Method for the detection of gene transcripts in blood and uses thereof

Abstract

The present invention is directed to detection and measurement of gene transcripts in blood. Specifically provided is a RT-PCR analysis performed on a drop of blood for detecting, diagnosing and monitoring diseases using tissue-specific primers. The present invention also describes methods by which delineation of the sequence and/or quantitation of the expression levels of disease-associated genes allows for an immediate and accurate diagnostic/prognostic test for disease or to assess the effect of a particular treatment regimen.

1 . A method of profiling gene expression in a human subject, the method comprising: determining a level of RNA encoded by a gene in a blood sample of the subject, wherein the gene is identified as KIAA0796, thereby profiling gene expression in a human subject. 
     
     
         2 . The method of  claim 1 , wherein the subject has a disease. 
     
     
         3 . The method of  claim 2 , wherein the disease is colorectal cancer. 
     
     
         4 . The method of  claim 1 , wherein the subject is healthy. 
     
     
         5 . The method of  claim 1 , wherein determining the level is performed using at least one oligonucleotide of predetermined sequence. 
     
     
         6 . The method of  claim 5 , wherein the at least one oligonucleotide is specific for RNA encoded only by the gene in blood of human subjects, and/or is specific for cDNA complementary to RNA encoded only by the gene in blood of human subjects. 
     
     
         7 . The method of  claim 6 , wherein determining the level is performed by amplifying of RNA encoded by the gene to form amplified product, using at least one primer, and quantifying the amplified product, wherein the at least one oligonucleotide comprises the at least one primer. 
     
     
         8 . The method of  claim 6 , wherein determining the level is performed by hybridizing cDNA complementary to RNA encoded by the gene with at least one immobilized probe to form hybridization product, and quantifying the hybridization product, wherein the at least one oligonucleotide comprises the at least one probe. 
     
     
         9 . The method of  claim 1 , wherein determining the level is performed by amplifying RNA encoded by the gene. 
     
     
         10 . The method of  claim 1 , wherein determining the level is performed using an immobilized probe. 
     
     
         11 . The method of  claim 1 , wherein determining the level is performed by quantifying cDNA generated from RNA encoded by said gene. 
     
     
         12 . The method of  claim 1 , wherein determining the level is performed by quantifying EST generated from RNA encoded by the gene. 
     
     
         13 . The method of  claim 1 , wherein the level of RNA encoded by the gene is determined relative to a level of RNA encoded by an internal control gene in the blood sample. 
     
     
         14 . A method of characterizing a body state in a human subject, the method comprising: determining a level of RNA encoded by a gene in a blood sample of the subject, wherein the gene is identified as KIAA0796, thereby characterizing a body state in a human subject. 
     
     
         15 . The method of  claim 14 , wherein the body state is a disease. 
     
     
         16 . The method of  claim 15 , wherein the disease is colorectal cancer. 
     
     
         17 . The method of  claim 14 , wherein the subject is healthy. 
     
     
         18 . The method of  claim 14 , wherein determining the level is performed using at least one oligonucleotide of predetermined sequence. 
     
     
         19 . The method of  claim 18 , wherein the at least one oligonucleotide is specific for RNA encoded only by the gene in blood of human subjects, and/or is specific for cDNA complementary to RNA encoded only by the gene in blood of human subjects. 
     
     
         20 . The method of  claim 19 , wherein determining the level is performed by amplifying of RNA encoded by the gene to form amplified product, using at least one primer, and quantifying the amplified product, wherein the at least one oligonucleotide comprises the at least one primer. 
     
     
         21 . The method of  claim 19 , wherein determining the level is performed by hybridizing cDNA complementary to RNA encoded by the gene with at least one immobilized probe to form hybridization product, and quantifying the hybridization product, wherein the at least one oligonucleotide comprises the at least one probe. 
     
     
         22 . The method of  claim 14 , wherein determining the level is performed by amplifying RNA encoded by the gene. 
     
     
         23 . The method of  claim 14 , wherein determining the level is performed using an immobilized probe. 
     
     
         24 . The method of  claim 14 , wherein determining the level is performed by quantifying cDNA generated from RNA encoded by said gene. 
     
     
         25 . The method of  claim 14 , wherein determining the level is performed by quantifying EST generated from RNA encoded by the gene. 
     
     
         26 . The method of  claim 14 , wherein the level of RNA encoded by the gene is determined relative to a level of RNA encoded by an internal control gene in the blood sample. 
     
     
         27 . A method of identifying a potential marker of a disease, the method comprising:
 (a) determining a level of RNA encoded by a gene in blood samples of human subjects having the disease, and a level of RNA encoded by the gene in blood samples of human subjects not having the disease, wherein the gene is KIAA0796; and   (b) comparing the levels with each other,   wherein a determination, resulting from step (b), of a significant difference between the levels identifies the gene as a potential marker of the disease.   
     
