WO2007116417A1 - Nouveaux marqueurs de méthylation d'adn utilisés dans le diagnostic de maladies néoplastiques - Google Patents

Nouveaux marqueurs de méthylation d'adn utilisés dans le diagnostic de maladies néoplastiques Download PDF

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WO2007116417A1
WO2007116417A1 PCT/IT2006/000237 IT2006000237W WO2007116417A1 WO 2007116417 A1 WO2007116417 A1 WO 2007116417A1 IT 2006000237 W IT2006000237 W IT 2006000237W WO 2007116417 A1 WO2007116417 A1 WO 2007116417A1
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seq
sequence
tumours
methylation
nucleotides
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PCT/IT2006/000237
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Silvia Sabbioni
Elena Miotto
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Silvia Sabbioni
Elena Miotto
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Priority to PCT/IT2006/000237 priority Critical patent/WO2007116417A1/fr
Publication of WO2007116417A1 publication Critical patent/WO2007116417A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/154—Methylation markers

Definitions

  • Novel DNA methylation markers useful for the diagnosis of neoplastic diseases
  • the present invention relates to an in vitro method for the diagnosis of neoplastic diseases in humans through the analysis of the methylation state of the DNA of genes which are useful as tumour markers, as well as to nucleic acids and oligonucleotides useful in such method and to kits comprising such oligonucleotides.
  • DNA methylation is known to have a regulatory function, in particular of gene transcription. It has in fact been observed that the gene promoters often contain regions which are particularly rich in CpG dinucleotides, known as CpG islands, the cytosines of which are generally unmethylated when the gene is transcriptionally active.
  • neoplastic diseases are characterised by the altered functionality of specific genes involved in the normal physiology of the cell, and in the scientific literature several oncosuppressor genes which are activated by the abnormal methylation of their own promoter are described.
  • the inventors have found that the methylation status of the CpG islands of the genes listed in table 1 is altered in human tumours and that the analysis of the methylation status of the CpG islands of such genes is a useful marker for the early diagnosis of various types of human neoplastic diseases, comprising solid tumours and haematological neoplastic diseases.
  • the inventors have found that the CpG islands located at the 5' end of the genes listed in table 1 have an higher methylation rate in cells derived from human neoplastic diseases than in normal controls which are unmethylated or have a low methylation rate.
  • the CpG islands of the genes listed in table 1 are all located at the 5 1 end of such genes and they cover regions varying from about 2.6 kb to about 8.0 kb, which include the 5' region, the first exon and a portion of the first intron of each gene.
  • the present invention provides a method for the diagnosis of solid tumours and haematological neoplastic diseases, characterised in that it comprises, in a sample of genomic DNA, the step of analysing the methylation status of the CpG island of a gene selected from the group consisting of the human genes listed in table 1 , the genomic location of the CpG island of each of such genes being as shown in table 1, the methylation od said CpG island being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the ALDH13A gene comprised between the nucleotide positions corresponding to nucleotides 86.701 and 89.700 of the Genbank sequence AC015712, version 10 (SEQ ID NO: 1), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the ARHGAP20 gene comprised between the nucleotide positions corresponding to nucleotides 42.661 and 45.840 of the Genbank sequence AP003460, version 3 (SEQ ID NO: 2), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the FLI-1 gene comprised between the nucleotide positions corresponding to nucleotides 40.001 and 47.460 of the Genbank sequence AP001122, version 5 (SEQ ID NO: 3), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the FOSL2 gene comprised between the nucleotide positions corresponding to nucleotides 52.801 and 57.000 of the Genbank sequence AC104695, version 2 (SEQ ID NO: 4), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the HES2 gene comprised between the nucleotide positions corresponding to nucleotides 26.641 and 34.670 of the Genbank sequence AL031848, version 11 (SEQ ID NO: 5), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the MAPI B gene comprised between the nucleotide positions corresponding to nucleotides 38.041 and 42.000 of the Genbank sequence AC093218, version 2 (SEQ ID NO: 6), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the MDM2 gene comprised between the nucleotide positions corresponding to nucleotides 1.001 and 3.000 of the Genbank sequence AF527840, version 1 (SEQ ID NO: 7), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the NID1 gene comprised between the nucleotide positions corresponding to nucleotides 985.000 and 986.859 of the Genbank sequence NT_004836, version 16 (SEQ ID NO: 8), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the NID2 gene comprised between the nucleotide positions corresponding to nucleotides 33.534.100 and 33.536.379 of the Genbank sequence NT_026437, version 11 (SEQ ID NO: 9), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the PNUTL2 gene comprised between the nucleotide positions corresponding to nucleotides 108.541 and 111.200 of the Genbank sequence AC005666, version 1 (SEQ ID NO: 10), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the PTGER3 gene comprised between the nucleotide positions corresponding to nucleotides 122.263 and 126.262 of the Genbank sequence AL031429, version 11 (SEQ ID NO: 11), the methylation of said sequence being indicative of neoplastic disease.
  • the method of the invention comprises the step of analysing the methylation status of the nucleic acid sequence of the PTPRM gene comprised between the nucleotide positions corresponding to nucleotides 63.001 and 67.500 of the Genbank sequence AP001091 , version 5 (SEQ ID NO: 12), the methylation of said sequence being indicative of neoplastic disease.
