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. 2000 Oct;83(8):992–997. doi: 10.1054/bjoc.2000.1417

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

I G McIntyre 1,3, K Spreckley 2, R B Clarke 2, E Anderson 2, N W Clarke 2,3, N J R George 1
PMCID: PMC2363566  PMID: 10993644

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. © 2000 Cancer Research Campaign

Keywords: prostate cancer, reverse transcriptase polymerase chain reaction, micrometastasis

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Selected References

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