Optimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells

Br J Cancer. 2000 Oct;83(8):992-7. doi: 10.1054/bjoc.2000.1417.

Abstract

The reverse transcriptase polymerase chain reaction (RT-PCR) is a sensitive technique that can detect prostate-specific messenger RNA in circulating blood. Many authors have studied the potential of RT-PCR as a staging technique in prostate cancer (PC). Clinical sensitivity and in some cases specificity has been disappointing. Few authors have been able to correlate RT-PCR result with patient stage. We have compared the results of using two different RT-PCR protocols with different sensitivities on blood samples from prostate cancer patients. An 80-amplification-cycle nested primer RT-PCR assay had a detection limit of 10 prostate cells and a 50-cycle RT-PCR could detect 20 cells in 5 ml blood. The 80-cycle assay detected prostate mRNA in four of 10 female samples, whereas the 50-cycle assay detected it in none. There was little difference in the assays' ability to detect prostate mRNA in advanced PC patients. The 50-cycle assay could differentiate between hormone-escaped, stable hormone-treated and untreated localized PC patients, whereas the 80-cycle assay could not. Each blood sample must be assayed several times with RT-PCR to avoid false-negative results and, if this is done, assay specificity can be increased with little effect on clinical sensitivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • False Negative Reactions
  • Female
  • Humans
  • Male
  • Neoplasm Staging
  • Polymerase Chain Reaction / methods*
  • Prostate-Specific Antigen / blood*
  • Prostate-Specific Antigen / genetics
  • Prostatic Neoplasms / blood*
  • Prostatic Neoplasms / pathology*
  • RNA, Messenger / blood*
  • Reference Values
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Sensitivity and Specificity

Substances

  • RNA, Messenger
  • Prostate-Specific Antigen