  • Table 1 summarises the correspondence between the CpG islands of each of the above- mentioned genes and the nucleotide positions of the relevent Genbank sequences.
  • Table 1 Novel abnormal ene meth lation markers in human tumours and location of their C G islands
  • the methylation status of of the CpG island is analysed using techniques which include chemical treatment of the DNA with a reagent capable of converting methylation differences into sequence differences.
  • a chemical reagent is used to this end which is capable of converting the unmethylated cytosine bases into uracil or any other base capable of pairing with a base other than guanine.
  • One preferred reagent for this purpose is bisulfite, preferably sodium bisulfite, in combination with hydroquinone.
  • the unmethylated cytosines of DNA are in fact deaminated to uracil which, in terms of base pairing, corresponds to thymidine.
  • the methylated cytosines are resistant to the chemical treatment and thus do not undergo any modification. Consequently, the methylcytosines of the original genomic DNA, which could not initially be distinguished from the cytosines because they exhibited the same pairing characteristics, remain the only cytosines in the chemicallly-treated DNA and they may thus be identified using standard molecular biology techniques.
  • the base sequence of the chemicallly-treated DNA may be analysed for example by direct sequencing.
  • direct sequencing it is possible to use techniques such as Methylation- Specific PCR (MSP), which are based on the methylation-specific amplification of segments of the CpG island of the sample of the chemically-treated DNA.
  • MSP Methylation- Specific PCR
  • Such methods make use of oligonucleotides which are capable of undergoing selective hybridisation with the modified sequences of the methylated or unmethylated CpG island obtained with the above-described chemical treatment.
  • sequences designated as SEQ ID NO: 13 and SEQ ID NO: 14 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the ALDH1A3 gene are methylated.
  • sequences designated as SEQ ID NO: 15 and SEQ ID NO: 16 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the ALDH 1 A3 gene is methylated.
  • sequences designated as SEQ ID NO: 17 and SEQ ID NO: 18 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the ARHGAP20 gene are methylated.
  • sequences designated as SEQ ID NO: 19 and SEQ ID NO: 20 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the ARHGAP20 gene is methylated.
  • sequences designated as SEQ ID NO: 21 and SEQ ID NO: 22 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the FLU gene are methylated.
  • sequences designated as SEQ ID NO: 23 and SEQ ID NO: 24 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the FLU gene is methylated.
  • sequences designated as SEQ ID NO: 25 and SEQ ID NO: 26 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the FOSL2 gene are methylated.
  • sequences designated as SEQ ID NO: 27 and SEQ ID NO: 28 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the FOSL2 gene is methylated.
  • sequences designated as SEQ ID NO: 29 and SEQ ID NO: 30 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the HES2 gene are methylated.
  • sequences designated as SEQ ID NO: 31 and SEQ ID NO: 32 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the HES2 gene is methylated.
  • sequences designated as SEQ ID NO: 33 and SEQ ID NO: 34 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the MAPI B gene are methylated.
  • sequences designated as SEQ ID NO: 35 and SEQ ID NO: 36 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the MAPI B gene is methylated.
  • sequences designated as SEQ ID NO: 37 and SEQ ID NO: 38 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the MDM2 gene are methylated.
  • sequences designated as SEQ ID NO: 39 and SEQ ID NO: 40 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the MDM2 gene is methylated.
  • sequences designated as SEQ ID NO: 41 and SEQ ID NO: 42 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the NID1 gene are methylated.
  • sequences designated as SEQ ID NO: 43 and SEQ ID NO: 44 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the NID1 gene is methylated.
  • sequences designated as SEQ ID NO: 45 and SEQ ID NO: 46 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the NID2 gene are methylated.
  • sequences designated as SEQ ID NO: 47 and SEQ ID NO: 48 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the NID2 gene is methylated.
  • sequences designated as SEQ ID NO: 49 and SEQ ID NO: 50 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the PNUTL2 gene are methylated.
  • sequences designated as SEQ ID NO: 51 and SEQ ID NO: 52 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the PNUTL2 gene is methylated.
  • sequences designated as SEQ ID NO: 53 and SEQ ID NO: 54 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the PTGER3 gene are methylated.
  • sequences designated as SEQ ID NO: 55 and SEQ ID NO: 56 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the PTGER3 gene is methylated.
  • sequences designated as SEQ ID NO: 57 and SEQ ID NO: 58 are the nucleic acid sequences of the two modified strands which are obtained when all the dinucleotides of the original CpG island of the PTPRM gene are methylated.
  • sequences designated as SEQ ID NO: 59 and SEQ ID NO: 60 are the nucleic acid sequences of the two modified strands which are obtained when none of the dinucleotides of the original CpG island of the PTPRM gene is methylated.
  • Sequences SEQ ID NO: 13 , SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16 were determined on the basis of Genbank sequence AC015712, version 10 (nucleotides 86701-89700) of the ALDH 1 A3 gene.
  • Sequences SEQ ID NO: 17 , SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20 were determined on the basis of Genbank sequence AP003460, version 3 (nucleotides 42661- 45840) of the ARHGAP20 gene.
  • Sequences SEQ ID NO: 21 , SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24 were determined on the basis of Genbank sequence AP001122, version 5 (nucleotides 40001- 46400) of the FLU gene.
