Detection of Micro Metastasis of Melanoma and Breast Cancer in Paraffin-Embedded Tumor Draining Lymph Nodes by Multimarker Quantitative RT-PCR

Abstract

The invention provides a quantitative realtime RT-PCR assay for detection of metastatic breast, gastric, pancreas or colon cancer cells or metastatic melanoma. The assay allows to predict disease recurrence and survival in patients with AJCC stage I and II, and III disease using multimarker panels. The method for detecting metastatic melanoma cells utilizes panels of markers selected from a group consisting of MAGE-A3, GaINAcT, MART-1, PAX3, Mitf, TRP-2, and Tyrosinase. The method for detecting metastatic breast, gastric, pancreas or colon cancer cells in paraffin-embedded samples utilizes panels of markers selected from a group consisting of C-Met, MAGE-A3, Stanniocalcin-1, mammoglobin, HSP27, GaINAcT, CK20, and β-HCG.

1 .- 26 . (canceled) 
     
     
         27 . A method of detecting metastatic melanoma cells in a patient comprising:
 (a) receiving a biological sample from the patient;   (b) amplifying mRNA transcripts encoded by a panel of marker genes, the panel of marker genes comprising GaINAcT, PAX3, MAGE-A3, and optionally MART-1 marker genes, the mRNA transcripts being obtained from nucleic acid from the biological sample; and   (c) quantifying each of the mRNA transcripts encoded by the GaINAcT, PAX3, MAGE-A3, and optionally MART-1 marker genes such that detection of GaINAcT, PAX3, MAGE-A3, and optionally MART-1 indicates presence of occult melanoma in the patient.   
     
     
         28 . The method of  claim 27  wherein the panel further comprises marker genes selected from a group consisting of MITF, TRP-2 and Tyrosinase. 
     
     
         29 . The method of  claim 27  wherein the mRNA transcripts are amplified using real-time reverse transcriptase polymerase chain reaction (qRT-PCR). 
     
     
         30 . The method of  claim 27  wherein the biological sample is selected from a group consisting of paraffin-embedded (PE) melanoma tissues, frozen lymph nodes, and PE lymph nodes. 
     
     
         31 . The method of  claim 27  wherein the biological sample is histopathologically negative for melanoma cells. 
     
     
         32 . The method of  claim 31  wherein histopathology of the biological sample is determined by hematoxylin and eosin staining or immunohistochemistry.