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Optimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells.
McIntyre, I G Spreckley, Katherine Clarke, Robert B Anderson, Elizabeth George, N J
McIntyre, I G
Spreckley, Katherine
Clarke, Robert B
Anderson, Elizabeth
George, N J
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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.
Description
Date
2000-10
Publisher
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From UNPAYWALL
Adobe PDF, 342 KB
Keywords
Cancer Staging
Prostatic Cancer
Prostatic Cancer
Type
Article
Citation
Optimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells. 2000, 83 (8):992-7 Br. J. Cancer