G.B. and P.D. contributed equally to this work
Highly specific detection of prostate-specific antigen-positive cells in the blood of patients with prostate cancer or benign prostatic hyperplasia, using a real-time reverse-transcription-polymerase chain reaction method with improved sensitivity
Article first published online: 28 JUN 2008
© 2008 BJU INTERNATIONAL. NO CLAIM TO ORIGINAL US GOVERNMENT WORKS
Volume 102, Issue 11, pages 1566–1572, December 2008
How to Cite
Barbero, G., Destefanis, P., Procida, S., Mandili, G., Ulliers, D., Ceruti, C., Fiori, C., Maule, M. M., Fontana, D., Giribaldi, G. and Turrini, F. (2008), Highly specific detection of prostate-specific antigen-positive cells in the blood of patients with prostate cancer or benign prostatic hyperplasia, using a real-time reverse-transcription-polymerase chain reaction method with improved sensitivity. BJU International, 102: 1566–1572. doi: 10.1111/j.1464-410X.2008.07797.x
- Issue published online: 21 NOV 2008
- Article first published online: 28 JUN 2008
- Accepted for publication 27 March 2008
- prostate-specific antigen;
- quantitative RT-PCR;
- prostate cancer;
- circulating cells;
To assess the presence of circulating prostate-specific antigen (PSA)-expressing cells in patients with prostate cancer or benign prostatic hyperplasia (BPH), and to determine their diagnostic usefulness using a highly sensitive quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) method.
PATIENTS, SUBJECTS AND METHODS
Venous blood samples were obtained from 175 patients with prostate cancer (12 metastatic and 163 not metastatic), 49 with BPH, and 50 healthy volunteers. To improve the specificity and sensitivity of the qRT-PCR three innovative features were combined; a primer overlapping two adjacent exons to inhibit nonspecific amplification; a no-end-point first round amplification to increase the sensitivity; and a target-specific primer for the RT phase to increase the specificity.
The sensitivity of the method was 1 cell/mL of blood and the interassay coefficient of variation was 10.5%. None of the healthy subjects tested positively, while 9% of those with prostatic cancer and 14% with BPH had PSA-positive cells in the blood. There was a positive association between a positive test and the National Comprehensive Cancer Network classification in the patients with newly diagnosed prostate cancer (P = 0.022). There were no additional statistically significant associations.
Our results strongly indicate that although there were no false-positive results and the sensitivity of the method was increased to maximal levels, a low frequency of positive results in patients with prostatic cancer and a high frequency of positive results in those with BPH seems to discourage the use of PSA-positive circulating cells in the search for a clinical diagnosis of prostate cancer.