title: Clinical relevance of molecular staging for melanoma: comparison of RT-PCR and immunohistochemistry staining in sentinel lymph nodes of patients with melanoma.
abstract: OBJECTIVE
  To determine the clinical significance of a molecular assay based on the reverse transcriptase polymerase chain reaction (RT-PCR) for the presence of micrometastatic melanoma cells in sentinel lymph nodes (SLNs).
  
  
  BACKGROUND
  Routine histologic examination of lymph nodes often underestimates the presence of micrometastatic disease. The authors have previously shown that an RT-PCR assay designed to detect melanocyte-specific expression of the tyrosinase gene could be used to define a population of patients at higher risk for both recurrence and death compared with routine hematoxylin and eosin (H&E) histology. In this study, the authors used the tyrosinase RT-PCR assay in a patient population examined by a more detailed histologic analysis, including S-100 immunohistochemistry.
  
  
  METHODS
  Patients underwent lymphatic mapping and SLN biopsy. SLN specimens were bivalved, and half of each specimen was serially sectioned and examined by routine H&E histology and S-100 immunohistochemistry. The other half of each specimen was analyzed by a nested RT-PCR assay.
  
  
  RESULTS
  Hematoxylin and eosin histology detected metastatic disease in 36 (16%) of the 233 patients tested. S-100 immunohistochemistry detected micrometastatic disease in another 16 patients, and 114 (63%) of 181 patients with histology-negative nodes had positive findings on RT-PCR. There were significant differences between PCR-positive and PCR-negative patient groups in Breslow thickness, Clark level, and the presence of ulceration of the primary tumor, factors that have been shown to correlate with recurrence and survival.
  
  
  CONCLUSIONS
  These results suggest that RT-PCR can increase the sensitivity of detection of metastatic melanoma cells in SLNs over the current standard methods, including H&E histology and S-100 immunohistochemistry. Further long-term follow-up is needed to detect actual differences in recurrence and overall survival.
authors.affiliation: Cutaneous Oncology Program, H. Lee Moffitt Cancer Center and Research Institute at the University of South Florida, Tampa, Florida 33612, USA.
authors.forename: W
authors.initials: W
authors.lastname: Li
authors.forename: A
authors.initials: A
authors.lastname: Stall
authors.forename: S C
authors.initials: SC
authors.lastname: Shivers
authors.forename: J
authors.initials: J
authors.lastname: Lin
authors.forename: F
authors.initials: F
authors.lastname: Haddad
authors.forename: J
authors.initials: J
authors.lastname: Messina
authors.forename: L F
authors.initials: LF
authors.lastname: Glass
authors.forename: G
authors.initials: G
authors.lastname: Lyman
authors.forename: D S
authors.initials: DS
authors.lastname: Reintgen
mesh_terms: D000328:Adult; D000368:Aged; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000473:pathology*; D008297:Male; D008545:Melanoma / Q000473:pathology*; D008875:Middle Aged; D009367:Neoplasm Staging / Q000379:methods*; D011446:Prospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012878:Skin Neoplasms / Q000473:pathology*