This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zippelius, A. Articles by Pantel, K. Search for Related Content PubMed PubMed Citation Articles by Zippelius, A. Articles by Pantel, K.

Analytical Variables of Reverse Transcription-Polymerase Chain Reaction-based Detection of Disseminated Prostate Cancer Cells¹

Institut für Immunologie der Universität München, Munich, 80336 Germany

Early systemic spread of occult tumor cells that may develop into founders of incurable distant metastasis has been identified in prostate cancer patients by reverse transcription-PCR (RT-PCR) amplification of prostate-specific antigen (PSA) mRNA. Nevertheless, the introduction of this new staging tool into the clinical setting has been hampered by the disparate and contradictory data on the sensitivity and specificity of RT-PCR methods reported recently. We used PSA RT-PCR to examine the influence of analytical variables such as priming and enzyme of reverse transcriptase reaction, temperature and time of primer annealing, primer extension and denaturation, as well as the concentrations of magnesium chloride, Taq polymerase, deoxynucleotide triphosphate, primers and BSA on the amplification process. By systematically varying these chemical and physical components, we could demonstrate a significant increase in amplification yield and in stringency of primer annealing. This may explain the wide variety of published findings on molecular staging of prostate cancer, which currently impedes the clinical introduction of PSA RT-PCR assays in prostate cancer. Methodological analyses are needed for standardization and quality assurance to achieve reproducible molecular methods that can be used in clinical practice.

This article has been cited by other articles:

				G. Giribaldi, S. Procida, D. Ulliers, F. Mannu, R. Volpatto, G. Mandili, L. Fanchini, O. Bertetto, G. Fronda, L. Simula, et al. Specific Detection of Cytokeratin 20-Positive Cells in Blood of Colorectal and Breast Cancer Patients by a High Sensitivity Real-Time Reverse Transcriptase-Polymerase Chain Reaction Method J. Mol. Diagn., February 1, 2006; 8(1): 105 - 112. [Abstract] [Full Text] [PDF]

				S. Braun and B. Naume Circulating and Disseminated Tumor Cells J. Clin. Oncol., March 10, 2005; 23(8): 1623 - 1626. [Full Text] [PDF]

				K. Pantel, V. Muller, M. Auer, N. Nusser, N. Harbeck, and S. Braun Detection and Clinical Implications of Early Systemic Tumor Cell Dissemination in Breast Cancer Clin. Cancer Res., December 15, 2003; 9(17): 6326 - 6334. [Abstract] [Full Text] [PDF]

				D. Schamhart, J. Swinnen, K.-H. Kurth, A. Westerhof, R. Kusters, H. Borchers, and C. Sternberg Numeric Definition of the Clinical Performance of the Nested Reverse Transcription-PCR for Detection of Hematogenous Epithelial Cells and Correction for Specific mRNA of Non-Target Cell Origin as Evaluated for Prostate Cancer Cells Clin. Chem., September 1, 2003; 49(9): 1458 - 1466. [Abstract] [Full Text] [PDF]

				G. Mehes, A. Luegmayr, R. Kornmuller, I. M. Ambros, R. Ladenstein, H. Gadner, and P. F. Ambros Detection of Disseminated Tumor Cells in Neuroblastoma: 3 Log Improvement in Sensitivity by Automatic Immunofluorescence plus FISH (AIPF) Analysis Compared with Classical Bone Marrow Cytology Am. J. Pathol., August 1, 2003; 163(2): 393 - 399. [Abstract] [Full Text] [PDF]

				F A Vlems, J H S Diepstra, I M H A Cornelissen, T J M Ruers, M J L Ligtenberg, C J A Punt, J H J M van Krieken, T. Wobbes, and G N P van Muijen Limitations of cytokeratin 20 RT-PCR to detect disseminated tumour cells in blood and bone marrow of patients with colorectal cancer: expression in controls and downregulation in tumour tissue Mol. Pathol., June 1, 2002; 55(3): 156 - 163. [Abstract] [Full Text] [PDF]

				Y. B. Wetherill, C. E. Petre, K. R. Monk, A. Puga, and K. E. Knudsen The Xenoestrogen Bisphenol A Induces Inappropriate Androgen Receptor Activation and Mitogenesis in Prostatic Adenocarcinoma Cells Mol. Cancer Ther., May 1, 2002; 1(7): 515 - 524. [Abstract] [Full Text] [PDF]