CN116219020B - 一种甲基化内参基因及其应用 - Google Patents
一种甲基化内参基因及其应用 Download PDFInfo
- Publication number
- CN116219020B CN116219020B CN202310227845.3A CN202310227845A CN116219020B CN 116219020 B CN116219020 B CN 116219020B CN 202310227845 A CN202310227845 A CN 202310227845A CN 116219020 B CN116219020 B CN 116219020B
- Authority
- CN
- China
- Prior art keywords
- gene
- seq
- methylation
- dna
- sdf4
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 238
- 230000011987 methylation Effects 0.000 title claims abstract description 96
- 238000007069 methylation reaction Methods 0.000 title claims abstract description 96
- 238000001514 detection method Methods 0.000 claims abstract description 31
- 201000010099 disease Diseases 0.000 claims abstract description 21
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 21
- 239000000523 sample Substances 0.000 claims description 152
- 210000001519 tissue Anatomy 0.000 claims description 72
- 210000002381 plasma Anatomy 0.000 claims description 71
- 201000007270 liver cancer Diseases 0.000 claims description 62
- 208000014018 liver neoplasm Diseases 0.000 claims description 62
- 238000006243 chemical reaction Methods 0.000 claims description 61
- 239000012634 fragment Substances 0.000 claims description 42
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 34
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 claims description 29
- 108091029430 CpG site Proteins 0.000 claims description 24
- 239000003153 chemical reaction reagent Substances 0.000 claims description 24
- 208000019425 cirrhosis of liver Diseases 0.000 claims description 20
- 241000282414 Homo sapiens Species 0.000 claims description 16
- 210000004027 cell Anatomy 0.000 claims description 15
- 239000002773 nucleotide Substances 0.000 claims description 15
- 125000003729 nucleotide group Chemical group 0.000 claims description 15
- 238000011144 upstream manufacturing Methods 0.000 claims description 13
- 108020004414 DNA Proteins 0.000 claims description 9
- 108700039691 Genetic Promoter Regions Proteins 0.000 claims description 8
- 210000004369 blood Anatomy 0.000 claims description 5
- 239000008280 blood Substances 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 230000001105 regulatory effect Effects 0.000 claims description 5
- 206010003445 Ascites Diseases 0.000 claims description 4
- 206010048612 Hydrothorax Diseases 0.000 claims description 4
- 206010036790 Productive cough Diseases 0.000 claims description 4
- 210000001175 cerebrospinal fluid Anatomy 0.000 claims description 4
- 210000000349 chromosome Anatomy 0.000 claims description 4
- 238000003745 diagnosis Methods 0.000 claims description 4
- 210000003296 saliva Anatomy 0.000 claims description 4
- 210000002966 serum Anatomy 0.000 claims description 4
- 210000003802 sputum Anatomy 0.000 claims description 4
- 208000024794 sputum Diseases 0.000 claims description 4
- 210000002700 urine Anatomy 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 3
- 230000007613 environmental effect Effects 0.000 claims description 2
- 239000003550 marker Substances 0.000 claims description 2
- 101000899406 Homo sapiens 45 kDa calcium-binding protein Proteins 0.000 abstract description 80
- 230000003321 amplification Effects 0.000 abstract description 52
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 52
