title: Polymerase chain reaction and immunohistochemistry frequently detect occult melanoma cells in regional lymph nodes of melanoma patients.
abstract: OBJECTIVE
  To evaluate immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) for melanoma associated antigens (MAA) in detecting occult melanoma cells in lymph nodes which were missed on routine pathology.
  
  
  METHODS
  Occult melanoma cells were sought in 436 lymph nodes from 32 patients with cutaneous melanoma of the lower extremities by immunohistochemistry using the melanoma specific antibody HMB-45. The detection sensitivity of routine histology, immunohistochemistry, and RT-PCR was also compared in 23 lymph node samples from six patients.
  
  
  RESULTS
  Immunohistochemistry showed that 15 of 24 patients (62.5%) who had no detectable metastasis by routine histology had at least one lymph node containing HMB-45 positive cells, mostly seen singly in the medullary sinus. No difference was found in known clinicopathological prognostic factors and recurrence rates between the two groups of patients with and without occult HMB-45 positive cells. RT-PCR analyses showed that the nested PCR for tyrosinase was more sensitive than a combination of single round PCR for five different MAA, including tyrosinase, MART-1/Melan A, Pmel-17, TRP-1, and TRP-2, detecting tyrosinase mRNA in six nodes which were negative by HMB-45 immunohistochemistry.
  
  
  CONCLUSIONS
  Immunohistochemistry + RT-PCR is more sensitive than routine histology in detecting occult melanoma cells in lymph nodes. The nested PCR for tyrosinase should be used in future studies investigating the prognostic significance of such lymph node micrometastases.
authors.affiliation: Department of Dermatology, Kanazawa University School of Medicine, Japan. hattanao@med.kanazawa-u.ac.jp
authors.forename: N
authors.initials: N
authors.lastname: Hatta
authors.forename: M
authors.initials: M
authors.lastname: Takata
authors.forename: K
authors.initials: K
authors.lastname: Takehara
authors.forename: K
authors.initials: K
authors.lastname: Ohara
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis; D017931:DNA Primers; D006801:Humans; D007124:Immunoenzyme Techniques; D007264:Inguinal Canal; D008207:Lymphatic Metastasis; D008545:Melanoma / Q000175:diagnosis* / Q000473:pathology / Q000556:secondary*; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012878:Skin Neoplasms / Q000473:pathology*
chemical_list: D000951:Antigens, Neoplasm; D017931:DNA Primers; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins