Method for the detection of cancer

Abstract

The present invention relates to a method for the diagnosis and/or the follow up of the evolution of cancer, which includes the analysis and quantification of over expressed and amplified genes in the plasma/serum of cancer patients or persons suspected to harbor cancer. This is achieved by analyzing together the amount of DNA and RNA of certain genes in the plasma/serum of cancer patients that are the reflection of a gene amplification and/or a gene over expression in comparison to healthy controls.

The invention claimed is: 
     
       1. A method for the diagnosis or the follow up of the evolution of cancers which comprises measuring both gene over-expression (RNA) and gene amplification (DNA) of a gene present in the a sample derived from bodily fluids of fluid from a patient, that is both amplified and over-expressed in cancer cells and comparing to healthy controls, wherein the sample is cell free or substantially cell free. 
     
     
       2. The method according to  claim 1 , wherein the RNA and the DNA are extracted from a the bodily fluid, purified and amplified, and the over-expressed RNA and the amplified DNA are analyzed and compared to a house keeping gene. 
     
     
       3. The method according to  claim 2 , wherein the genes gene analyzed are is selected from hTERT, hTR, TEP1, MYCN, MYCC, ErbB2, Her2, Her2/Neu, Her 1, Cyclin A and D1, ABL, SKP2, ETV6 (TELgene), MGC2177, PLAG1, PSMC6P and LYN. 
     
     
       4. The method according to  claim 2 , wherein the nucleic acids RNA and the DNA are amplified by reverse transcriptase chain reaction (RT-PCR). 
     
     
       5. The method according to  claim 3 , wherein the nucleic acids RNA and the DNA are amplified by reverse transcriptase chain reaction (RT-PCR). 
     
     
       6. The method according to  claim 1 , wherein the genes analyzed are gene measured is selected from the group consisting of hTERT, hTR, TEP1, MYCN, MYCC, ErbB2, Her2, Her2/Neu, Her 1, Cyclin A and D1, ABL, SKP2, ETV6 (TELgene), MGC2177, PLAG1, PSMC6P and LYN. 
     
     
       7. The method according to  claim 6 , wherein the nucleic acids RNA and the DNA are amplified by reverse transcriptase chain reaction (RT-PCR). 
     
     
       8. The method according to  claim 1 , wherein the nucleic acids RNA and the DNA are amplified by reverse transcriptase chain reaction (RT-PCR). 
     
     
       9. The method according to  claim 1 , wherein the genes analyzed are gene measured is compared to a reference nucleic acid extract (DNA and RNA) corresponding to the expression (RNA) and quantity (DNA) of a house keeping gene. 
     
     
       10. The method according to  claim 1 , wherein the genes analyzed are gene is compared to a reference RNA corresponding to the expression of a house keeping gene. 
     
     
       11. The method according to  claim 1 , wherein the genes analyzed are gene is compared to a reference DNA corresponding to a housekeeping gene. 
     
     
       12. The method according to  claim 1 , wherein the gene quantification may be estimated in reference to a standard curve obtained with nucleic acids of a cell line. 
     
     
       13. The method according to  claim 1 , wherein the nucleic acids RNA and the DNA are analyzed by gel coloration, by radioactive immunological technique (RIA), by enzyme linked immunosorbant test (ELISA) or by a microchip test (gene array), and quantified by any method for nucleic acid quantification. 
     
     
       14. The method according to  claim 1 , wherein the nucleic acids RNA and DNA are is quantified by real time RT PCR RT-PCR. 
     
     
       15. The method according to  claim 1 , wherein the RNA and the DNA are extracted from the patient and measured simultaneously. 
     
     
       16. A method for measuring both gene over-expression (RNA) and gene amplification (DNA) in a patient suspected of having cancer, wherein the gene measured is selected from the group consisting of hTERT, hTR, TEP1, MYCN, MYCC, ErbB2, Her2, Her2/Neu, Her 1, Cyclin A, Cyclin D1, ABL, SKP2, ETV6 (TELgene), MGC2177, PLAG1, PSMC6P and LYN, comprising: obtaining a sample of derived from a bodily fluid from a patient, and wherein the sample is cell free or substantially cell free, extracting both RNA and DNA simultaneously from said sample, measuring both gene over-expression (RNA) and gene amplification (DNA) at the same time in said sample from the extracted RNA and DNA, and comparing said measurement to a healthy control. 
     