- 206010028980 Neoplasm Diseases 0.000 abstract description 37
- 238000000034 method Methods 0.000 abstract description 29
- 238000013399 early diagnosis Methods 0.000 abstract description 8
- 238000012216 screening Methods 0.000 abstract description 8
- 238000007855 methylation-specific PCR Methods 0.000 abstract description 7
- 230000008569 process Effects 0.000 abstract description 4
- 238000012544 monitoring process Methods 0.000 abstract description 3
- 230000002980 postoperative effect Effects 0.000 abstract description 3
- 238000012502 risk assessment Methods 0.000 abstract 1
- 101000756632 Homo sapiens Actin, cytoplasmic 1 Proteins 0.000 description 66
- 102100022530 45 kDa calcium-binding protein Human genes 0.000 description 43
- 201000011510 cancer Diseases 0.000 description 29
- 101000976642 Homo sapiens Zinc finger protein ZIC 4 Proteins 0.000 description 21
- 206010006187 Breast cancer Diseases 0.000 description 17
- 208000026310 Breast neoplasm Diseases 0.000 description 17
- 230000004544 DNA amplification Effects 0.000 description 17
- CTMZLDSMFCVUNX-VMIOUTBZSA-N cytidylyl-(3'->5')-guanosine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@H](OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=C(C(N=C(N)N3)=O)N=C2)O)[C@@H](CO)O1 CTMZLDSMFCVUNX-VMIOUTBZSA-N 0.000 description 17
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 16
- 201000005202 lung cancer Diseases 0.000 description 16
- 208000020816 lung neoplasm Diseases 0.000 description 16
- 210000000265 leukocyte Anatomy 0.000 description 14
- 238000012795 verification Methods 0.000 description 13
- 238000000605 extraction Methods 0.000 description 11
- 101150051644 SDF4 gene Proteins 0.000 description 10
- 238000012163 sequencing technique Methods 0.000 description 10
- 108091029523 CpG island Proteins 0.000 description 7
- 230000007067 DNA methylation Effects 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 102100023493 Zinc finger protein ZIC 4 Human genes 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 230000014509 gene expression Effects 0.000 description 7
- 239000002096 quantum dot Substances 0.000 description 7
- UDGUGZTYGWUUSG-UHFFFAOYSA-N 4-[4-[[2,5-dimethoxy-4-[(4-nitrophenyl)diazenyl]phenyl]diazenyl]-n-methylanilino]butanoic acid Chemical compound COC=1C=C(N=NC=2C=CC(=CC=2)N(C)CCCC(O)=O)C(OC)=CC=1N=NC1=CC=C([N+]([O-])=O)C=C1 UDGUGZTYGWUUSG-UHFFFAOYSA-N 0.000 description 6
- 210000000481 breast Anatomy 0.000 description 6
- 241000894007 species Species 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 201000009030 Carcinoma Diseases 0.000 description 5
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 5
- 101000937544 Homo sapiens Beta-2-microglobulin Proteins 0.000 description 5
- 206010039491 Sarcoma Diseases 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 5
- 238000007689 inspection Methods 0.000 description 5
- 210000000056 organ Anatomy 0.000 description 5
- 102000053602 DNA Human genes 0.000 description 4
- 101100545265 Homo sapiens ZIC4 gene Proteins 0.000 description 4
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 4
- 238000003149 assay kit Methods 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 239000003623 enhancer Substances 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 238000001114 immunoprecipitation Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000003020 moisturizing effect Effects 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 3
