WO2022188776A1 - Marqueur de méthylation génique ou combinaison associée pouvant être utilisés pour le diagnostic compagnon du carcinome gastrique her2, et son utilisation - Google Patents

Marqueur de méthylation génique ou combinaison associée pouvant être utilisés pour le diagnostic compagnon du carcinome gastrique her2, et son utilisation Download PDF

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WO2022188776A1
WO2022188776A1 PCT/CN2022/079736 CN2022079736W WO2022188776A1 WO 2022188776 A1 WO2022188776 A1 WO 2022188776A1 CN 2022079736 W CN2022079736 W CN 2022079736W WO 2022188776 A1 WO2022188776 A1 WO 2022188776A1
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chr17
seq
combination
chr18
gastric cancer
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Chinese (zh)
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刘鑫
李慧
杨婷
陈志伟
范建兵
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广州市基准医疗有限责任公司
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    • 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
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/118—Prognosis of disease development
    • 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

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  • the invention belongs to the field of biotechnology, and in particular relates to a gene methylation marker or a combination and application thereof that can be used for the accompanying diagnosis of gastric cancer HER2 protein overexpression or gene amplification.
  • Gastric carcinoma GC is one of the most common malignant tumors. Although the incidence of gastric cancer has decreased worldwide in recent years, it is still the second most common tumor of cancer-related deaths. It is estimated that there are 930,000 new cases every year. The number of deaths exceeds 700,000, and there are large regional differences in the incidence rate, which is related to diet, lifestyle, economic status, and the prevalence of Helicobacter pylori infection. More than 2/3 of new cases and deaths occurred in developing countries, including East Asia. 42.4% (mainly China).
  • HER2 human epidermal growth factor receptor 2
  • HER2 protein can activate cell growth and proliferation-related signaling pathways.
  • Breast cancer, gastric cancer and other tumors are closely related.
  • companion diagnostics an in vitro diagnostic technology, has received increasing attention. This technology can be used to detect biomarkers and correlate with the use of specific drugs, and is an important part of the field of precise tumor diagnosis and treatment.
  • the HER2 companion diagnostic test for breast cancer has been widely used in the clinical diagnosis and treatment of breast cancer patients.
  • Liquid biopsy can obtain the ctDNA shed by the tumor and circulate in the blood by non-invasive blood drawing in vitro, so as to determine the gene mutation of the tumor. Therefore, the detection of HER2 gene copy number in gastric cancer patients by liquid biopsy instead of tissue biopsy will effectively alleviate the tissue sampling injury faced by gastric cancer patients, and can eliminate the problem of tissue heterogeneity through multiple blood sample collection, and improve the detection of HER2 positive gastric cancer. Accuracy and convenience.
  • cfDNA cell-free DNA
  • peripheral blood derived from the metabolism and apoptosis of normal cells or tumor cells, and contains genetic information such as somatic mutation and DNA methylation.
  • Liquid Biopsy the technology of mastering the occurrence and development of diseases. Compared with traditional tissue biopsy, it has the advantages of rapidity, convenience, less damage, and can be passed through multiple blood Sample collection eliminates the problem of tissue heterogeneity, improves the accuracy and convenience of HER2-positive gastric cancer detection, and many other advantages.
  • ctDNA is a sensitive and specific biomarker with broad applicability, which can be used for clinical and research of various types of cancers.
  • Professor Lu Yuming proved the technical and theoretical feasibility of liquid biopsy to replace tissue biopsy through cfDNA whole-genome methylation sequencing; in 2017, Professor Zhang Kun's team used ctDNA methylation to quantitatively describe tumor burden and tumor origin.
  • ctDNA profile In 2017, Professor Xu Ruihua's team found that by detecting the methylation level of specific sites of circulating tumor DNA (ctDNA) in a small amount (4-5ml) of peripheral blood, accurate early diagnosis of liver cancer and prediction of curative effect and prognosis can be made. Recently, more and more clinical studies have shown that plasma ctDNA methylation markers can be used as biomarkers for early diagnosis and screening of tumors, prediction, response to treatment, and monitoring tumor size and recurrence. At present, the international research direction is to integrate multi-omics/multi-molecular markers, multi-gene/multi-locus to improve the sensitivity and specificity of detection technology to meet the clinical demand for detection products.
