood RT-PCR for PSA and hK2 to predict prostate cancer progression after surgery. The study group consisted of 224 consecutive patients who underwent radical prostatectomy for clinically localized disease (median follow-up: 52.9 months). Also evaluated were 14 healthy men and 8 patients with hormone-naïve, prostate cancer metastatic to bone. RT-PCR for PSA, hK2-L, and hK2-U were positive in 24, 25, and 26%, respectively, of prostatectomy patients; 88, 71, and 86%, respectively, of patients with metastases to bones; 7, 14, and 36%, respectively, of healthy men. Preoperative RT-PCR-hK2-L positivity was associated with higher final Gleason sum (P = 0.028), with an increased risk of metastases to lymph nodes (P = 0.019) and with overall and aggressive disease progression (P = 0.0231 and P = 0.028, respectively). Preoperative blood RT-PCR-PSA and RT-PCR-hK2-U were not associated with characteristics or outcomes of prostate cancer. Preoperative blood RT-PCR for PSA and hK2-U are neither therapy-guiding staging tools nor prognostic indicators in patients with clinically localized prostate cancer. In contrast, preoperative RT-PCR-hK2-L status is associated with established markers of aggressive prostate cancer.