  • Sequences SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27 and SEQ ID NO: 28 were determined on the basis of Genbank sequence AC104695, version 2 (nucleotides 52801- 57000) of the FOSL2 gene. Sequences SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32 were determined on the basis of Genbank sequence AL031848, version 11 (nucleotides 26641-34670) of the HES2 gene.
  • Sequences SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36 were determined on the basis of Genbank sequence AC093218, version 2 (nucleotides 38041- 42000) of the MAPI B gene.
  • Sequences SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39 and SEQ ID NO: 40 were determined on the basis- of Genbank sequence AF527840, version 1 (nucleotides 1.001- 3.000) of the MDM2 gene.
  • Sequences SEQ ID NO: 41 , SEQ ID NO: 42, SEQ ID NO: 43 and SEQ ID NO: 44 were determined on the basis of Genbank sequence NT_004836, version 16 (nucleotides 985.000 - 986.859) of the NID1 gene.
  • Sequences SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48 were determined on the basis of Genbank sequence NT_026437, version 11 (nucleotides 33.534.100 - 33.536.379) of the NID2 gene.
  • Sequences SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 and SEQ ID NO: 52 were determined on the basis of Genbank sequence AC005666, version 1 (nucleotides 108541-111200) of the PNUTL2 gene.
  • Sequences SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56 were determined on the basis of Genbank sequence AL031429, version 11 (nucleotides 122263-126262) of the PTGER3 gene.
  • Sequences SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59 and SEQ ID NO: 60 were determined on the basis of Genbank sequence AP001091, version 5 (nucleotides 63001- 67500) of the PTPRM gene.
  • Oligonucleotides at least 10 nucleotides in length which are complementary or identical to a segment of a target sequence selected from the group consisting of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21 , SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31 , SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 , SEQ ID NO: 42, SEQ ID NO:
  • oligonucleotides fall within the scope of the present invention.
  • the oligonucleotides of the invention are complementary or identical to a segment of a target sequence as defined above comprising at least one CpG dinucleotide.
  • oligonucleotides may be used in the diagnostic method of the invention as amplification primers or as hybridisation probes. In the latter case, they preferably have a length of at least 18 nucleotides.
  • table 2a reports some specific oligonucleotide primers based on some of the target sequences listed above, which are useful in the diagnostic method of the invention.
  • the diagnostic method of the invention may, for example, be a detection method based on DNA amplification, optionally followed by detection of the amplified DNA fragments by a hybridisation probe.
  • a sample of genomic DNA to be investigated is chemically pretreated with a reagent capable of converting the unmethylated cytosine bases into uracil or any other base capable of pairing with a base other than guanine.
  • the sample of genomic DNA to be analysed is preferably obtained from a source such as blood, serum, plasma, bronchial washings, expectoration, saliva, intestinal washings, urine, faeces, ejaculate, pads of various origins, needle aspirates, lymph node biopsies, or combinations thereof.
  • a source such as blood, serum, plasma, bronchial washings, expectoration, saliva, intestinal washings, urine, faeces, ejaculate, pads of various origins, needle aspirates, lymph node biopsies, or combinations thereof.
  • Said chemical pretreatment of the genomic DNA is preferably carried out with sodium bisulfite and hydroquinone.
  • any of the modified strands obtained by pretreatment of the genomic DNA may be subjected to an amplification reaction using as the amplification primers a pair of oligonucleotides at least 10 nucleotides in length, wherein one of said oligonucleotides is complementary to a first segment of a target sequence selected from the group consisting of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 57 and SEQ ID NO: 58
  • At least one of said first and second segments of the target sequence comprises at least one CpG dinucleotide.
  • the amplification reaction is methylation-specific, since the oligonucleotide primer pair used is designated to specifically amplify a modified strand derived from the sequence of the original methylated CpG island.
  • detection of the amplified DNA fragments may be performed by any method known per se, for example by electrophoresis on agarose gel and staining with a DNA intercalating agent, such as for example ethidium bromide.
  • neither the first nor the second segment of the target sequence comprises a CpG dinucleotide.
  • the amplification reaction is not methylation-specific. Consequently, the detection of the amplified fragments, if present, will be performed by hybridisation with an oligonucleotide probe at (east 18 nucleotides in length, which is complementary to a third segment of the target sequence, said third segment being located between the said first and second segments and comprising at least one CpG dinucleotide.
  • the third segment of the target sequence may optionally be partially overlapped to the first or the second segment.
  • the amplification reaction is preferably a polymerase chain reaction (PCR). Still more preferred is a two-step amplification reaction comprising a primary PCR followed by a semi-nested or nested secondary PCR.
  • PCR polymerase chain reaction
  • the modified genomic DNA representing the target of the amplification reaction is initially single-stranded
  • only one of the two primers will initially hybridise with the target strand.
  • the first hybridised primer will be extended by a polymerase using the target strand as template.
  • the second primer will hybridise with the newly synthesised chain and will be extended by the polymerase using the newly synthesised chain as the template.
  • the initial single-stranded target DNA will be converted into double-stranded DNA.
  • the two amplification primers will each simultaneously hybridise with one strand of the double-stranded target DNA.
  • Table 2a shows some specific examples of oligonucleotides suitable as primers or probes in the diagnostic method of the invention.