  • One of the objectives of the present invention is to provide a plasma gene methylation marker and/or a combination thereof that can be used for the companion diagnosis of HER2 protein overexpression or gene amplification in gastric cancer.
  • a methylation marker or a combination thereof that can be used for the accompanying diagnosis of HER2 protein overexpression or gene amplification in gastric cancer, the methylation marker or a combination thereof comprising any one or a combination of two or more selected from the following: chr17:37879893, chr17:37881021, chr17:37886390, chr17:37893765, chr17:37860353, chr17:37843592.
  • Another object of the present invention is to provide the application of the above methylation markers or combinations thereof and/or detection-related reagents in the preparation of kits for detection, screening, prognosis monitoring, and companion diagnosis of gastric cancer HER2 protein overexpression or gene amplification .
  • Another object of the present invention is to provide a kit for detection, screening, prognosis monitoring, and companion diagnosis of gastric cancer HER2 protein overexpression or gene amplification.
  • the kit can be used for medication guidance for accompanying diagnosis of HER2 protein overexpression or gene amplification in gastric cancer.
  • Another object of the present invention is to provide a detection method for accompanying diagnosis of gastric cancer HER2, comprising the following steps: obtaining a sample to be detected;
  • the detection sample is subjected to the detection of the methylation marker or a combination thereof.
  • the present invention obtains genomic fragments (methylation markers chr17:37879893, chr17:37881021, chr17:37886390, chr17:37893765, chr17:37860353) that are modified by methylation in gastric cancer HER2+ and HER2- malignant patients, especially in plasma ⁇ ( ⁇ chr17:37862986, chr17:37862178,chr17:37884314,chr17:37879819,chr17:37895887,chr17:37893999,chr17:37855528,chr3:38080799,chr17:37843863,chr17:37821433,chr17:37843592 , chr17:37856695), the methylation markers provided by the present invention, for tissue and plasma, especially the combination of plasma genome fragments can be used as HER2R protein overexpression or gene amplification medication-guided companion diagnostic diagnostic markers, there are Companion
  • the present invention further provides a plurality of combined markers, such as chr17:37843592, chr17:37879893 and chr17:37860353, chr17:37843592, chr17:37879893 and chr17:37860353 and chr17:37843592, chr17:37862178 for use in Detection, screening, detection, prognosis monitoring and medication guidance of patients with IHC 2+GC.
  • Example 1 is a heat map of biomarkers screened by methylation level difference analysis between HER2+ gastric cancer tissue and HER2- gastric cancer tissue in Example 3.
  • Figure 2 is the performance of all methylation marker models in Example 3 on the test set and validation set in tissue samples.
  • Figure 3 shows the performance of the 10 methylation marker models in Example 4 on the test set and the validation set in tissue samples.
  • Figure 4 is the performance of all methylation marker models in Example 6 in the plasma independent validation set.
  • Figure 5 is the performance of the five methylation marker models in Example 6 in the plasma independent validation set.
  • Figure 6 is the performance of the three methylation marker models in Example 6 in the plasma independent validation set.
  • FIG. 7 is the performance of the two methylation marker models in Example 6 in the independent validation set of plasma.
  • Figure 8 is the performance of the three combined methylation marker models in Example 7 in the plasma independent validation set.
  • complementarity refers to a nucleotide (eg, 1 nucleotide) or a polynucleotide (eg, a sequence of nucleotides) related to the base pairing rules.
  • sequence 5'-A-G-T-3' is complementary to the sequence 3'-T-C-A-5'.
  • Complementarity can be “partial” in which only some nucleic acid bases are matched according to base pairing rules. Alternatively, there may be “complete” or “total” complementarity between nucleic acids. The degree of complementarity between nucleic acid strands affects the efficiency and strength of hybridization between nucleic acid strands. This is especially important in amplification reactions and detection methods that rely on binding between nucleic acids.
