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

2.50
Hdl Handle:
http://hdl.handle.net/10541/86671
Title:
Optimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells.
Authors:
McIntyre, I G; Spreckley, Katherine; Clarke, Robert B; Anderson, Elizabeth; Clarke, Noel W ( 0000-0001-7776-8059 ) ; George, N J
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.
Affiliation:
Department of Urology, Withington Hospital, Manchester, M21 1AH, UK.
Citation:
Optimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells. 2000, 83 (8):992-7 Br. J. Cancer
Journal:
British Journal of Cancer
Issue Date:
Oct-2000
URI:
http://hdl.handle.net/10541/86671
DOI:
10.1054/bjoc.2000.1417
PubMed ID:
10993644
Type:
Article
Language:
en
ISSN:
0007-0920
Appears in Collections:
All Christie Publications ; All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorMcIntyre, I Gen
dc.contributor.authorSpreckley, Katherineen
dc.contributor.authorClarke, Robert Ben
dc.contributor.authorAnderson, Elizabethen
dc.contributor.authorClarke, Noel Wen
dc.contributor.authorGeorge, N Jen
dc.date.accessioned2009-11-23T12:12:19Z-
dc.date.available2009-11-23T12:12:19Z-
dc.date.issued2000-10-
dc.identifier.citationOptimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells. 2000, 83 (8):992-7 Br. J. Canceren
dc.identifier.issn0007-0920-
dc.identifier.pmid10993644-
dc.identifier.doi10.1054/bjoc.2000.1417-
dc.identifier.urihttp://hdl.handle.net/10541/86671-
dc.description.abstractThe 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.en
dc.language.isoenen
dc.subjectCancer Stagingen
dc.subjectProstatic Canceren
dc.subject.meshFalse Negative Reactions-
dc.subject.meshFemale-
dc.subject.meshHumans-
dc.subject.meshMale-
dc.subject.meshNeoplasm Staging-
dc.subject.meshPolymerase Chain Reaction-
dc.subject.meshProstate-Specific Antigen-
dc.subject.meshProstatic Neoplasms-
dc.subject.meshRNA, Messenger-
dc.subject.meshReference Values-
dc.subject.meshReproducibility of Results-
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction-
dc.subject.meshSensitivity and Specificity-
dc.titleOptimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells.en
dc.typeArticleen
dc.contributor.departmentDepartment of Urology, Withington Hospital, Manchester, M21 1AH, UK.en
dc.identifier.journalBritish Journal of Canceren
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