  • the oligonucleotides ALDH1A3_MF (SEQ ID NO: 61) and ALDH1A3_MR (SEQ ID NO: 62) were designed to specifically hybridise in SEQ ID NO: 13.
  • the oligonucleotide pair ALDH1A3_MF and ALDH1A3_MR amplifies a 227 bp sequence comprised between nucleotide 1238 and nucleotide 1465 of SEQ ID NO: 13.
  • the oligonucleotides ARHGAP20_MF (SEQ ID NO: 63) and ARHGAP20_MR (SEQ ID NO: 64) were designed to specifically hybridise in SEQ ID NO: 18.
  • the oligonucleotide pair ARHGAP20_MF and ARHGAP20_MR amplifies a 153 bp sequence comprised between nucleotide 1777 and nucleotide 1930 of SEQ ID NO: 18.
  • the oligonucleotides FLI1_MF (SEQ ID NO: 65) and FLI1_MR (SEQ ID NO: 66) were designed to specifically hybridise in SEQ ID NO: 21.
  • the oligonucleotide pair FLI1_MF and FL11_MR amplifies a 199 bp sequence comprised between nucleotide 5744 and nucleotide 5943 of SEQ ID NO: 21.
  • the oligonucleotides FOSL2_MF (SEQ ID NO: 67) and FOSL2_MR (SEQ ID NO: 68) were designed to specifically hybridise in SEQ ID NO: 25.
  • the oligonucleotide pair FOSL2_MF and FOSL2_MR amplifies a 264 bp sequence comprised between nucleotide 2496 and nucleotide 2760 of SEQ ID NO: 25.
  • the oligonucleotides HES2_MF (SEQ ID NO: 69) and HES2_MR (SEQ ID NO: 70) were designed to specifically hybridise in SEQ ID NO: 29.
  • the oligonucleotide pair HES2_MF and HES2__MR amplifies a 205 bp sequence comprised between nucleotide 6072 and nucleotide 6277 of SEQ ID NO: 29
  • oligonucleotides MAP1B_MF SEQ ID NO: 71
  • MAP1B_MR SEQ ID NO: 72
  • MAP1B_MF and MAP1B_MR amplifies a 216 bp sequence comprised between nucleotide 1635 and nucleotide 1851 of SEQ ID NO: 33.
  • the oligonucleotides MDM2_MF (SEQ ID NO: 73) and MDM2_MR (SEQ ID NO: 74) were designed to specifically hybridise in SEQ ID NO: 37.
  • the oligonucleotide pair MDM2_MF and MDM2_MR amplifies a 328 bp sequence comprised between nucleotide 1175 and nucleotide 1503 of SEQ ID NO: 37.
  • the oligonucleotides NID1_MF (SEQ ID NO: 75) and NID1_MR (SEQ ID NO: 76) were designed to specifically hybridise in SEQ ID NO: 41.
  • the oligonucleotide pair NID1_MF and NID1_MR amplifies a 144 bp sequence comprised between nucleotide 1319 and nucleotide 1462 of SEQ ID NO: 41.
  • the oligonucleotides NID2_MF (SEQ ID NO: 77) and NID2_MR (SEQ ID NO: 78) were designed to specifically hybridise in SEQ ID NO: 45.
  • the oligonucleotide pair NID2_MF and NID2_MR amplifies a 141 bp sequence comprised between nucleotide 1310 and nucleotide 1438 of SEQ ID NO: 45
  • the oligonucleotides PNUTL2_MF (SEQ ID NO: 79) and PNUTL2_MR (SEQ ID NO: 80) were designed to specifically hybridise in SEQ ID NO: 49.
  • the oligonucleotide pair PNUTL2_MF and PNUTL2_MR amplifies a 216 bp sequence comprised between nucleotide 826 and nucleotide 1042 of SEQ ID NO: 49.
  • oligonucleotides PTGER3_MF SEQ ID NO: 81
  • PTGER3_MR SEQ ID NO: 82
  • oligonucleotide pair PTGER3_MF and PTGER3_MR amplifies a 191 bp sequence comprised between nucleotide 2089 and nucleotide 2280 of SEQ ID NO: 54.
  • the oligonucleotides PTPRM_MF (SEQ ID NO: 83) and PTPRM_MR (SEQ ID NO: 84) were designed to specifically hybridise in SEQ ID NO: 57.
  • the oligonucleotide pair PTPRM-MF and PTPRM_MR amplifies a 245 bp sequence comprised between nucleotide 2061 and nucleotide 2306 of SEQ ID NO: 57.
  • kit for performing the diagnostic method of the invention also falls within the scope of the present invention, which kit may comprise, for example, a pair of amplification primers as defined above optionally in combination with a reagent capable of converting the unmethylated cytosine bases into uracil or any other base capable of pairing with a base other than guanine, for example sodium bisulfite in combination with hydroquinone.
  • the kit may further comprise DNA detection means, such as for example a methylation- specific hybridisation probe as defined above, and/or DNA amplification means, for example a polymerase enzyme.
  • the kit may comprise a DNA detection reagent such as a DNA intercalating agent, for example ethidium bromide.