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 3
- 208000030808 Clear cell renal carcinoma Diseases 0.000 description 3
- 206010014733 Endometrial cancer Diseases 0.000 description 3
- 206010014759 Endometrial neoplasm Diseases 0.000 description 3
- 208000032320 Germ cell tumor of testis Diseases 0.000 description 3
- 201000010915 Glioblastoma multiforme Diseases 0.000 description 3
- 206010025323 Lymphomas Diseases 0.000 description 3
- 206010027406 Mesothelioma Diseases 0.000 description 3
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 3
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 3
- 206010061332 Paraganglion neoplasm Diseases 0.000 description 3
- 206010060862 Prostate cancer Diseases 0.000 description 3
- 208000000102 Squamous Cell Carcinoma of Head and Neck Diseases 0.000 description 3
- 208000034254 Squamous cell carcinoma of the cervix uteri Diseases 0.000 description 3
- 208000024770 Thyroid neoplasm Diseases 0.000 description 3
- 201000005969 Uveal melanoma Diseases 0.000 description 3
- 238000001369 bisulfite sequencing Methods 0.000 description 3
- 206010005084 bladder transitional cell carcinoma Diseases 0.000 description 3
- 201000006612 cervical squamous cell carcinoma Diseases 0.000 description 3
- 208000006990 cholangiocarcinoma Diseases 0.000 description 3
- 206010073251 clear cell renal cell carcinoma Diseases 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 201000010897 colon adenocarcinoma Diseases 0.000 description 3
- 208000029742 colonic neoplasm Diseases 0.000 description 3
- 208000030381 cutaneous melanoma Diseases 0.000 description 3
- -1 cy5 Chemical compound 0.000 description 3
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 201000006585 gastric adenocarcinoma Diseases 0.000 description 3
- 208000005017 glioblastoma Diseases 0.000 description 3
- 201000000459 head and neck squamous cell carcinoma Diseases 0.000 description 3
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 3
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 3
- 230000000977 initiatory effect Effects 0.000 description 3
- 208000030173 low grade glioma Diseases 0.000 description 3
- 201000005249 lung adenocarcinoma Diseases 0.000 description 3
- 201000005243 lung squamous cell carcinoma Diseases 0.000 description 3
- 201000001441 melanoma Diseases 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 201000010302 ovarian serous cystadenocarcinoma Diseases 0.000 description 3
- 208000008443 pancreatic carcinoma Diseases 0.000 description 3
- 208000007312 paraganglioma Diseases 0.000 description 3
- 208000028591 pheochromocytoma Diseases 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- 201000003708 skin melanoma Diseases 0.000 description 3
- 208000002918 testicular germ cell tumor Diseases 0.000 description 3
- 201000002510 thyroid cancer Diseases 0.000 description 3
- 206010046766 uterine cancer Diseases 0.000 description 3
- 208000012991 uterine carcinoma Diseases 0.000 description 3
- SGTNSNPWRIOYBX-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-{[2-(3,4-dimethoxyphenyl)ethyl](methyl)amino}-2-(propan-2-yl)pentanenitrile Chemical compound C1=C(OC)C(OC)=CC=C1CCN(C)CCCC(C#N)(C(C)C)C1=CC=C(OC)C(OC)=C1 SGTNSNPWRIOYBX-UHFFFAOYSA-N 0.000 description 2