  • polymerase chain reaction is used to amplify target sequences, and the method consists of the steps of introducing a large excess of two oligonucleotide primers into a DNA mixture containing the desired target sequence, followed by the presence of DNA polymerase The precise thermal cycling sequence is performed below. Both primers are complementary to the corresponding strands of the double-stranded target sequence. For amplification, the mixture is denatured and the primers are then annealed to their complementary sequences within the target molecule. After annealing, the primers are amplified with a polymerase to form a new pair of complementary strands.
  • the steps of denaturation, primer annealing, and polymerase extension can be repeated multiple times (ie, denaturation, annealing, and extension constitute one "cycle”; there can be many "cycles") to obtain high concentrations of amplified fragments of the desired target sequence.
  • the length of the amplified fragment of the desired target sequence is determined by the relative position of the primers with respect to each other, so this length is a controllable parameter. Due to the repetitive aspect of the method, the method is referred to as the "polymerase chain reaction" ("PCR"). Since the desired amplified fragment of the target sequence becomes the predominant sequence (in terms of concentration) in the mixture, it is said to be “PCR amplified” and is the "PCR product" or "amplicon”.
  • nucleic acid detection refers to any method of determining the nucleotide composition of a target nucleic acid. Nucleic acid detection assays include, but are not limited to, DNA sequencing methods, probe hybridization methods.
  • amplifiable nucleic acid refers to a nucleic acid that can be amplified by any amplification method. It is expected that the "amplifiable nucleic acid” will generally comprise a "sample template”.
  • sample template refers to a nucleic acid derived from a sample for analysis of the presence of a "target” (defined below).
  • methylation refers to the methylation of cytosine at the C5 or N4 position of cytosine, the N6 position of adenine, or other types of nucleic acid methylation.
  • In vitro amplified DNA is usually unmethylated, as typically in vitro DNA amplification methods do not preserve the methylation pattern of the amplified template.
  • unmethylated DNA or “methylated DNA” can also refer to amplified DNA that is unmethylated or methylated, respectively, from the original template.
  • a "methylated nucleotide” or “methylated nucleotide base” refers to the presence of a methyl moiety on a nucleotide base, where the methyl moiety is not present in a typical putative nucleus in nucleotide bases.
  • cytosine does not contain a methyl moiety on its pyrimidine ring, but 5-methylcytosine contains a methyl moiety at the 5-position of its pyrimidine ring. Therefore, cytosine is not a methylated nucleotide and 5-methylcytosine is a methylated nucleotide.
  • thymine contains a methyl moiety at the 5-position of its pyrimidine ring; however, for purposes herein, thymine is not considered a methylated nucleotide when present in DNA because thymine is DNA typical nucleotide bases.
  • Methylation status can optionally be represented or indicated by a "methylation value” (eg, representing methylation frequency, fraction, ratio, percentage, etc.).
  • Methylation values can, for example, quantify the amount of intact nucleic acid present after restriction digestion with methylation-dependent restriction enzymes, or by comparing amplification profiles after bisulfite reaction, or by comparing bisulfite treatment and generated from untreated nucleic acid sequences.
  • values such as methylation values represent methylation status and thus can be used as quantitative indicators of methylation status in multiple copies of a locus.
  • the degree of co-methylation is represented or indicated by the methylation status of more than one methylation site. In a methylated region, when the methylation status of more than one methylation site is methylated is defined as co-methylation.
  • bisulfite reagent refers to a reagent comprising, in some embodiments, bisulfite, disulfite, hydrogen sulfite, or a combination thereof.
  • DNA treated with bisulfite reagent its unmethylated cytosine nucleotides will be converted to uracil, while methylated cytosine and other bases remain unchanged, so that for example CpG binuclear can be distinguished.
  • methylation assay or “methylation level detection” refers to any assay used to determine the methylation status of one or more CpG dinucleotide sequences within a nucleic acid sequence.
  • the CpG site represents a dinucleotide pair
  • the base guanine (G) follows cytosine (C)
  • CpG is an abbreviation for cytosine (C)-phosphate (p)-guanine (G) .
