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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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Volume 44, Issue 12

Issues

Molecular detection of tyrosinase transcripts in peripheral blood from patients with malignant melanoma: correlation of PCR sensitivity threshold with clinical and pathologic disease characteristics

Georgios Mitropapas
  • Department of Experimental Physiology, Medical School, University of Athens, Goudi, Athens, Greece
  • Other articles by this author:
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/ Andrianos Nezos
  • Department of Experimental Physiology, Medical School, University of Athens, Goudi, Athens, Greece
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/ Antonis Halapas
  • Department of Experimental Physiology, Medical School, University of Athens, Goudi, Athens, Greece
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/ Nikolaos Pissimissis
  • Department of Experimental Physiology, Medical School, University of Athens, Goudi, Athens, Greece
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/ Peter Lembessis
  • Department of Experimental Physiology, Medical School, University of Athens, Goudi, Athens, Greece and Endo/OncoResearch Laboratories, Diagnostic Medical Center, Ampelokipi, Athens, Greece
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Antigone Sourla
  • Department of Experimental Physiology, Medical School, University of Athens, Goudi, Athens, Greece and Endo/OncoResearch Laboratories, Diagnostic Medical Center, Ampelokipi, Athens, Greece
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/ Pericles Vassilopoulos / Michael Koutsilieris
  • Department of Experimental Physiology, Medical School, University of Athens, Goudi, Athens, Greece
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Published Online: 2006-12-13 | DOI: https://doi.org/10.1515/CCLM.2006.260

Abstract

Background: Positive molecular detection of tyrosinase transcripts (TYR mRNA) in RNA extracts of peripheral blood (PB) samples from patients with malignant melanoma provides evidence of disease dissemination.

Methods: Total RNA extracted from PB was quantified and subjected to RT-PCR under ultra-sensitive and reduced-sensitivity PCR conditions using SSRT-II. Positive TYR mRNA detection in 78 melanoma patients and 40 healthy volunteers was correlated with clinical stage, Breslow's evaluation of tumor thickness, Clark's assessment of tumor invasion, the location of the primary tumor site, and tumor histology. The assay sensitivity was evaluated by spiking PB with the melanoma cell line SK-MEL-28.

Results: Using ultra-sensitive PCR conditions, eight out of 40 RNA (20%) samples from healthy volunteers and 50 out of 78 RNA (64.1%) samples from melanoma patients tested positive. Using reduced-sensitivity PCR conditions, we found only two positives in 40 RNA samples from healthy subjects and 20 positives in 78 RNA samples (25.6%) from melanoma patients. Only positive PCR samples for the reduced-sensitivity PCR assay correlated significantly with stage IV (metastatic) disease (p=0.0395). There was no significant correlation between positive TYR mRNA samples for either PCR condition (ultra-sensitive and reduced-sensitivity) with Breslow's classification of tumor thickness, Clark's assessment of tumor invasion, location of the primary tumor site, and type of tumor histology.

Conclusions: We conclude that reduced-sensitivity rather than ultra-sensitive PCR conditions correlate with clinical stage in melanoma patients.

Clin Chem Lab Med 2006;44:1403–9.

Keywords: malignant melanoma; mRNA; peripheral; polymerase chain reaction; tyrosinase

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About the article

Corresponding author: Michael Koutsilieris, MD, PhD, Professor of Experimental Physiology and Chairman of the Departments of Basic Sciences, Medical School, University of Athens, Micras Asias 75, Goudi, 11527, Athens, Greece Phone: +30-210-7462597, Fax: +30-210-74625761,


Received: 2006-06-09

Accepted: 2006-09-12

Published Online: 2006-12-13

Published in Print: 2006-12-01


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 44, Issue 12, Pages 1403–1409, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2006.260.

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