Isolated ductal fluid sample

Abstract

A sample for diagnosis of breast cancer can be prepared by isolating a ductal fluid sample from one duct of a breast of a patient. The isolated ductal fluid is not mixed with ductal fluid from any other duct of the breast. Generally the target duct is not spontaneously discharging. The isolated ductal fluid sample can be examined to determine the presence or absence of a marker associated with cancer or pre-cancer. An isolated ductal fluid sample not mixed with ductal fluid from any other duct of the breast permits identification of the duct which is diseased and provides increased sensitivity for existing diagnostic and analytic techniques.

What is claimed is:  
     
         1 . A method for preparing a sample for use in diagnosis of breast cancer or pre-cancer comprising: 
 isolating a ductal fluid sample from one duct of a breast of a patient, said isolated ductal fluid not mixed with ductal fluid from any other duct of the breast.    
     
     
         2 . A method as in  claim 1 , further comprising: 
 examining the isolated ductal fluid sample to determine the presence or absence of a marker.    
     
     
         3 . A method as in  claim 1 , wherein the duct from which the ductal fluid is isolated is not spontaneously discharging ductal fluid.  
     
     
         4 . A method as in  claim 2 , wherein the marker is selected from the group consisting of: lysophosphatidic acid, a lysophospholipid, paladin, a portion of palladin, a nucleic acid encoding a polypeptide comprising at least a portion of paladin, Lg, a portion of Lg, a nucleic acid encoding a polypeptide comprising at least a portion of Lg, E2F1, a portion of E2F1, a nucleic acid encoding a polypeptide comprising at least a portion of E2F1, T1A12/mac 25, a portion of T1A12/mac 25, a nucleic acid encoding a polypeptide comprising at least a portion of T1A12/mac 25, MAGUK/ZO-1, a portion of MAGUKIZO-1, a nucleic acid encoding a polypeptide comprising at least a portion of MAGUK/ZO-1, repressor of estrogen receptor activity (REA), a portion of REA, a nucleic acid encoding a polypeptide comprising at least a portion of REA, prothymosin alpha (PTA), a portion of PTA, a nucleic acid encoding a polypeptide comprising at least a portion of PTA, c-raf kinase, a portion of c-raf kinase, a nucleic acid encoding a polypeptide comprising at least a portion of c-raf kinase, CD66a, a portion of CD66a, a nucleic acid encoding a polypeptide comprising at least a portion of CD66a, KL-1, a portion of KL-1, a nucleic acid encoding a polypeptide comprising at least a portion of KL-1, cell adhesion molecule 5.2 (CAM 5.2), a portion of CAM 5.2, a nucleic acid encoding a polypeptide comprising at least a portion of CAM 5.2, leptin, a portion of leptin, a nucleic acid encoding a polypeptide comprising at least a portion of leptin, Bcl-2 gene product, at least a portion of Bcl-2 gene product or polypeptide, a nucleic acid encoding a polypeptide encoding at least a portion of Bcl-2 gene product, nuclear matrix 23(nm23), a portion of nm23, a nucleic acid encoding a polypeptide comprising at least a portion of nm23, an apotosis-related protein, a portion of said protein, a nucleic acid encoding a polypeptide comprising at least a portion of the apoptosis-related protein, lipocalin NGAL, a portion of lipocalin NGAL, a nucleic acid encoding a polypeptide comprising at least a portion of lipocalin NGAL, complement regulatory protein CD46, a portion of CD46, a nucleic acid encoding at least a portion of CD46, complement regulatory protein CD59, a portion of CD59, a nucleic acid encoding at least a portion of CD59, a nucleic acid encoding a portion of an FHIT gene, loss of heterozygosity at an FRA3B site, MRP-1/CD9, a portion of MRP-1/CD9, a nucleic acid encoding at least a portion of MRP-1/CD9, KAI1/CD82, a portion of KAI1/CD82, a nucleic acid encoding at least a portion of KAI1/CD82, a Fibroblast Growth Factor (FGF), a portion of FGF, a nucleic acid encoding a polypeptide comprising at least a portion of an FGF, Vascular Epithelial Growth Factor (VEGF), at least a portion of VEGF, a nucleic acid encoding at least a portion of VEGF, Insulin-like Growth Factor -1 (IGF-1), at least a portion of IGF-1, a nucleic acid encoding at least a portion of IGF-1, tumor amplified kinase STK15 (also BTAK and aurora2), at least a portion of STK15, a nucleic acid encoding a polypeptide comprising at least a portion of STK15, TMS-1, a portion of TMS-1, a nucleic acid encoding a polypeptide comprising at least a portion of TMS-1, maspin, at least a portion of maspin, a nucleic acid encoding a polypeptide comprising at least a portion of maspin, at least a portion of breast cancer associated (BRCA) gene, and at least a portion of a BRCA gene product; CDw60 protein, a portion of CDw60 protein or polypeptide, a nucleic acid encoding at least a portion of a CDw60 protein or polypeptide, mammary expressed enzymes including cytochrome P450s, catechol-Omethyltransferase, epoxide hydrolase, peroxidases, glutathione S-transferases, N-acetyltransferases, or sulfotransferases, a nucleic acid encoding at least a portion of a mammary expressed enzyme, Kallikrein 6 (zyme/protease M/neurosin) protein or polypeptide (hK6), a nucleic acid encoding at least a portion of an hK6 protein or polypeptide; and mammastatin protein or polypeptide, a nucleic acid encoding at least a portion of a mammastatin protein or polypeptide.  
     