  • a HER2+ patient refers to a patient with an IHC result of 3+ and a pathology report as HER2-positive;
  • a HER2-patient refers to a patient with an IHC result of 0 or 1+ and a pathology report as HER2-negative.
  • a "marker” refers to a condition that can be used to indicate the expression of HER2 in a subject.
  • markers can be nucleic acid sequences, macromolecules, small molecules, etc., for example, nucleic acid sequences of a certain length, or nucleotides at one specific site or nucleotides at two specific sites.
  • the marker provided by the present invention refers to a CpG site of a patient that can be used to detect, predict or diagnose whether a subject is a gastric cancer patient with HER2.
  • gene amplification refers to a selective increase in the copy number of a specific gene (eg, HER2) while the copy number of other genes is not proportionally increased.
  • the present invention detects the expression variation of HER2 gene by measuring methylation markers through methylation detection technology, and discloses the combination and application of plasma gene methylation markers for HER2 accompanying diagnosis of gastric cancer. Specifically, the study obtained genomic fragments with significantly abnormal methylation modifications in the plasma of gastric cancer HER2+ and HER2- malignant patients, namely (the site is in the genome position, and Hg19 is the reference genome)
  • a methylation marker or a combination thereof that can be used for the companion diagnosis of HER2 protein overexpression or gene amplification in gastric cancer, the methylation marker or a combination thereof comprising a group selected from the following Any one or a combination of two or more: chr17:37879893, chr17:37881021, chr17:37886390, chr17:37893765, chr17:37860353, chr17:37843592.
  • the methylation marker comprises: chr17:37879893.
  • the methylation marker or combination thereof comprises chr17:37879893 and chr17:37860353.
  • the methylation marker or a combination thereof further comprises at least one selected from chr17:37862986, chr17:37862178, chr17:37884314, chr17:37879819, chr17:37895887, chr17:37893999 , chr17:37855528, chr3:38080799, chr17:37843863, chr17:37821433, chr17:37843592, chr17:37856695, chr17:37926322.
  • the combination of methylation markers includes chr17:37879893, chr17:37886390 and chr17:37893999.
  • the combination of methylation markers includes chr17:37879893, chr17:37886390, chr17:37893765, chr17:37862178, and chr17:37893999.
  • the methylation marker or a combination thereof further comprises at least one selected from the group consisting of: chr8:41309473, chr17:37843265, chr17:37843818, chr17:37843793, chr17:37886349, chr17: 37886390,chr17:37820452,chr17:37820496,chr17:37968912,chr17:37968904,chr17:37893804,chr17:37894352,chr17:37893005,chr17:37894334,chr17:37893813,chr17:37894439,chr17:37894258,chr17:37894288, chr17:37886321,chr17:37926309,chr17:37894165,chr17:37894469,chr17:37873266,chr17:37894233,chr17:
  • the methylation marker combination comprises chr17:37879893, chr17:37862178, chr17:37862986, chr17:37894352, chr17:37820452, chr17:37893765, chr17:37886349, chr17:37856695, chr17:37856695, 37843793, chr17:37886390.
  • the methylation marker combination comprises chr17:37843592, chr17:37879893, and chr17:37860353.
  • the methylation marker combination comprises chr17:37843592, chr17:37893999, and chr17:37860353.
  • the methylation marker combination comprises chr17:37843592, chr17:37862178.
  • the above-mentioned combination of methylation markers and/or related reagents for detection thereof can be used in the preparation of a kit for detecting IHC 2+ gastric cancer.
  • the methylation marker combination comprises chr17:37879893, chr17:37881021, chr17:37886390, chr17:37893765, chr17:37860353, chr17:37862986, chr17:37862178, chr17:37884314, chr17:37884314, 37879819,chr17:37895887,chr17:37893999,chr17:37855528,chr8:41309473,chr17:37843592,chr17:37843863,chr17:37843265,chr17:37843818,chr17:37843793,chr17:37886349,chr17:37886390,chr17:37820452, chr17:37820496,chr17:37968912,chr17:37968904,chr17:37893804,chr1717:37879893
  • methylation markers or combinations thereof and/or detection-related reagents can be used in the preparation of kits for detection, screening, prognosis monitoring, and companion diagnosis of gastric cancer overexpression of HER2 protein or gene amplification.
