Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz

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Gewählte Publikation:

Reinhold, U; Berkin, C; Bosserhoff, AK; Deutschmann, A; Garbe, C; Gläser, R; Hein, R; Krähn, G; Peter, RU; Rappl, G; Schittek, B; Seiter, S; Ugurel, S; Volkenandt, M; Tilgen, W.
Interlaboratory evaluation of a new reverse transcriptase polymerase chain reaction-based enzyme-linked immunosorbent assay for the detection of circulating melanoma cells: a multicenter study of the Dermatologic Cooperative Oncology Group.
J Clin Oncol. 2001; 19(6):1723-1727 [OPEN ACCESS]
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Autor/innen der Med Uni Graz:
Ugurel-Becker Selma

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Purpose: Reverse transcription-polymerase chain reaction (RT-PCR)-based defection of tyrosinase mRNA is the most frequently used laboratory method for the detection of circulating tumor cells in melanoma patients. However, previously published results showed considerable variability in the PCR positivity rates. Materials and Methods: We designed a collaborative study to assess the sensitivity, specificity, and clinical relevance of a new standardized RT-PCR-based enzyme-linked immunosorbent assay (ELISA) for the detection of circulating melanoma cells. Blood samples of healthy donors mixed with cells of a melanoma cell line were prepared in a blinded fashion, and aliquots were sent to seven participating laboratories experienced in RT-PCR. Results: The results demonstrate a high sensitivity(1 melanoma cell/mL blood) and specificity (no false-negatives and 7.4% [2 of 28] false-positives) of the assay and a satisfactory rate of interlaboratory reproducibility. The analysis of aliquots of blinded samples derived from 60 melanoma patients identified tyrosinase mRNA in 17 of 60 (28.3%): three (20%) of 15 stage I patients, two (13.3%) of 15 stage Il patients, five (35.7%) of 14 stage III patients, and seven (43.8%) of 16 stage IV patients. The interlaboratory reproducibility of positive samples, however, was extremely low and indicates the presence of low amounts of targe, mRNA. Conclusion: Reverse transcriptase-PCR ELISA has a high sensitivity and specificity for the detection of tyrosinase mRNA in peripheral blood cells. The low interlaboratory reproducibility for the detection of tumor cells in blood samples of melanoma patients, however, raises the question of relevance of this assay for clinical use. (C) 2001 by American Society of Clinical Oncology.
Find related publications in this database (using NLM MeSH Indexing)
Adult -
Aged -
Aged, 80 and over -
DNA, Neoplasm - analysis
Enzyme-Linked Immunosorbent Assay - methods Enzyme-Linked Immunosorbent Assay - standards
Female -
Humans -
Melanoma - diagnosis Melanoma - pathology
Middle Aged -
Neoplastic Cells, Circulating -
Observer Variation -
Reproducibility of Results -
Reverse Transcriptase Polymerase Chain Reaction - standards
Sensitivity and Specificity -
Skin Neoplasms - diagnosis Skin Neoplasms - pathology

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