This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Email this article to a colleague Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Save to my personal folders Download to citation manager Rights & Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Reinhold, U. Articles by Tilgen, W. Search for Related Content PubMed PubMed Citation Articles by Reinhold, U. Articles by Tilgen, W. Social Bookmarking                            What's this?

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

From the Department of Dermatology, The Saarland University Hospital, Homburg/Saar; Department of Pathology, University of Aachen, Aachen; Department of Dermatology, University of Kiel, Kiel; Department of Dermatology, Ludwig-Maximilians-University of Munich; Department of Dermatology, Technical University Munich, Munich; Department of Dermatology, University of Tübingen, Tübingen; and Department of Dermatology, University of Ulm, Ulm, Germany.

Address reprint requests to Uwe Reinhold, MD, Department of Dermatology, The Saarland University Hospital, 66421 Homburg/Saar, Germany; email: uwe.reinhold{at}med-rz.uni-sb.de

PURPOSE: Reverse transcription-polymerase chain reaction (RT-PCR)–based detection 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 II 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 target 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.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				S. J. Hughes, L. Xi, W. E. Gooding, D. J. Cole, M. Mitas, J. Metcalf, R. Bhargava, D. Dabbs, J. Ching, L. Kozma, et al. A Quantitative Reverse Transcription-PCR Assay for Rapid, Automated Analysis of Breast Cancer Sentinel Lymph Nodes J. Mol. Diagn., November 1, 2009; 11(6): 576 - 582. [Abstract] [Full Text] [PDF]

				S. Mocellin, D. Hoon, A. Ambrosi, D. Nitti, and C. R. Rossi The Prognostic Value of Circulating Tumor Cells in Patients with Melanoma: A Systematic Review and Meta-analysis Clin. Cancer Res., August 1, 2006; 12(15): 4605 - 4613. [Abstract] [Full Text] [PDF]

				S. Raja, J. Ching, L. Xi, S. J. Hughes, R. Chang, W. Wong, W. McMillan, W. E. Gooding, K. S. McCarty Jr, M. Chestney, et al. Technology for Automated, Rapid, and Quantitative PCR or Reverse Transcription-PCR Clinical Testing Clin. Chem., May 1, 2005; 51(5): 882 - 890. [Abstract] [Full Text] [PDF]

				U. Keilholz, P. Goldin-Lang, N. E. Bechrakis, N. Max, A. Letsch, A. Schmittel, C. Scheibenbogen, K. Heufelder, A. Eggermont, and E. Thiel Quantitative Detection of Circulating Tumor Cells in Cutaneous and Ocular Melanoma and Quality Assessment by Real-Time Reverse Transcriptase-Polymerase Chain Reaction Clin. Cancer Res., March 1, 2004; 10(5): 1605 - 1612. [Abstract] [Full Text] [PDF]

				A. Ulmer, O. Schmidt-Kittler, J. Fischer, U. Ellwanger, G. Rassner, G. Riethmuller, G. Fierlbeck, and C. A. Klein Immunomagnetic Enrichment, Genomic Characterization, and Prognostic Impact of Circulating Melanoma Cells Clin. Cancer Res., January 15, 2004; 10(2): 531 - 537. [Abstract] [Full Text] [PDF]

				R. A. Wascher, D. L. Morton, C. Kuo, R. M. Elashoff, H.-J. Wang, M. Gerami, and D. S.B. Hoon Molecular Tumor Markers in the Blood: Early Prediction of Disease Outcome in Melanoma Patients Treated With a Melanoma Vaccine J. Clin. Oncol., July 1, 2003; 21(13): 2558 - 2563. [Abstract] [Full Text] [PDF]

				B. Mellado, M. del Carmen Vela, D. Colomer, L. Gutierrez, T. Castel, L. Quinto, M. Fontanillas, N. Reguart, J. M. Domingo-Domenech, C. Montagut, et al. Tyrosinase mRNA in Blood of Patients With Melanoma Treated With Adjuvant Interferon J. Clin. Oncol., October 1, 2002; 20(19): 4032 - 4039. [Abstract] [Full Text] [PDF]

				D. Santini, G. Tonini, B. Vincenzi, R. Murace, G. Ferranti, and A. Baldi Skin Lesions in Melanoma and Kaposi's Sarcoma: Case 1. Detection of Circulating Malignant Cells by RT-PCR in a Case of Cutaneous Melanoma in Complete Regression J. Clin. Oncol., March 1, 2002; 20(5): 1411 - 1412. [Full Text] [PDF]