Comparison between three molecular methods for detection of blood melanoma tyrosinase mRNA. Correlation with melanoma stages and S100B, LDH, NSE biochemical markers

Clin Chim Acta. 2005 Dec;362(1-2):85-93. doi: 10.1016/j.cccn.2005.05.031. Epub 2005 Jul 27.


Background: The molecular monitoring of circulating tumor cells by reverse transcriptase-PCR (RT-PCR) for patients with melanoma, is still under debate. It may be affected by: a) pre-analytical variability, b) frequency of melanoma-associated gene transcripts and c) the reliability of the methods employed. Few commercial methods are available for the detection of tyrosinase mRNA in blood.

Objective: Comparison between two RT-PCR-nested methods with a third one based on real-time methodology, for detection and quantitation of tyrosinase transcripts, respectively.

Methods: Sixty-two melanoma patients with different AJCC stages and 20 healthy subjects were enrolled. All blood samples were extracted in duplicate with two different methods. Two nested-PCR methods (one commercial and one in house) plus a real time commercial kit were employed.

Results: The two nested PCR methods employed were overimposable, specific and sensitive, at least in the stage III, where there was a concordance between sentinel lymph nodes detection and blood tyrosinase positivity. The different extraction methods did not affect the quality of results, while the commercial real-time kit cannot be used.

Conclusion: Tyrosinase mRNA detection may be therefore employed to monitor the melanoma patients over time in function of response to therapy.

Publication types

  • Comparative Study

MeSH terms

  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / metabolism
  • Female
  • Humans
  • L-Lactate Dehydrogenase / metabolism*
  • Male
  • Melanoma / blood
  • Melanoma / genetics
  • Melanoma / metabolism*
  • Melanoma / pathology*
  • Monophenol Monooxygenase / genetics*
  • Neoplasm Staging
  • Nerve Growth Factors / genetics
  • Nerve Growth Factors / metabolism*
  • Phosphopyruvate Hydratase / genetics
  • Phosphopyruvate Hydratase / metabolism*
  • RNA, Messenger / blood*
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • S100 Calcium Binding Protein beta Subunit
  • S100 Proteins / genetics
  • S100 Proteins / metabolism*
  • Sensitivity and Specificity


  • Biomarkers, Tumor
  • Nerve Growth Factors
  • RNA, Messenger
  • S100 Calcium Binding Protein beta Subunit
  • S100 Proteins
  • S100B protein, human
  • L-Lactate Dehydrogenase
  • Monophenol Monooxygenase
  • Phosphopyruvate Hydratase