     
         5 . A method as in  claim 1 , further comprising: 
 examining the isolated ductal fluid to determine absorption of a molecule by abnormal cells in the fluid.    
     
     
         6 . A method as in  claim 5 , wherein the molecule comprises iodide.  
     
     
         7 . A method as in  claim 1 , further comprising: 
 examining the isolated ductal fluid for a loss of heterozygozity.    
     
     
         8 . A method as in  claim 1 , further comprising 
 examining the isolated ductal fluid for the presence of two or more markers.    
     
     
         9 . A method as in  claim 1 , further comprising examining the isolated ductal fluid for the absence of two or more markers.  
     
     
         10 . A method as in  claim 1 , further comprising 
 examining the ductal fluid for the presence of at least one marker and the absence of at least one marker.    
     
     
         11 . A method as in  claim 1 , further comprising analyzing collected ductal epithelial cells by cytology.  
     
     
         12 . A method as in  claim 9 , wherein the markers are selected from the group consisting of DNA content, p53 gene or gene product, and G-actin or a nucleic acid encoding a polypeptide comprising at least a portion of G-actin.  
     
     
         13 . An isolated ductal fluid sample collected from a breast duct in a breast, said isolated ductal fluid not mixed with ductal fluid from any other breast duct.  
     
     
         14 . An isolated ductal fluid sample as in  claim 13 , wherein the sample comprises a marker for analysis.  
     
     
         15 . An isolated ductal fluid sample as in  claim 14 , wherein the analysis comprises determining the presence or absence of the marker.  
     
     
         16 . An isolated ductal fluid sample as in  claim 13 , a portion of said isolated ductal fluid not spontaneously discharging from the breast duct.  
     
     
         17 . An isolated ductal fluid sample as in  claim 13 , wherein the sample comprises 10 or more ductal epithelial cells.  
     
     
         18 . An isolated ductal fluid sample as in  claim 13 , wherein the sample comprises at least one ductal epithelial cells clump.  
     
     
         19 . An isolated ductal fluid sample as in  claim 18 , wherein the clump comprises 5 or more ductal epithelial cells.  
     
     
         20 . A method for analyzing breast markers or epithelial cells, comprising: 
 determining the presence or absence of a marker in an isolated ductal fluid sample collected from a breast duct in a breast, said isolated ductal fluid not mixed with ductal fluid from any other breast duct.    
     
     
         21 . A method as in  claim 20 , wherein the duct from which the ductal fluid is isolated is not spontaneously discharging ductal fluid.  
     