  • the kit of the invention may also comprise a second pair of amplification primers at least 10 nucleotides in length, one of said oligonucleotides being complementary to a first segment of a target sequence selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 31 , SEQ ID NO: 32, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 51 , SEQ ID NO: 52, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 59, SEQ ID NO: 60 and the other oligonucleotide being identical to a second segment of said target sequence, the second segment being located downstream from the first segment
  • At least one of said first and second segments of the target sequence comprises at least one TpG dinucleotide.
  • the second pair of primers is used as a control that the DNA actually has been modified following treatment with bisulfite, so making it possible to validate a negative result of amplification with primers designed on the basis of the modified nucleotide sequences derived from the methylated CpG island.
  • Colon Mucosa 0% (0/10) 0% (0/10) 21% . (5/24) 70% (7/10) 0% (0/10) 40% (4/10) 33% (8/24)
  • Liver Mucosa 0% (0/9) 0% (0/9) 22% (2/9) 22% (2/9) 0% (0/9) 0% (0/9) 0% (0/9) 0% (0/9)
  • the methylation percentages of each gene in the various tissues analysed may be calculated.
  • the analysed tissues derive from colon, stomach, liver and breast. These results indicate that the genes listed in table 1 are often methylated in at least one of the tumour tissues analyzed. These tumours are to be considered as an example indicative of the high methylation frequency of such genes in neoplastic diseases, without limitation to the examples described in table 3.
  • SEQ ID NO. 1 is the CpG Island (isola CpG) of ALDH1A3, corresponding to the 3.000 nucleotide (nt) segment comprised between nt 86701-89700 of the Genbank sequence AC015712, versione 10; SEQ ID NO.
  • SEQ ID NO: 2 is the CpG island of ARHGAP20, corresponding to the 3.180 nt segment comprised between nt 45840 and 42661 of the Genbank sequence AP003460, version 3;
  • SEQ ID NO: 3 is the CpG Island of FLU corresponding to the 7.460 nt segment comprised between nt 40001-47460 of the Genbank sequence AP001122, version 5;
  • SEQ ID NO: 4 is the CpG Island of FOSL2 corresponding to the 4.200 nt segment comprised between nt 52801-57000 of the Genbank sequence AC104695, version 2;
  • SEQ ID NO: 5 is the CpG Island of HES2 corresponding to the 8.030 nt segment comprised between nt 26641- 34670 of the Genbank sequence AL031848, version 11;
  • SEQ ID NO: 6 is the CpG Island of MAPI B corresponding to the 3.960 nt segment comprised between nt 38041-42000 of the Genbank sequence
  • SEQ ID NO. 8 is the CpG island of the NID1 gene, corresponding to the 1.860 nt segment comprised between nt 985.000-986.859 of the Genbank sequence NT_004836, version 16;
  • SEQ ID NO. 9 is the CpG island of the NID2 gene, corresponding to the 2.280 nt segment comprised between nt 33534100- 33536379 of Genbank sequence NT_026437, version 11;
  • SEQ ID NO. 10 is the CpG island of the PNUTL2 gene, corresponding to the 2.660 nt segment comprised between nt 111200-108541 of Genbank AC005666, version 1;
  • SEQ ID NO. 11 is the CpG island of the PTGER3 gene, corresponding to the 4.000 nt segment comprised between nt 122.263 and 126.262 of Genbank AL031429, version 11;
  • SEQ ID NO. 12 is the CpG island of the PTPRM gene, corresponding to the 4.500 nt segment comprised between nt 63001 and 67500 of Genbank AP001091 , version 5;
  • SEQ ID NO. 13 is the sequence derived from the methylated sequence of the ALDH1A3 gene (top strand), derived from SEQ ID NO. 1;
  • SEQ ID NO. 14 is the sequence derived from the methylated sequence of the ALDH 1 A3 gene (bottom strand), derived from SEQ ID NO. 1;
  • SEQ ID NO. 15 is the sequence derived from the unmethylated sequence of the ALDH1A3 gene (top strand), derived from SEQ ID NO. 1;
  • SEQ ID NO. 16 is the sequence derived from the unmethylated sequence of the ALDH 1 A3 gene (bottom strand), derived from SEQ ID NO. 1;
  • SEQ ID NO. 17 is the sequence derived from the methylated sequence of the ARHGAP20 gene (top strand), derived from SEQ ID NO. 2;
  • SEQ ID NO. 18 is the sequence derived from the methylated sequence of the ARHGAP20 gene (bottom strand), derived from SEQ ID NO. 2;
  • SEQ ID NO. 19 is the sequence derived from the unmethylated sequence of the ARHGAP20 gene (top strand), derived from SEQ ID NO.
  • SEQ ID NO. 20 is the sequence derived from the unmethylated sequence of the ARHGAP20 gene (bottom strand), derived from SEQ ID NO. 2;
  • SEQ ID NO. 21 is the sequence derived from the methylated sequence of the FLU gene (top strand), derived from SEQ ID NO. 3;
  • SEQ ID NO. 22 is the sequence derived from the methylated sequence of the FLU gene (bottom strand), derived from SEQ ID NO. 3;
  • SEQ ID NO. 23 is the sequence derived from the unmethylated sequence of the FLU gene (top strand), derived from SEQ ID NO. 3;
  • SEQ ID NO. 24 is the sequence derived from the unmethylated sequence of the FLU gene (bottom strand), derived from SEQ ID NO.
