Method of Detection of Prostate Cancer

Abstract

The present invention provides methods and kits useful for detecting neplasia by measuring the methylation level of biomarkers, especially the promoter region of GSTP1 for the detection of prostate adenocarcinoma.

1 . A kit useful for detecting prostate adenocarcinoma comprising:
 a first container containing a pair of primers for amplification of a promoter region of glutathione-S-transferase (GSTP1),   a second container containing a pair of primers for amplification of a region in a reference gene, and   a third container containing a first and second oligonucleotide probe,   
     wherein the first oligonucleotide probe is specific to the amplification of the promoter region of GSTP1 and the second oligonucleotide probe is specific to the amplification of the region in the reference gene, 
     wherein at least one of the primers for the amplification of the promoter region of GSTP1 or one of the oligonucleotide probes is capable of distinguishing between methylated and unmethylated nucleic acid. 
   
   
       2 . The kit of  claim 1  further comprising a fourth container containing a modifying agent that modifies unmethylated cytosine to produce a converted nucleic acid. 
   
   
       3 . The kit of  claim 1  further comprising an instruction disclosing a methylation ratio indicative of prostate adenocarcinoma. 
   
   
       4 . The kit of  claim 1  further comprising an instruction disclosing that a methylation ratio greater than 5 is indicative of prostate adenocarcinoma. 
   
   
       5 . The kit of  claim 1  further comprising an instruction disclosing that a methylation ratio greater than 10 is indicative of prostate adenocarcinoma. 
   
   
       6 . The kit of  claim 1  further comprising an instruction disclosing that a methylation ratio greater than 50 is indicative of prostate adenocarcinoma. 
   
   
       7 . The kit of  claim 1 , wherein the reference gene is MYOD or ACTB. 
   
   
       8 . The kit of  claim 1 , wherein the region of the reference gene is devoid of CpG dinucleotides. 
   
   
       9 . The kit of  claim 1 , wherein the region of the reference gene contains a first and second primer binding site and an oligonucleotide binding site and wherein the first and second primer binding site and the olionucleotide binding site are devoid of CpG dinucleotides. 
   
   
       10 . The kit of  claim 1 , wherein the primers for amplification of the promoter region of GSTP1 are capable of distinguishing between methylated and unmethylated nucleic acid. 
   
   
       11 . The kit of  claim 1 , wherein the first oligonucleotide probe is capable of distinguishing between methylated and unmethylated nucleic acid. 
   
   
       12 . The kit of  claim 1  further comprising an instruction disclosing that the kit is useful for detecting prostate adenocarcinoma in a prostate tissue sample from a subject. 
   
   
       13 . The kit of  claim 1  further comprising a probe for PSA. 
   
   
       14 . A kit useful for detecting prostate adenocarcinoma comprising:
 a first container containing a pair of primers for amplification of a promoter region of glutathione-S-transferase (GSTP1), wherein the primers are capable of distinguishing between methylated and unmethylated nucleic acid, and   an instruction disclosing that the kit is useful for detecting prostate adenocarcinoma in a bodily fluid sample from a subject and that a methylation level of the promoter region of GSTP1 as determined by conventional polymerase chain reaction using the primers in the first container that is higher than the methylation level of the promoter region of GSTP1 in a normal subject is indicative of prostate adenocarcinoma in the subject with a sensitivity no less than 40%.   
   
   
       15 . The kit of  claim 14 , wherein the sensitivity is no less than 50%. 
   
   
       16 . The kit of  claim 14 , wherein the bodily fluid is serum, plasma, ejaculate, or urine. 
   
   
       17 . The kit of  claim 14 , further comprising
 a second container containing a pair of primers for amplification of a region in a reference gene, and   a third container containing a first and second oligonucleotide probe,   
     wherein the first oligonucleotide probe is specific to the amplification of the promoter region of GSTP1 and the second oligonucleotide probe is specific to the amplification of the region in the reference gene, 
     wherein at least one of the primers for the amplification of the promoter region of GSTP1 and one of the oligonucleotide probes is capable of distinguishing between methylated and unmethylated nucleic acid. 
   
   
       18 . The kit of  claim 17 , wherein the reference gene is MYOD or ACTB. 
   
   
       19 . The kit of  claim 17 , wherein the first oligonucleotide probe is capable of distinguishing between methylated and unmethylated nucleic acid.