  • kits for detection, screening, prognosis monitoring, and companion diagnosis of gastric cancer HER2 protein overexpression or gene amplification In some embodiments of the present invention, it relates to a kit for detection, screening, prognosis monitoring, and companion diagnosis of gastric cancer HER2 protein overexpression or gene amplification.
  • the kit can be used for medication guidance for accompanying diagnosis of HER2 protein overexpression or gene amplification in gastric cancer.
  • kits comprise reagents for detecting, screening, prognostic monitoring, companion diagnostics, methylation levels of the aforementioned methylation markers, or a combination thereof.
  • HM450K Illumina's Infinium HumanMethylation450BeadChip
  • HM450K Infinium CytoSNP- 850K BeadChip
  • Affymetrix any custom designed array
  • Sequencing methods may include, but are not limited to: Sanger sequencing, high-throughput sequencing, pyrosequencing, sequencing-by-synthesis, single-molecule sequencing, nanopore sequencing, semiconductor sequencing, sequencing by ligation, sequencing by hybridization, digital gene expression (Helicos), next-generation sequencing , Single Molecule Sequencing by Synthesis (SMSS) (Helicos), Massively Parallel Sequencing, Clonal Single Molecule Arrays (Solexa/Illumina), Shotgun Sequencing, Maxim Gilbert Sequencing, Primer Walking, Using PacBio, SOLiD, Ion Torrent or Nanopore Platforms Sequencing and any other sequencing method known in the art.
  • the kit adopts real-time fluorescent quantitative PCRR or other types of PCR detection methods, including the following primers and probes:
  • the sample detected by the kit is tissue or plasma or gastric juice, more preferably plasma.
  • ctDNA Screening markers for the diagnosis of gastric cancer based on the methylation of circulating tumor DNA (ctDNA), including the following steps:
  • Step 1 Perform methylation library building, targeted hybridization capture and next-generation sequencing in advanced gastric cancer tissue samples to initially screen potential methylation marker targets related to HER2 gene expression;
  • Step 2 Design and establish a corresponding methylation-specific qPCR (real-time quantitative PCR) technology platform for these targets;
  • Step 3 In gastric cancer plasma samples, screen and filter these targets, and find a marker set for distinguishing HER2 negative and positive;
  • Step 4 Use the marker set selected in step 6 of the above procedure to further screen markers in the plasma training set and validation set, build an algorithm model, and conduct independent data set validation;
  • Step 5 Confirm the gene methylation markers and algorithm models that are finally used for the diagnosis of advanced HER2 in gastric cancer.
  • This embodiment discloses the above-mentioned NGS-based detection method for differential methylation markers (markers) for the accompanying diagnosis of HER2R protein overexpression or gene amplification in gastric cancer patients, which specifically includes the following steps: 1. Tissue sample processing
  • tissue samples from patients with gastric cancer After clinical surgical resection of the tissue samples from patients with gastric cancer, they were fixed with 4% neutral formalin, dehydrated with gradient alcohol, cleared with xylene, immersed in wax, embedded in sections, and finally formed FFPE samples from patients with gastric cancer.
  • tissue DNA extraction was carried out according to the instructions of Qiagen's AllPre DNA/RNA FFPE Kit.
  • Target BP Peak
  • target BP target BP
  • the extracted tissue DNA is subjected to bisulfite conversion, so that the unmethylated cytosine in the DNA is deaminated into uracil, while the methylated cytosine remains unchanged to obtain sulfite
  • the specific operation of the conversion is carried out according to the instructions of Zymo Research's EZ DNA Methylation-Lightning Kit.
  • the present embodiment discloses the above-mentioned qPCR-based detection method for the differential methylation marker (marker) used for the HER2 companion diagnosis of gastric cancer patients, which specifically includes the following steps:
  • EDTAK2 anticoagulant vacuum blood collection tube (BD, Cat#367525) to collect 10 mL of whole blood, mix thoroughly to avoid hemolysis, and perform plasma separation processing on whole blood within 4-6 hours. 4°C, 1600g, 15min centrifugation, carefully suck the upper plasma, avoid sucking the middle buffy coat, the obtained plasma is centrifuged again at 4°C, 16000g, 10min in a high-speed centrifuge to obtain the desired sample plasma.