- 101710168918 45 kDa calcium-binding protein Proteins 0.000 description 2
- 206010001197 Adenocarcinoma of the cervix Diseases 0.000 description 2
- 208000034246 Adenocarcinoma of the cervix uteri Diseases 0.000 description 2
- 108091093088 Amplicon Proteins 0.000 description 2
- 108010045403 Calcium-Binding Proteins Proteins 0.000 description 2
- 102000005701 Calcium-Binding Proteins Human genes 0.000 description 2
- 108010009540 DNA (Cytosine-5-)-Methyltransferase 1 Proteins 0.000 description 2
- 102100036279 DNA (cytosine-5)-methyltransferase 1 Human genes 0.000 description 2
- 108010042407 Endonucleases Proteins 0.000 description 2
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 2
- 108700039887 Essential Genes Proteins 0.000 description 2
- 206010064571 Gene mutation Diseases 0.000 description 2
- 108060004795 Methyltransferase Proteins 0.000 description 2
- 102000016397 Methyltransferase Human genes 0.000 description 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- 108091008109 Pseudogenes Proteins 0.000 description 2
- 102000057361 Pseudogenes Human genes 0.000 description 2
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 2
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 2
- 239000000090 biomarker Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 201000006662 cervical adenocarcinoma Diseases 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 230000000875 corresponding effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 201000003914 endometrial carcinoma Diseases 0.000 description 2
- 238000001976 enzyme digestion Methods 0.000 description 2
- 201000004101 esophageal cancer Diseases 0.000 description 2
- 230000030279 gene silencing Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000006607 hypermethylation Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 238000001543 one-way ANOVA Methods 0.000 description 2
- 201000002528 pancreatic cancer Diseases 0.000 description 2
- 238000010791 quenching Methods 0.000 description 2
- 230000000171 quenching effect Effects 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 210000002536 stromal cell Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229940035893 uracil Drugs 0.000 description 2
- 108020004463 18S ribosomal RNA Proteins 0.000 description 1
- WCKQPPQRFNHPRJ-UHFFFAOYSA-N 4-[[4-(dimethylamino)phenyl]diazenyl]benzoic acid Chemical compound C1=CC(N(C)C)=CC=C1N=NC1=CC=C(C(O)=O)C=C1 WCKQPPQRFNHPRJ-UHFFFAOYSA-N 0.000 description 1
- 101150087690 ACTB gene Proteins 0.000 description 1
- WNDDWSAHNYBXKY-UHFFFAOYSA-N ATTO 425-2 Chemical compound CC1CC(C)(C)N(CCCC(O)=O)C2=C1C=C1C=C(C(=O)OCC)C(=O)OC1=C2 WNDDWSAHNYBXKY-UHFFFAOYSA-N 0.000 description 1
- PWZJEXGKUHVUFP-UHFFFAOYSA-N ATTO 590 meta-isomer Chemical compound [O-]Cl(=O)(=O)=O.C1=2C=C3C(C)=CC(C)(C)N(CC)C3=CC=2OC2=CC3=[N+](CC)C(C)(C)C=C(C)C3=CC2=C1C1=CC=C(C(O)=O)C=C1C(O)=O PWZJEXGKUHVUFP-UHFFFAOYSA-N 0.000 description 1
- 208000006468 Adrenal Cortex Neoplasms Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 208000017897 Carcinoma of esophagus Diseases 0.000 description 1
- 102100031780 Endonuclease Human genes 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 208000031448 Genomic Instability Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 208000033781 Thyroid carcinoma Diseases 0.000 description 1
- GYDJEQRTZSCIOI-UHFFFAOYSA-N Tranexamic acid Chemical compound NCC1CCC(C(O)=O)CC1 GYDJEQRTZSCIOI-UHFFFAOYSA-N 0.000 description 1