     
         28 . The method of  claim 27 , wherein determining the level in blood samples of human subjects having the disease, and/or wherein determining the level in blood samples of human subjects not having the disease is effected using at least one oligonucleotide of predetermined sequence. 
     
     
         29 . The method of  claim 28 , wherein the at least one oligonucleotide is specific for RNA encoded only by the gene in blood of human subjects, and/or is specific for cDNA complementary to RNA encoded only by the gene in blood of human subjects. 
     
     
         30 . The method of  claim 28 , wherein determining the level in blood samples of human subjects having the disease, and/or wherein determining the level in blood samples of human subjects not having the disease is effected by amplifying RNA encoded by the gene to form amplified product, using at least one primer, and quantifying the amplified product, wherein the at least one oligonucleotide comprises the at least one primer. 
     
     
         31 . The method of  claim 28 , wherein determining the level in blood samples of human subjects having the disease, and/or wherein determining the level in blood samples of human subjects not having the disease is effected by hybridizing cDNA complementary to RNA encoded by the gene with at least one immobilized probe to form hybridization product, and quantifying the hybridization product, wherein the at least one oligonucleotide comprises the at least one probe. 
     
     
         32 . The method of  claim 27 , wherein determining the level in blood samples of human subjects having the disease is effected by quantifying cDNA generated from RNA encoded by the gene and/or by quantifying EST generated from RNA encoded by the gene; and/or wherein determining the level in blood samples of human subjects not having the disease is effected by quantifying cDNA generated from RNA encoded by the gene and/or by quantifying EST generated from RNA encoded by the gene. 
     
     
         33 . The method of  claim 27 , wherein the level in the blood samples of subjects having the disease is relative to a level of RNA encoded by a housekeeping gene in the blood samples of subjects having the disease; and/or wherein the level in the blood samples of subjects not having the disease is relative to a level of RNA encoded by a housekeeping gene in the blood samples of subjects not having the disease. 
     
     
         34 . The method of  claim 27 , wherein the subjects not having the disease are healthy subjects. 
     
     
         35 . The method of  claim 27 , wherein the blood samples of human subjects having the disease comprise one or more cell types selected from the group consisting of: granulocytes, monocytes, CD19+ cells and CD3+ cells; and/or wherein the blood samples of human subjects not having the disease comprise one or more cell types selected from the group consisting of: granulocytes, monocytes, CD19+ cells and CD3+ cells. 
     
     
         36 . The method of  claim 27 , wherein the difference is a decrease in the level in the blood samples of subjects having the disease relative to the level in the blood samples of subjects not having the disease. 
     
     
         37 . A method of identifying a potential marker for differentiating between different body states, the method comprising:
 (a) determining a level of RNA encoded by a gene in blood samples of human subjects having a first body state, and a level of RNA encoded by the gene in blood samples of human subjects having a second body state, wherein the second body state is different from the first body state, and wherein the gene is KIAA0796; and   (b) comparing the levels with each other,   wherein a determination, resulting from step (b), of a significant difference between the levels identifies the gene as a potential marker for differentiating between the first body state and the second body state.   
     
     
         38 . The method of  claim 37 , wherein determining the level in blood samples of human subjects having the first body state, and/or wherein determining the level in blood samples of human subjects having the second body state is effected using at least one oligonucleotide of predetermined sequence. 
     
     
         39 . The method of  claim 38 , wherein the at least one oligonucleotide is specific for RNA encoded only by the gene in blood of human subjects, and/or is specific for cDNA complementary to RNA encoded only by the gene in blood of human subjects. 
     
     
         40 . The method of  claim 38 , wherein determining the level in blood samples of human subjects having the first body state, and/or wherein determining the level in blood samples of human subjects having the second body state is effected by amplifying RNA encoded by the gene to form amplified product, using at least one primer, and quantifying the amplified product, wherein the at least one oligonucleotide comprises the at least one primer. 
     
     
         41 . The method of  claim 37  wherein the level in the blood samples of subjects having the first body state is relative to a level of RNA encoded by a housekeeping gene in the blood samples of subjects having the first body state; and/or wherein the level in the blood samples of subjects having the second body state is relative to a level of RNA encoded by a housekeeping gene in the blood samples of subjects having the second body state. 
     
     
         42 . The method of  claim 38 , wherein determining the level in blood samples of human subjects having the first body state, and/or wherein determining the level in blood samples of human subjects having the second body state is effected by hybridizing cDNA complementary to RNA encoded by the gene with at least one immobilized probe to form hybridization product, and quantifying the hybridization product, wherein the at least one oligonucleotide comprises the at least one probe.