  • SEQ ID NO: 25 is the sequence derived from the methylated sequence of the FOSL2 gene (top strand), derived from SEQ ID NO. 4;
  • SEQ ID NO: 26 is the sequence derived from the methylated sequence of the FOSL2 gene (bottom strand), derived from SEQ ID NO. 4;
  • SEQ ID NO: 27 is the sequence derived from the unmethylated sequence of the FOSL2 gene (top strand), derived from SEQ ID NO. 4;
  • SEQ ID NO: 28 is the sequence derived from the unmethylated sequence of the FOSL2 gene (bottom strand), derived from SEQ ID NO.
  • SEQ ID NO: 29 is the sequence derived from the methylated sequence of the HES2 gene (top strand), derived from SEQ ID NO.5;
  • SEQ ID NO: 30 is the sequence derived from the methylated sequence of the HES2 gene (bottom strand), derived from SEQ ID NO.5;
  • SEQ ID NO: 31 is the sequence derived from the unmethylated sequence of the HES2 gene (top strand), derived from SEQ ID NO.5;
  • SEQ ID NO: 32 is the sequence derived from the unmethylated sequence of the HES2 gene (bottom strand), derived from SEQ ID NO.5;
  • SEQ ID NO: 33 is the sequence derived from the methylated sequence of the MAPI B gene (top strand), derived from SEQ ID NO.6;
  • SEQ ID NO: 34 is the sequence derived from the methylated sequence of the MAPI B gene (bottom strand), derived from SEQ ID NO.6;
  • SEQ ID NO: 35 is the sequence derived from the un
  • SEQ ID NO: 39 is the sequence derived from the unmethylated sequence of the MDM2 gene (top strand), derived from SEQ ID NOJ;
  • SEQ ID NO: 40 is the sequence derived from the unmethylated sequence of the MDM2 gene (bottom strand), derived from SEQ ID NOJ;
  • SEQ ID NO: 41 is the sequence derived from the methylated sequence of the NID1 gene (top strand), derived from SEQ ID N0.8;
  • SEQ ID NO: 42 is the sequence derived from the methylated sequence of the NID1 gene (bottom strand), derived from SEQ ID N0.8;
  • SEQ ID NO: 41 is the sequence derived from the methylated sequence of the NID1 gene (top strand), derived from SEQ ID N0.8;
  • SEQ ID NO: 42 is the sequence derived from the methylated sequence of the NID1 gene (bottom strand), derived from SEQ ID N0.8;
  • SEQ ID NO: 44 is the sequence derived from the unmethylated sequence of the NID1 (bottom strand), derived from SEQ ID N0.8;
  • SEQ ID NO: 45 is the sequence derived from the methylated sequence of the NID2 gene (top strand), derived from SEQ ID N0.9;
  • SEQ ID NO: 46 is the sequence derived from the methylated sequence of the NID2 gene (bottom strand), derived from SEQ ID N0.9;
  • SEQ ID NO: 47 is the sequence derived from the unmethylated sequence of the NID2 gene (top strand), derived from SEQ ID N0.9;
  • SEQ ID NO: 48 is the sequence derived from the unmethylated sequence of the NID2 gene (bottom strand), derived from SEQ ID N0.9;
  • SEQ ID NO: 49 is the sequence derived from the methylated sequence of the PNUTL2 gene (top strand), derived from SEQ ID NO.10;
  • SEQ ID NO: 50 is the sequence derived from the methylated sequence of the PNUTL2 gene (bottom strand), derived from SEQ ID NO.10;
  • SEQ ID NO: 51 is the sequence derived from the unmethylated
  • SEQ ID NO: 62 is the oligonucleotide sequence for Methylation-Specific PCR (MSP) designed on the inverted complement of SEQ ID NO: 13;
  • SEQ ID NO: 63 is the oligonucleotide sequence for Methylation-Specific PCR (MSP) designed on SEQ ID NO: 18;
  • SEQ ID NO: 64 is the oligonucleotide sequence for Methylation-Specific PCR (MSP) designed on the inverted complement of SEQ ID NO: 18;
  • SEQ ID NO: 65 is the oligonucleotide sequence for Methylation-Specific PCR (MSP) designed on SEQ ID NO:21;
  • SEQ ID NO: 66 is the oligonucleotide sequence for Methylation-Specific PCR (MSP) designed on the inverted complement of SEQ ID NO:21;
  • SEQ ID NO: 67 is the oligonucleotide sequence for Methylation-Specific

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Abstract

L'invention concerne de nouveaux marqueurs de maladies néoplasiques humaines, ainsi que des méthodes, des oligonucléotides et des trousses pour les détecter. Lesdits marqueurs sont utilisés dans le diagnostic de tumeurs solides et de maladies hématologiques néoplasiques, sur base de l'analyse de l'état de méthylation d'îlots CpG d'un gène humain sélectionné parmi le groupe comportant ALDH 1 A3, ARHGAP20, FLU , FOSL2, HES2, MAPI B, MDM2, NID1 , NID2, PNUTL2, PTGER3 et PTPRM.