- 208000034953 Twin anemia-polycythemia sequence Diseases 0.000 description 1
- 208000020990 adrenal cortex carcinoma Diseases 0.000 description 1
- 230000001919 adrenal effect Effects 0.000 description 1
- 208000007128 adrenocortical carcinoma Diseases 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 201000001528 bladder urothelial carcinoma Diseases 0.000 description 1
- NNTOJPXOCKCMKR-UHFFFAOYSA-N boron;pyridine Chemical compound [B].C1=CC=NC=C1 NNTOJPXOCKCMKR-UHFFFAOYSA-N 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 210000003679 cervix uteri Anatomy 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 230000001054 cortical effect Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000019975 dosage compensation by inactivation of X chromosome Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 201000003683 endocervical adenocarcinoma Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000004049 epigenetic modification Effects 0.000 description 1
- 201000005619 esophageal carcinoma Diseases 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 208000021045 exocrine pancreatic carcinoma Diseases 0.000 description 1
- 210000001508 eye Anatomy 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 239000002122 magnetic nanoparticle Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 210000001331 nose Anatomy 0.000 description 1
- BRJCLSQFZSHLRL-UHFFFAOYSA-N oregon green 488 Chemical compound OC(=O)C1=CC(C(=O)O)=CC=C1C1=C2C=C(F)C(=O)C=C2OC2=CC(O)=C(F)C=C21 BRJCLSQFZSHLRL-UHFFFAOYSA-N 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- VYNDHICBIRRPFP-UHFFFAOYSA-N pacific blue Chemical compound FC1=C(O)C(F)=C2OC(=O)C(C(=O)O)=CC2=C1 VYNDHICBIRRPFP-UHFFFAOYSA-N 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 201000001514 prostate carcinoma Diseases 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- ABZLKHKQJHEPAX-UHFFFAOYSA-N tetramethylrhodamine Chemical compound C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C([O-])=O ABZLKHKQJHEPAX-UHFFFAOYSA-N 0.000 description 1
- MPLHNVLQVRSVEE-UHFFFAOYSA-N texas red Chemical compound [O-]S(=O)(=O)C1=CC(S(Cl)(=O)=O)=CC=C1C(C1=CC=2CCCN3CCCC(C=23)=C1O1)=C2C1=C(CCC1)C3=[N+]1CCCC3=C2 MPLHNVLQVRSVEE-UHFFFAOYSA-N 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 208000013077 thyroid gland carcinoma Diseases 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
Classifications
-
- 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
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4728—Calcium binding proteins, e.g. calmodulin
-
- 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
- C12Q2600/118—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
-
- 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
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Biophysics (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- Physics & Mathematics (AREA)
- Toxicology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
试剂组分 | 终浓度 |
10×PCR Buffer | 1× |
4×Enhancer | 1× |
MgCl2(250mM) | 6mM |
dNTP Mix(25mM each) | 350μM |
Taq酶(5U/μL) | 3U |
引物对1 | 0.2μM |
探针2 | 200nM |
DNA模板3 | 0-200ng |
无核酸酶水 | 补水到25μL |
总体积 | 25μL |
模板起始量 | SDF4(引物探针组合5) | SDF4(引物探针组合6) | ACTB |
0ng | Undetermined | Undetermined | Undetermined |
5ng | 14.41 | 14.53 | 16.90 |
10ng | 13.21 | 13.50 | 15.70 |