     
       17. The method according to  claim 16 , wherein the RNA and the DNA are extracted from a the bodily fluid, purified and amplified, and the over-expressed RNA and the amplified DNA are analyzed and compared to a house keeping gene. 
     
     
       18. The method according to  claim 17 , wherein the over-expressed and amplified gene is hTERT. 
     
     
       19. The method according to  claim 18 , wherein the nucleic acids are RNA is amplified by reverse transcriptase polymerase chain reaction (RT-PCR). 
     
     
       20. The method according to  claim 16 , wherein the genes analyzed are gene measured is compared to a reference nucleic acid extract (DNA and RNA) corresponding to the expression (RNA) and quantity (DNA) of a house keeping gene. 
     
     
       21. The method according to  claim 16 , wherein the genes gene analyzed are is compared to a reference RNA corresponding to the expression of a house keeping gene. 
     
     
       22. The method according to claim 1, wherein said sample is selected from plasma, serum, saliva, and sputum. 
     
     
       23. The method according to claim 16, wherein said sample is selected from plasma, serum, saliva, and sputum. 
     
     
       24. The method according to either claim 1 or 16, wherein said RNA and said DNA from said bodily fluid comprise extracellular RNA and DNA. 
     
     
       25. The method according to either claim 1 or 16, wherein said measuring comprises amplifying both of said RNA and said DNA by polymerase chain reaction. 
     
     
       26. The method according to claim 25, wherein said amplifying further comprises using a single primer/probe set to amplify both of said DNA and said RNA. 
     
     
       27. A method for the diagnosis or the follow up of the evolution of cancers which comprises measuring both the quantity of gene expression (RNA) and quantity of gene amplification (DNA) of one or more genes in a sample derived from a bodily fluid of a patient, wherein said sample is cell free or substantially cell free, and comparing to healthy controls. 
     
     
       28. The method according to claim 27, wherein said measuring comprises extracting both of said RNA and said DNA from said bodily fluid, purifying said extracted RNA and said extracted DNA, amplifying said purified RNA and said purified DNA, analyzing said amplified RNA and said amplified DNA, and comparing to a house keeping gene. 
     
     
       29. The method according to claim 28, wherein said amplifying comprises using polymerase chain reaction. 
     
     
       30. The method according to claim 28, wherein said amplifying comprises using a single primer/probe set to amplify both of said RNA and said DNA. 
     
     
       31. The method according to claim 27, wherein said comparing comprises comparing to a reference nucleic acid extract corresponding to expression (RNA) and/or quantity (DNA) of a house keeping gene. 
     
     
       32. The method according to claim 27, wherein said measuring comprises estimating said one or more genes in reference to a standard curve obtained with nucleic acids of a cell line. 
     
     
       33. The method according to claim 27, further comprising analyzing said RNA and said DNA by gel coloration, by radioactive immunological technique (RIA), by enzyme linked immunosorbant test (ELISA) or by a microchip test (gene array), and quantifying by any method for nucleic acid quantification. 
     
     
       34. The method according to claim 27, further comprising extracting both of said RNA and said DNA from the patient and measuring simultaneously. 
     
     
       35. The method according to claim 27, wherein said sample is selected from plasma, serum, saliva, and sputum. 
     
     
       36. The method according to claim 27, wherein both of said RNA and said DNA comprise extracellular RNA and extracellular DNA. 
     
     
       37. The method according to claim 1 or 16 further comprising providing a diagnosis and/or follow up as to the evolution of said cancer. 
     
     
       38. The method of claim 1, wherein the measuring is performed using a microchip. 
     
     
       39. The method of claim 16, wherein the measuring is performed using a microchip. 
     
     
       40. The method of claim 27, wherein the measuring is performed using a microchip. 
     
     
       41. The method of claim 1, further comprising identifying if the patient is in need of treatment if there is an increased level in the sample as compared to the healthy controls. 
     
     
       42. The method of claim 16, further comprising identifying if the patient is in need of treatment if there is an increased level in the sample as compared to the healthy controls. 
     
     
       43. The method of claim 27, further comprising identifying if the patient is in need of treatment if there is an increased level in the sample as compared to the healthy controls. 
     
     
       44. The method of claim 41, further comprising providing treatment to the identified patient. 
     
     
       45. The method of claim 42, further comprising providing treatment to the identified patient. 
     
     
       46. The method of claim 43, further comprising providing treatment to the identified patient.