PCT/IT2006/000237 2006-04-07 2006-04-07 Nouveaux marqueurs de méthylation d'adn utilisés dans le diagnostic de maladies néoplastiques WO2007116417A1 (fr)

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Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009036922A2 (fr) * 2007-09-17 2009-03-26 Oncomethylome Sciences Sa Nouveaux marqueurs pour la détection du cancer de la vessie
WO2009115615A2 (fr) * 2008-03-21 2009-09-24 Oncomethylome Sciences S.A. Détection et pronostic du cancer du col de l’utérus
US20130224738A1 (en) * 2012-02-23 2013-08-29 Medical Diagnostic Laboratories, Llc Detecting dna methylation of bcl2, cdkn2a and nid2 genes to predict bladder cancer in humans
WO2015153283A1 (fr) * 2014-03-31 2015-10-08 Mayo Foundation For Medical Education And Research Détection de néoplasme colorectal
US9506116B2 (en) 2013-03-14 2016-11-29 Mayo Foundation For Medical Education And Research Detecting neoplasm
US10006093B2 (en) 2015-08-31 2018-06-26 Mayo Foundation For Medical Education And Research Detecting gastric neoplasm
US10030272B2 (en) 2015-02-27 2018-07-24 Mayo Foundation For Medical Education And Research Detecting gastrointestinal neoplasms
US10184154B2 (en) 2014-09-26 2019-01-22 Mayo Foundation For Medical Education And Research Detecting cholangiocarcinoma
US10196698B2 (en) 2013-01-23 2019-02-05 The Johns Hopkins University DNA methylation markers for metastatic prostate cancer
US10370726B2 (en) 2016-04-14 2019-08-06 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasia
US10435755B2 (en) 2015-03-27 2019-10-08 Exact Sciences Development Company, Llc Detecting esophageal disorders
US10435753B2 (en) 2010-03-26 2019-10-08 Mayo Foundation For Medical Education And Research Methods for detecting colorectal cancer using a DNA marker of exfoliated epithelia and a fecal blood marker
CN112063717A (zh) * 2020-09-17 2020-12-11 山东大学深圳研究院 Mdm2作为标志物在乙型肝炎病毒相关肝细胞癌早期诊断中的应用及检测试剂盒
US10934594B2 (en) 2017-11-30 2021-03-02 Mayo Foundation For Medical Education And Research Detecting breast cancer
US10934592B2 (en) 2017-02-28 2021-03-02 Mayo Foundation For Medical Education And Research Detecting prostate cancer
US11078543B2 (en) 2016-04-14 2021-08-03 Mayo Foundation For Medical Education And Research Detecting pancreatic high-grade dysplasia
CN113637754A (zh) * 2021-08-17 2021-11-12 武汉艾米森生命科技有限公司 生物标志物在诊断食管癌中的应用
CN114480661A (zh) * 2022-04-18 2022-05-13 北京起源聚禾生物科技有限公司 子宫内膜良恶性病变联合标志物、检测引物探针组及试剂盒

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002081749A2 (fr) * 2000-10-27 2002-10-17 University Of Southern California Sequences d'adn a methylation modifiee utilisees comme marqueurs associes au cancer humain
WO2003076593A2 (fr) * 2002-03-07 2003-09-18 The Johns Hopkins University School Of Medicine Depistage genomique pour genes lies au cancer rendus epigenetiquement silencieux
US20040072156A1 (en) * 2000-12-27 2004-04-15 Riken Institute Of Physical And Chemical Research Detection of genetic polymorphisms
US20050250137A1 (en) * 2002-09-20 2005-11-10 Tainsky Michael A Molecular targets of cancer and aging

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002081749A2 (fr) * 2000-10-27 2002-10-17 University Of Southern California Sequences d'adn a methylation modifiee utilisees comme marqueurs associes au cancer humain
US20040072156A1 (en) * 2000-12-27 2004-04-15 Riken Institute Of Physical And Chemical Research Detection of genetic polymorphisms
WO2003076593A2 (fr) * 2002-03-07 2003-09-18 The Johns Hopkins University School Of Medicine Depistage genomique pour genes lies au cancer rendus epigenetiquement silencieux
US20050250137A1 (en) * 2002-09-20 2005-11-10 Tainsky Michael A Molecular targets of cancer and aging

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
DATABASE EMBL 14 May 1999 (1999-05-14), XP002395926, Database accession no. U63607 *
DATABASE EMBL 17 December 1993 (1993-12-17), XP002395925, Database accession no. S64864 *
DATABASE EMBL 24 February 2000 (2000-02-24), XP002395924, Database accession no. AJ241944 *
DATABASE EMBL 29 September 2005 (2005-09-29), XP002395929, Database accession no. S44669 *
DATABASE EMBL 30 March 2005 (2005-03-30), XP002395928, Database accession no. AY845709 *
DATABASE EMBL 6 July 1989 (1989-07-06), XP002395930, Database accession no. V01083 *
DATABASE EMBL 7 May 1993 (1993-05-07), XP002395927, Database accession no. S85476 *
YAMASHITA SATOSHI ET AL: "Chemical genomic screening for methylation-silenced genes in gastric cancer cell lines using 5-aza-2 '-deoxycytidine treatment and oligonucleotide microarray", CANCER SCIENCE, vol. 97, no. 1, January 2006 (2006-01-01), pages 64 - 71, XP002395915, ISSN: 1347-9032 *