20ng | 12.08 | 12.36 | 14.28 |
30ng | 11.40 | 11.74 | 13.70 |
50ng | 10.60 | 10.97 | 13.07 |
100ng | 10.08 | 10.41 | 12.62 |
200ng | 9.23 | 9.55 | 11.92 |
试剂组分 | 终浓度 |
10×PCR Buffer | 1× |
4×Enhancer | 1× |
MgCl2(250mM) | 6mM |
dNTP Mix(25mM each) | 350μM |
Taq酶(5U/μL) | 3U |
ZIC4引物对(5μM) | 0.2μM |
ZIC4探针(10μM) | 250nM |
内参基因引物对(5μM) | 0.2μM |
内参基因探针(10μM) | 250nM |
DNA模板 | 10ng |
无核酸酶水 | 补水至25μL |
总体积 | 25μL |
试剂组分 | 终浓度 |
10×PCR Buffer | 1× |
4×Enhancer | 1× |
MgCl2(250mM) | 6mM |
dNTP Mix(25mM each) | 350μM |
Taq酶(5U/μL) | 3U |
ZIC4引物对(5μM) | 0.2μM |
ZIC4探针(10μM) | 250nM |
内参基因引物对(5μM) | 0.2因引 |
内参基因探针(10μM) | 250nM |
DNA模板 | 10ng |
无核酸酶水 | 补水至25μL |
总体积 | 25μL |
序号 | 样本类别 | cfDNA浓度(ng/μL) |
1 | 肝癌血浆 | 10.200 |
2 | 肝癌血浆 | 7.640 |
3 | 肝癌血浆 | 4.500 |
4 | 肝癌血浆 | 3.320 |
5 | 肝癌血浆 | 3.120 |
6 | 肝癌血浆 | 2.520 |
7 | 肝癌血浆 | 2.420 |
8 | 肝癌血浆 | 1.790 |
9 | 肝癌血浆 | 1.740 |
10 | 肝癌血浆 | 1.740 |
11 | 肝癌血浆 | 1.710 |
12 | 肝癌血浆 | 1.490 |
13 | 肝癌血浆 | 1.450 |
14 | 肝硬化血浆 | 1.700 |
15 | 肝硬化血浆 | 1.460 |
16 | 肝硬化血浆 | 1.400 |
17 | 产前血浆 | 0.390 |
18 | 产前血浆 | 0.386 |
19 | 产前血浆 | 0.356 |
20 | 产前血浆 | 0.338 |
21 | 产前血浆 | 0.338 |
22 | 产前血浆 | 0.352 |
23 | 产前血浆 | 0.348 |
24 | 产前血浆 | 0.378 |
25 | 产前血浆 | 0.376 |
26 | 产前血浆 | 0.382 |
试剂组分 | 终浓度 |
10×PCR Buffer | 1× |
4×Enhancer | 1× |
MgCl2(250mM) | 6mM |
dNTP Mix(25mM each) | 350μM |
Taq酶(5U/μL) | 3U |
靶基因引物对(5μM) | 0.2μM |
靶基因探针(10μM) | 200nM |
ACTB引物对(5μM) | 0.3μM |
ACTB探针(10μM) | 200nM |
SDF4引物对(5μM) | 0.2μM |
SDF4探针(10μM) | 100nM |
转化DNA模板 | / |
无核酸酶水 | 补水至25μL |
总体积 | 25μL |
Claims (14)
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310227845.3A CN116219020B (zh) | 2023-03-02 | 2023-03-02 | 一种甲基化内参基因及其应用 |
PCT/CN2023/105352 WO2024178886A1 (zh) | 2023-03-02 | 2023-06-30 | 一种甲基化内参基因及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310227845.3A CN116219020B (zh) | 2023-03-02 | 2023-03-02 | 一种甲基化内参基因及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN116219020A CN116219020A (zh) | 2023-06-06 |
CN116219020B true CN116219020B (zh) | 2023-11-07 |
Family
ID=86588958
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310227845.3A Active CN116219020B (zh) | 2023-03-02 | 2023-03-02 | 一种甲基化内参基因及其应用 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN116219020B (zh) |
WO (1) | WO2024178886A1 (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116219020B (zh) * | 2023-03-02 | 2023-11-07 | 北京和瑞精湛医学检验实验室有限公司 | 一种甲基化内参基因及其应用 |
CN116555432B (zh) * | 2023-07-05 | 2023-09-05 | 广州凯普医药科技有限公司 | 一种膀胱癌快速检测试剂盒 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102027118A (zh) * | 2008-04-30 | 2011-04-20 | 桑比欧公司 | 含有dna甲基化状态改变的神经再生细胞 |
CN106636390A (zh) * | 2016-12-16 | 2017-05-10 | 江苏为真生物医药技术股份有限公司 | 基因甲基化检测方法和检测试剂盒及其应用 |
CN110387421A (zh) * | 2019-08-28 | 2019-10-29 | 深圳市新合生物医疗科技有限公司 | 用于肺癌检测的DNA甲基化qPCR试剂盒及使用方法 |
CN112760374A (zh) * | 2021-03-23 | 2021-05-07 | 浙江大学 | Sdf4作为生物标志物在脓毒症预后预测试剂中的应用 |
JP2022026407A (ja) * | 2020-07-31 | 2022-02-10 | 花王株式会社 | プラセボ効果を評価する方法 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101007926B1 (ko) * | 2006-12-27 | 2011-01-12 | (주)레퍼런스바이오랩 | 표준 발현 유전자를 발굴하기 위한 유전자 발현 데이터처리, 분석 방법 |
CN109385465B (zh) * | 2018-07-27 | 2019-12-24 | 中山大学附属第六医院 | 一种dna甲基化定量系统 |
US20210335451A1 (en) * | 2020-04-24 | 2021-10-28 | Augusta University Research Institute, Inc. | Transcriptomic signature for the prognosis and treatment selection for cervical cancer |