     
         22 . A method as in  claim 20 , wherein the marker is selected from the group consisting of: lysophosphatidic acid, a lysophospholipid, paladin, a portion of palladin, a nucleic acid encoding a polypeptide comprising at least a portion of paladin, Lg, a portion of Lg, a nucleic acid encoding a polypeptide comprising at least a portion of Lg, E2F1, a portion of E2F1, a nucleic acid encoding a polypeptide comprising at least a portion of E2F1, T1A12/mac 25, a portion of T1A12/mac 25, a nucleic acid encoding a polypeptide comprising at least a portion of T1A12/mac 25, MAGUK/ZO-1, a portion of MAGUK/ZO-1, a nucleic acid encoding a polypeptide comprising at least a portion of MAGUK/ZO-1, repressor of estrogen receptor activity (REA), a portion of REA, a nucleic acid encoding a polypeptide comprising at least a portion of REA, prothymosin alpha (PTA), a portion of PTA, a nucleic acid encoding a polypeptide comprising at least a portion of PTA, c-raf kinase, a portion of c-raf kinase, a nucleic acid encoding a polypeptide comprising at least a portion of c-raf kinase, CD66a, a portion of CD66a, a nucleic acid encoding a polypeptide comprising at least a portion of CD66a, KL-1, a portion of KL-1, a nucleic acid encoding a polypeptide comprising at least a portion of KL-1, cell adhesion molecule 5.2 (CAM 5.2), a portion of CAM 5.2, a nucleic acid encoding a polypeptide comprising at least a portion of CAM 5.2, leptin, a portion of leptin, a nucleic acid encoding a polypeptide comprising at least a portion of leptin, Bcl-2 gene product, at least a portion of Bcl-2 gene product or polypeptide, a nucleic acid encoding a polypeptide encoding at least a portion of Bcl-2 gene product, nuclear matrix 23(nm23), a portion of nm23, a nucleic acid encoding a polypeptide comprising at least a portion of nm23, an apotosis-related protein, a portion of said protein, a nucleic acid encoding a polypeptide comprising at least a portion of the apoptosis-related protein, lipocalin NGAL, a portion of lipocalin NGAL, a nucleic acid encoding a polypeptide comprising at least a portion of lipocalin NGAL, complement regulatory protein CD46, a portion of CD46, a nucleic acid encoding at least a portion of CD46, complement regulatory protein CD59, a portion of CD59, a nucleic acid encoding at least a portion of CD59, a nucleic acid encoding a portion of an FHIT gene, loss of heterozygosity at an FRA3B site, MRP-1/CD9, a portion of MRP-1/CD9, a nucleic acid encoding at least a portion of MRP-1/CD9, KAI1/CD82, a portion of KAI1/CD82, a nucleic acid encoding at least a portion of KAI1/CD82, a Fibroblast Growth Factor (FGF), a portion of FGF, a nucleic acid encoding a polypeptide comprising at least a portion of an FGF, Vascular Epithelial Growth Factor (VEGF), at least a portion of VEGF, a nucleic acid encoding at least a portion of VEGF, Insulin-like Growth Factor -1 (IGF-1), at least a portion of IGF-1, a nucleic acid encoding at least a portion of IGF-1, tumor amplified kinase STK15 (also BTAK and aurora2), at least a portion of STK15, a nucleic acid encoding a polypeptide comprising at least a portion of STK15, TMS-1, a portion of TMS-1, a nucleic acid encoding a polypeptide comprising at least a portion of TMS-1, maspin, at least a portion of maspin, a nucleic acid encoding a polypeptide comprising at least a portion of maspin, at least a portion of breast cancer associated (BRCA) gene, and at least a portion of a BRCA gene product; CDw60 protein, a portion of CDw60 protein or polypeptide, a nucleic acid encoding at least a portion of a CDw60 protein or polypeptide, mammary expressed enzymes including cytochrome P450s, catechol-O-methyltransferase, epoxide hydrolase, peroxidases, glutathione S-transferases, N-acetyltransferases, or sulfotransferases, a nucleic acid encoding at least a portion of a mammary expressed enzyme, Kallikrein 6 (zyme/protease M/neurosin) protein or polypeptide (hK6), a nucleic acid encoding at least a portion of an hK6 protein or polypeptide; and mammastatin protein or polypeptide, a nucleic acid encoding at least a portion of a mammastatin protein or polypeptide.  
     
     
         23 . A method as in  claim 20 , wherein the marker is absorption of a molecule by abnormal cells in the fluid.  
     
     
         24 . A method as in  claim 23 , wherein the molecule comprises iodide.  
     
     
         25 . A method as in  claim 20 , wherein the marker is a loss of heterozygozity.  
     
     
         26 . A method as in  claim 20 , wherein the presence of two or more markers is determined.  
     
     
         27 . A method as in  claim 20 , wherein the absence of two or more markers is determined.  
     
     
         28 . A method as in  claim 20  wherein the presence of at least one marker and the absence of at least one marker is determined.  
     
     
         29 . A method as in  claim 20  wherein the marker determined is cytology of ductal epithelial cells.  
     
     
         30 . A method as in  claim 20 , wherein the marker is selected from the group consisting of DNA content, p53 gene or gene product, and G-actin or a nucleic acid encoding a polypeptide comprising at least a portion of G-actin.