Method for the rapid and ultra-sensitive detection of leukemic cells

Abstract

An improved method is disclosed for diagnosing the presence of a chromosomal translocation characteristic of acute myelogenous leukemia. Nucleic acid molecules that may be used in this improved method are described.

What is claimed is:  
     
         1 . A method for diagnosing the presence of a leukemic cell in a sample comprising the steps: 
 A) incubating said sample in the presence of at least one nucleic acid molecule that is capable of hybridizing to: 
 (1) a nucleotide sequence found in chromosome 8 of a non-leukemic human cell or  
 (2) a nucleotide sequence found in chromosome 21 of a non-leukemic human cell; and  
   B) diagnosing said presence by determining whether said sample contains a nucleic acid molecule, or is capable of forming a single cDNA molecule, that is capable of hybridizing to both a nucleotide sequence found in chromosome 8 of a non-leukemic human cell and a nucleotide sequence found in chromosome 21 of a non-leukemic human cell.    
     
     
         2 . The method of  claim 1 , wherein a single nucleic acid molecule is employed in step A.  
     
     
         3 . The method of  claim 2 , wherein said single nucleic acid is substantially incapable of stably hybridizing to either chromosome 8 or chromosome 21 of a non-leukemic cell, but is capable of stably hybridizing to a chromosome 8-chromosome 21 fusion, or to a cDNA produced from said fusion.  
     
     
         4 . The method of  claim 1 , wherein at least two different nucleic acid molecules are employed in step A.  
     
     
         5 . The method of  claim 4 , wherein said at least two different nucleic acid molecules employed in step A are detectably labelled, and wherein said determination is accomplished by in situ hybridization.  
     
     
         6 . The method of  claim 4 , wherein said at least two different nucleic acid molecules are primers, and wherein said determination is accomplished by a primer-mediated amplification of a sequence corresponding to a chromosome 8-chromosome 21 fusion, or to a sequence corresponding to a cDNA produced from said fusion.  
     
     
         7 . The method of  claim 6 , wherein said amplification employs a polymerase chain reaction.  
     
     
         8 . The method of  claim 7 , wherein said amplification employs a nested polymerase chain reaction.  
     
     
         9 . The method of  claim 1 , wherein at least one of said different nucleic acid molecules is selected from the group consisting of SEQ ID NOS: 1 and 2.  
     
     
         10 . A composition of matter comprising: 
 (1) a nucleic acid molecule having a nucleotide sequence found in chromosome 8 of a non-leukemic human cell and    (2) a nucleic acid molecule having a nucleotide sequence found in chromosome 21 of a non-leukemic human cell;    wherein said nucleic acid molecules are capable of hybridizing to a chromosome at respective sites that flank a chromosome 8-chromosome 21 translocation characteristic of acute myelogenous leukemia.    
     
     
         11 . The composition of  claim 10  wherein at least one of said nucleotide sequences is hybridized to its respective site.  
     
     
         12 . The composition of  claim 10  that comprises SEQ ID NOS: 1 and 2.  
     
     
         13 . The composition of  claim 10  wherein at least one of said nucleic acid molecules is detectably labelled.