CN116219020B (zh) * | 2023-03-02 | 2023-11-07 | 北京和瑞精湛医学检验实验室有限公司 | 一种甲基化内参基因及其应用 |
-
2023
- 2023-03-02 CN CN202310227845.3A patent/CN116219020B/zh active Active
- 2023-06-30 WO PCT/CN2023/105352 patent/WO2024178886A1/zh unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102027118A (zh) * | 2008-04-30 | 2011-04-20 | 桑比欧公司 | 含有dna甲基化状态改变的神经再生细胞 |
CN106636390A (zh) * | 2016-12-16 | 2017-05-10 | 江苏为真生物医药技术股份有限公司 | 基因甲基化检测方法和检测试剂盒及其应用 |
CN110387421A (zh) * | 2019-08-28 | 2019-10-29 | 深圳市新合生物医疗科技有限公司 | 用于肺癌检测的DNA甲基化qPCR试剂盒及使用方法 |
JP2022026407A (ja) * | 2020-07-31 | 2022-02-10 | 花王株式会社 | プラセボ効果を評価する方法 |
CN112760374A (zh) * | 2021-03-23 | 2021-05-07 | 浙江大学 | Sdf4作为生物标志物在脓毒症预后预测试剂中的应用 |
Non-Patent Citations (3)
Title |
---|
"SDF-1及其受体CXCR4在哮喘大鼠肺组织中的表达及AMD3100的干预作用";王利霞;《中国学位论文全文数据库》;摘要 * |
"不同细胞系和组织基因的甲基化差异分析及其序列与CpG甲基化程度的关联";艾雪;《CNKI硕士电子期刊》;摘要 * |
Gregory,S.G.等."Homo sapiens chromosome 1,GRCh38.p14 Primary Assembly;NCBI Reference Sequence: NC_000001.11".《GenBank》.2022,第1-12页. * |
Also Published As
Publication number | Publication date |
---|---|
CN116219020A (zh) | 2023-06-06 |
WO2024178886A1 (zh) | 2024-09-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10450609B2 (en) | Quantitative multiplex methylation specific PCR method—cMethDNA, reagents, and its use | |
EP2885427B1 (en) | Colorectal cancer methylation marker | |
CN116219020B (zh) | 一种甲基化内参基因及其应用 | |
EP3524688B1 (en) | Multiple detection method of methylated dna | |
JP2022525890A (ja) | Dna試料のメチル化変化を検出するための方法およびシステム | |
US9540697B2 (en) | Prostate cancer markers | |
CA2775671A1 (en) | Method for analysis of dna methylation profiles of cell-free circulating dna in bodily fluids | |
CN114891886B (zh) | 用于诊断膀胱癌的核酸产品、试剂盒及应用 | |
WO2021185061A1 (en) | Methods and kits for screening colorectal neoplasm | |
US20190032143A1 (en) | Kits and methods for diagnosis, screening, treatment and disease monitoring | |
CN116200499B (zh) | 一种用于肝癌检测的基因组合与相关试剂和应用 | |
JP2023524067A (ja) | ヌクレアーゼ、ライゲーション、脱アミノ化、dna修復、およびポリメラーゼ反応と、キャリーオーバー防止との組み合わせを用いた、核酸配列、変異、コピー数、またはメチル化変化の特定および相対的定量化のための方法およびマーカー | |
CN113493835A (zh) | 通过检测bcan基因区域的甲基化状态筛查大肠瘤的方法和试剂盒 | |
WO2022170984A1 (zh) | 结直肠进展期腺瘤的筛查、风险评估及预后方法和试剂盒 | |
CN117821585A (zh) | 结直肠癌早期诊断标志物及应用 | |
CN112430657B (zh) | 结直肠癌相关的甲基化标志物以及用于检测结直肠癌的试剂盒 | |
WO2020221314A1 (zh) | 基于甲基化修饰的肿瘤标记物stamp-ep7及其应用 | |
CN115094139B (zh) | 检测甲基化水平的试剂在制备膀胱癌诊断产品中的应用以及膀胱癌诊断试剂盒 | |
WO2023104136A1 (zh) | 甲状腺癌良恶性结节诊断的甲基化标志物及其应用 | |
CN117512115A (zh) | 一种通过基因甲基化信号预放大提高检测肿瘤灵敏率的方法 | |
TW202328459A (zh) | 一種腫瘤檢測方法及應用 | |
WO2023135600A1 (en) | Personalized cancer management and monitoring based on dna methylation changes in cell-free dna | |
CN116516005A (zh) | 一种用于检测头颈鳞癌的核酸产品、试剂盒及应用 | |
KR20230105973A (ko) | 특정 유전자의 CpG 메틸화 변화를 이용한 전립선암 진단용 조성물 및 이의 용도 | |
CN114634982A (zh) | 一种检测多核苷酸变异的方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CB03 | Change of inventor or designer information | ||
CB03 | Change of inventor or designer information |
Inventor after: Li Hui Inventor after: Yang Hao Inventor after: Zhang Aiyuan Inventor after: Xi Liying Inventor after: Zheng Lu Inventor after: Wang Yin Inventor after: Bai Jian Inventor after: Wu Lin Inventor before: Li Hui Inventor before: Yang Hao Inventor before: Zhang Aiyuan Inventor before: Xi Liying Inventor before: Zheng Lu Inventor before: Wang Yin Inventor before: Bai Jian Inventor before: Wu Lin |