Systems, Kits, and Tests for Detecting Colorectal Cancer

Abstract

A method of detecting a predisposition to, or the incidence of, colorectal cancer in a fecal sample comprises, in a first step (a), detecting the presence of blood in the fecal sample, wherein detection of the presence of blood is indicative of a predisposition to, or the incidence of, colorectal cancer. The method additionally comprises, in second step (b), detecting an epigenetic modification in the DNA contained within the fecal sample, wherein detection of the epigenetic modification is indicative of a predisposition to, or the incidence of, colorectal cancer. Based upon a positive result obtained in either (a) or (b) or in both (a) and (b) a predisposition to, or the incidence of, colorectal cancer is detected. Related methods and kits involve detecting an epigenetic modification in a number of specific genes.

1 .- 81 . (canceled) 
     
     
         82 . A combination of systems comprising:
 (a) a system comprising one or more components for detecting hemoglobin present in a fecal sample, and   (b) a system comprising one or more components for detecting methylation in the promoter of NDRG4 in DNA contained within a fecal sample.   
     
     
         83 . The combination of  claim 82 , wherein the system for detecting hemoglobin present in the fecal sample comprises as a component an antibody that binds to human hemoglobin present in the fecal sample. 
     
     
         84 . The combination of  claim 82 , wherein the system for detecting hemoglobin present in the fecal sample comprises as a component a chromogenic reagent that produces a color change if hemoglobin is present in the fecal sample. 
     
     
         85 . The combination of  claim 82 , wherein the system for detecting methylation in the promoter of NDRG4 in DNA contained within the fecal sample comprises as components primers that hybridize to NDRG4 in DNA contained within the fecal sample after the DNA has been treated with a chemical reagent that selectively modifies either the methylated or non-methylated form of CpG dinucleotide motifs to produce modified DNA in the sample, wherein the primers hybridize to the modified NDRG4 DNA and may be utilized to amplify the modified NDRG4 DNA to produce a detectable amplicon. 
     
     
         86 . The combination of  claim 85 , wherein the system for detecting methylation in the promoter of NDRG4 in DNA contained within the fecal sample further comprises as a component a probe that hybridizes to the detectable amplicon. 
     
     
         87 . The combination of  claim 86 , wherein the primers comprise a forward primer comprising SEQ ID NO:7 and a reverse primer comprising SEQ ID NO:8, and the probe comprises SEQ ID NO:9. 
     
     
         88 . A combination of kits, the combination comprising:
 (a) a kit comprising one or more components for detecting hemoglobin present in a fecal sample, and   (b) a kit comprising one or more components for detecting methylation in the promoter of NDRG4 in DNA contained within a fecal sample.   
     
     
         89 . The combination of  claim 88 , wherein the kit for detecting hemoglobin present in the fecal sample comprises as a component an antibody that binds to human hemoglobin present in the fecal sample. 
     
     
         90 . The combination of  claim 88 , wherein the kit for detecting hemoglobin present in the fecal sample comprises as a component a chromogenic reagent that produces a color change if hemoglobin is present in the fecal sample. 
     
     
         100 . The combination of  claim 88 , wherein the kit for detecting methylation in the promoter of NDRG4 in DNA contained within the fecal sample comprises as components primers that hybridize to NDRG4 in DNA contained within the fecal sample after the DNA has been treated with a chemical reagent that selectively modifies either the methylated or non-methylated form of CpG dinucleotide motifs to produce modified DNA in the sample, wherein the primers hybridize to the modified NDRG4 DNA and may be utilized to amplify the modified NDRG4 DNA to produce a detectable amplicon. 
     
     
         101 . The combination of  claim 100 , wherein the kit for detecting methylation in the promoter of NDRG4 in DNA contained within the fecal sample further comprises as a component a probe that hybridizes to the detectable amplicon. 
     
     
         102 . The combination of  claim 101 , wherein the primers comprise a forward primer comprising SEQ ID NO:7 and a reverse primer comprising SEQ ID NO:8, and the probe comprises SEQ ID NO:9. 
     
     
         103 . A combination of tests comprising:
 (a) a test comprising one or more components for detecting hemoglobin present in a fecal sample, and   (b) a test comprising one or more components for detecting methylation in the promoter of NDRG4 in DNA contained within a fecal sample.   
     
     
         104 . The combination of  claim 103 , wherein the test for detecting hemoglobin present in the fecal sample comprises as a component an antibody that binds to human hemoglobin present in the fecal sample. 
     
     
         105 . The combination of  claim 103 , wherein the test for detecting hemoglobin present in the fecal sample comprises as a component a chromogenic reagent that produces a color change if hemoglobin is present in the fecal sample. 
     
     
         106 . The combination of  claim 103 , wherein the test for detecting methylation in the promoter of NDRG4 in DNA contained within the fecal sample comprises as components primers that hybridize to NDRG4 in DNA contained within the fecal sample after the DNA has been treated with a chemical reagent that selectively modifies either the methylated or non-methylated form of CpG dinucleotide motifs to produce modified DNA in the sample, wherein the primers hybridize to the modified NDRG4 DNA and may be utilized to amplify the modified NDRG4 DNA to produce a detectable amplicon. 
     
     
         107 . The combination of  claim 106 , wherein the test for detecting methylation in the promoter of NDRG4 in DNA contained within the fecal sample further comprises as a component a probe that hybridizes to the detectable amplicon. 
     
     
         108 . The combination of  claim 107 , wherein the primers comprise a forward primer comprising SEQ ID NO:7 and a reverse primer comprising SEQ ID NO:8, and the probe comprises SEQ ID NO:9.