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WO2009036922A3 (fr) * 2007-09-17 2009-05-28 Oncomethylome Sciences Sa Nouveaux marqueurs pour la détection du cancer de la vessie
US9322065B2 (en) 2007-09-17 2016-04-26 Mdxhealth Sa Methods for determining methylation of the TWIST1 gene for bladder cancer detection
US10113202B2 (en) 2007-09-17 2018-10-30 Mdxhealth Sa Method for determining the methylation status of the promoter region of the TWIST1 gene in genomic DNA from bladder cells
WO2009036922A2 (fr) * 2007-09-17 2009-03-26 Oncomethylome Sciences Sa Nouveaux marqueurs pour la détection du cancer de la vessie
US10041128B2 (en) 2008-03-21 2018-08-07 MD×Health SA Methylation of the EPB41L3 gene or the promoter of the EPB41L3 gene in a test sample comprising cervical cells
WO2009115615A2 (fr) * 2008-03-21 2009-09-24 Oncomethylome Sciences S.A. Détection et pronostic du cancer du col de l’utérus
WO2009115615A3 (fr) * 2008-03-21 2009-11-12 Oncomethylome Sciences S.A. Détection et pronostic du cancer du col de l’utérus
US10435753B2 (en) 2010-03-26 2019-10-08 Mayo Foundation For Medical Education And Research Methods for detecting colorectal cancer using a DNA marker of exfoliated epithelia and a fecal blood marker
US20130224738A1 (en) * 2012-02-23 2013-08-29 Medical Diagnostic Laboratories, Llc Detecting dna methylation of bcl2, cdkn2a and nid2 genes to predict bladder cancer in humans
US9096905B2 (en) * 2012-02-23 2015-08-04 Medical Diagnostic Laboratories, Llc Detecting DNA methylation of BCL2, CDKN2A and NID2 genes to predict bladder cancer in humans
US10196698B2 (en) 2013-01-23 2019-02-05 The Johns Hopkins University DNA methylation markers for metastatic prostate cancer
US9994911B2 (en) 2013-03-14 2018-06-12 Mayo Foundation For Medical Education And Research Detecting neoplasm
US10683555B2 (en) 2013-03-14 2020-06-16 Mayo Foundation For Medical Education And Research Detecting neoplasm
US9982310B2 (en) 2013-03-14 2018-05-29 Mayo Foundation For Medical Education And Research Detecting neoplasm
US9506116B2 (en) 2013-03-14 2016-11-29 Mayo Foundation For Medical Education And Research Detecting neoplasm
US11821039B2 (en) 2013-03-14 2023-11-21 Mayo Foundation For Medical Education And Research Detecting neoplasm
US11987847B2 (en) 2014-03-31 2024-05-21 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasm
US11078539B2 (en) 2014-03-31 2021-08-03 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasm
US10301680B2 (en) 2014-03-31 2019-05-28 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasm
US10883144B2 (en) 2014-03-31 2021-01-05 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasm
WO2015153283A1 (fr) * 2014-03-31 2015-10-08 Mayo Foundation For Medical Education And Research Détection de néoplasme colorectal
US11365451B2 (en) 2014-03-31 2022-06-21 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasm
US10184154B2 (en) 2014-09-26 2019-01-22 Mayo Foundation For Medical Education And Research Detecting cholangiocarcinoma
US10900090B2 (en) 2014-09-26 2021-01-26 Mayo Foundation For Medical Education And Research Detecting cholangiocarcinoma
US10704107B2 (en) 2015-02-27 2020-07-07 Mayo Foundation For Medical Education And Research Detecting gastrointestinal neoplasms
US11384401B2 (en) 2015-02-27 2022-07-12 Mayo Foundation For Medical Education And Research Detecting gastrointestinal neoplasms
US10030272B2 (en) 2015-02-27 2018-07-24 Mayo Foundation For Medical Education And Research Detecting gastrointestinal neoplasms
US10435755B2 (en) 2015-03-27 2019-10-08 Exact Sciences Development Company, Llc Detecting esophageal disorders
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US10597733B2 (en) 2015-08-31 2020-03-24 Mayo Foundation For Medical Education And Research Detecting gastric neoplasm
US11542557B2 (en) 2016-04-14 2023-01-03 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasia
US11078543B2 (en) 2016-04-14 2021-08-03 Mayo Foundation For Medical Education And Research Detecting pancreatic high-grade dysplasia
US10370726B2 (en) 2016-04-14 2019-08-06 Mayo Foundation For Medical Education And Research Detecting colorectal neoplasia
US10934592B2 (en) 2017-02-28 2021-03-02 Mayo Foundation For Medical Education And Research Detecting prostate cancer
US11697853B2 (en) 2017-02-28 2023-07-11 Mayo Foundation For Medical Education And Research Detecting prostate cancer
US10934594B2 (en) 2017-11-30 2021-03-02 Mayo Foundation For Medical Education And Research Detecting breast cancer
US10975443B2 (en) 2017-11-30 2021-04-13 Mayo Foundation For Medical Education And Research Detecting breast cancer
CN112063717A (zh) * 2020-09-17 2020-12-11 山东大学深圳研究院 Mdm2作为标志物在乙型肝炎病毒相关肝细胞癌早期诊断中的应用及检测试剂盒
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