  • reaction system single dose ⁇ L 2*ChamQ Geno-SNP probe Master Mix 10 Primer F/R+probe (single marker) 2 DEPC H 2 O 2
  • This example discloses a specific biomarker for HER2 companion diagnosis of gastric cancer, using tissue samples from 44 HER2+ gastric cancer patients and 30 HER2- gastric cancer patients, using the method described in Example 1 (NGS- BASED) for methylation analysis of tissue samples.
  • Biomarkers related to HER2 expression were screened out using the methylation level differences in different groups, and the locus data were clustered in normal and gastric cancer samples. The results are shown in Figure 1.
  • a total of 102 sites with the most significant abnormal methylation in HER2+ and HER2- gastric cancer samples were screened (FDR ⁇ 0.01).
  • the red is the hypermethylated expression of the site
  • the blue is the hypomethylated expression of the site.
  • the 102 methylation biomarkers (hereinafter referred to as sites or markers) are:
  • the random forest model is used for modeling analysis, and the split is 6:4, and 20 repetitions are performed to obtain a model validation set AUC of 0.95, as shown in Figure 2.
  • an independent validation set namely 31 HER2+ gastric cancer patient tissue samples and 42 HER2- gastric cancer patient tissue samples were used to detect and judge , using the random forest model for modeling analysis, according to the 6:4 split, and repeating 20 times, the AUC of the model validation set is as high as 0.94, and the results are shown in Figure 3. It shows that the combination of 10 methylation markers can distinguish HER2+ and HER2- tissue samples well.
  • the methylation level analysis of tissue samples was carried out by using the method described in Example 2.
  • the independent plasma training set that is, the plasma samples of 7 HER2+ gastric cancer patients and the plasma samples of 20 HER2- gastric cancer patients were detected and judged.
  • the AUC of locus discrimination can be as high as 0.981, and the AUC of 5 marker loci (chr17:37879893, chr17:37881021, chr17:37886390, chr17:37893765, chr17:37860353) can be greater than 0.8, see Table 2 for details.
  • Example 5 Using all the markers in Example 5, the random forest model was used to model the plasma samples of 7 patients with HER2+ gastric cancer and the plasma samples of 20 patients with HER22- gastric cancer. According to the detection method described in Example 2, 3 patients with HER2+ were used for modeling. It was verified with 14 HER-independent plasma samples, and the judgment was completely correct, as shown in Figure 4.
  • a model was constructed using a combination of 5 markers (chr17:37879893, chr17:37886390, chr17:37893765, chr17:37862178, and chr17:37893999), and the detection method described in Example 2 was performed in 3 cases of HER2+ and 14 cases of HER2-independent
  • the plasma sample set was validated, and only one case of HER2- was misjudged, see Figure 5.
  • the results please See Table 7.2 and Figure 8. It shows that the combination of methylation markers can well distinguish FISH+ and FISH- tissue specimens in IHC2+ subgroup.

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Abstract

La présente invention concerne un marqueur de méthylation génique ou une combinaison associée pouvant être utilisés pour le diagnostic compagnon du carcinome gastrique HER2. Le marqueur de méthylation ou la combinaison associée comprend l'un quelconque ou une combinaison de deux ou plus de deux marqueurs choisis parmi les marqueurs suivants : chr17:37879893, chr17:37881021, chr17:37886390, chr17:37893765, chr17:37860353 et chr17:37843592. La présente invention concerne également un kit de détection et un procédé de détection pour un marqueur moléculaire de méthylation ou une combinaison associée.
PCT/CN2022/079736 2021-03-09 2022-03-08 Marqueur de méthylation génique ou combinaison associée pouvant être utilisés pour le diagnostic compagnon du carcinome gastrique her2, et son utilisation WO2022188776A1 (fr)

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Citations (4)

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