Sentinel Node Biopsies in Melanoma Patients: A Protocol for Accurate, Efficient, and Cost-Effective Analysis by Preselection for Immunohistochemistry on the Basis of Tyr-PCR
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Background: Immunohistochemistry (IHC) of serial sectioning is considered the gold standard for detection of melanoma activity in sentinel node (SN) biopsies. However, this is cost and labor intensive. In contrast, tyrosinase reverse transcription-polymerase chain reaction (RT-PCR) is simple and quick, but it is hampered by its extreme sensitivity. This study was performed to test whether a strategy that combines the two methods, using tyrosinase RT-PCR to preselect nodes for IHC, could be accurate and cost effective.
Methods: In 36 patients, SNs were identified by scintigraphy and patent blue uptake. Of each SN, one cross section was analyzed first by hematoxylin and eosin staining. Next, all nodes were examined by serial sectioning and IHC of one-half and tyrosinase RT-PCR of the other. Before comparison, all results were documented in a blinded manner. Material costs and workload estimates were noted per SN.
Results: Fifty-five SNs were retrieved from the 36 patients. Hematoxylin and eosin staining of the first cross section revealed tumor positivity in 3 patients (6 SN). Tyrosinase RT-PCR was positive in 11 of the remaining 33 patients (19 of 49 SN). Of these same 11 patients, only 5 were shown to have tumor-positive SNs by using IHC on serial sections (7 SN). All these nodes had been positive for tyrosinase on PCR. For IHC, an average of 40 sections were prepared and examined per SN at a cost of $200(U.S.)/SN. In contrast, routine tyrosinase RT-PCR costs $37(U.S.)/SN, and takes 5% of the time necessary for IHC. A strategy including hematoxylin and eosin staining on the first cross section, followed by tyrosinase RT-PCR on half of each negative (half) node, could preselect nodes to be taken through serial sectioning. In these series, such a strategy would have prevented serial sectioning and IHC of 30 SN from 22 patients. Apart from a considerable gain in efficiency, this would have reduced material costs by a minimum of $6000 (U.S.). This iscrepancy would be even higher if work intensity of analysts and pathologists were considered.
Conclusions: In routine analysis of SN biopsies in melanoma patients, tyrosinase RT-PCR can be used effectively to preselect nodes for further IHC of serial sections. This method seems both time and cost effective.
KeywordsMelanoma Sentinel node analysis Tyrosinase RT-PCR Immunohistochemistry
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- 1.Goydos JS, Ravikumar TS, Germino FJ, Yudd A, Bancila E. Minimally invasive staging of patients with melanoma: sentinel lymphadenectomy and detection of the melanoma-specific proteins MART-1 and tyrosinase by reverse transcriptase polymerase chain reaction. J Am Coll Surg 1998;187:182–188.Google Scholar
- 4.Godellas CV, Berman CG, Lyman G, et al. The identification and mapping of melanoma regional nodal metastases: minimally invasive surgery for the diagnosis of nodal metastases. Am Surg 1995;61:97–101.Google Scholar
- 5.Smith B, Selby P, Southgate J, Pittman K, Bradley C, Blair GE. Detection of melanoma cells in peripheral blood by means of reverse transcriptase and polymerase chain reaction. Lancet 1991;338:1227–1229.Google Scholar
- 6.van der Velde-Zimmermann D, Roijers JFM, Bouwens-Rombouts A, et al. Molecular test for the detection of tumor cells in blood and sentinel nodes of melanoma patients. Am J Pathol 1996;149:759–764.Google Scholar
- 7.Battayani Z, Grob JJ, Xerri L, et al. Polymerase chain reaction detection of circulating melanocytes as a prognostic marker in patients with melanoma. Arch Dermatol 1995;131:443–447.Google Scholar
- 8.Foss AJ, Guille MJ, Occleston NL, Hykin PG, Hungerford JL, Lightman S. The detection of melanoma cells in peripheral blood by reverse transcription-polymerase chain reaction. Br J Cancer 1995;72:155–159.Google Scholar
- 9.Joseph E, Messina J, Glass FL, et al. Radioguided surgery for the ultrastaging of the patient with melanoma (see Comments). Cancer J Sci Am 1997;3:341–345.Google Scholar
- 10.Curry BJ, Myers K, Hersey P. Polymerase chain reaction detection of melanoma cells in the circulation: relation to clinical stage, surgical treatment, and recurrence from melanoma. J Clin Oncol 1998;16:1760–1769.Google Scholar
- 11.Keilholz U, Willhauck M, Rimoldi D, et al. Reliability of reverse transcription-polymerase chain reaction (RT-PCR)-based assays for the detection of circulating tumour cells: a quality-assurance initiative of the EORTC Melanoma Cooperative Group. Eur J Cancer 1998;34:750–753.Google Scholar
- 12.Dalerba P, Ricci A, Russo V, et al. High homogeneity of MAGE, BAGE, GAGE, tyrosinase and Melan-A/MART-1 gene expression in clusters of multiple simultaneous metastases of human melanoma: implications for protocol design of therapeutic antigenspecific vaccination strategies. Int J Cancer 1998;77:200–204.Google Scholar
- 13.Farthmann B, Eberle J, Krasagakis K, et al. RT-PCR for tyrosinase-mRNA-positive cells in peripheral blood: evaluation strategy and correlation with known prognostic markers in 123 melanoma patients. J Invest Dermatol 1998;110:263–267.Google Scholar
- 14.Blaheta HJ, Schittek B, Breuninger H, et al. Lymph node micrometastases of cutaneous melanoma: increased sensitivity of molecular diagnosis in comparison to immunohistochemistry. Int J Cancer 1998;79:318–323.Google Scholar