JP5535921B2 - 核酸増幅 - Google Patents
核酸増幅 Download PDFInfo
- Publication number
- JP5535921B2 JP5535921B2 JP2010527295A JP2010527295A JP5535921B2 JP 5535921 B2 JP5535921 B2 JP 5535921B2 JP 2010527295 A JP2010527295 A JP 2010527295A JP 2010527295 A JP2010527295 A JP 2010527295A JP 5535921 B2 JP5535921 B2 JP 5535921B2
- Authority
- JP
- Japan
- Prior art keywords
- primer
- oligonucleotide
- sequence
- facilitator
- nucleic acid
- 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
- 230000003321 amplification Effects 0.000 title claims description 133
- 238000003199 nucleic acid amplification method Methods 0.000 title claims description 133
- 150000007523 nucleic acids Chemical class 0.000 title claims description 100
- 108020004707 nucleic acids Proteins 0.000 title claims description 86
- 102000039446 nucleic acids Human genes 0.000 title claims description 86
- 239000013615 primer Substances 0.000 claims description 323
- 108091034117 Oligonucleotide Proteins 0.000 claims description 298
- 238000000034 method Methods 0.000 claims description 94
- 230000027455 binding Effects 0.000 claims description 87
- 239000003999 initiator Substances 0.000 claims description 86
- 230000001419 dependent effect Effects 0.000 claims description 76
- 125000003729 nucleotide group Chemical group 0.000 claims description 75
- 239000002773 nucleotide Substances 0.000 claims description 66
- 238000003786 synthesis reaction Methods 0.000 claims description 61
- 230000015572 biosynthetic process Effects 0.000 claims description 60
- 230000004048 modification Effects 0.000 claims description 59
- 238000012986 modification Methods 0.000 claims description 59
- 108010044843 Peptide Initiation Factors Proteins 0.000 claims description 54
- 102000005877 Peptide Initiation Factors Human genes 0.000 claims description 54
- 238000006243 chemical reaction Methods 0.000 claims description 52
- 230000002441 reversible effect Effects 0.000 claims description 41
- 230000000295 complement effect Effects 0.000 claims description 38
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 claims description 28
- 238000005382 thermal cycling Methods 0.000 claims description 20
- 239000003155 DNA primer Substances 0.000 claims description 17
- 208000035657 Abasia Diseases 0.000 claims description 16
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 14
- 230000002255 enzymatic effect Effects 0.000 claims description 14
- 230000037452 priming Effects 0.000 claims description 14
- 108090000623 proteins and genes Proteins 0.000 claims description 11
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 claims description 10
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 9
- 230000008929 regeneration Effects 0.000 claims description 9
- 238000011069 regeneration method Methods 0.000 claims description 9
- 238000007792 addition Methods 0.000 claims description 8
- 201000010099 disease Diseases 0.000 claims description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 230000011987 methylation Effects 0.000 claims description 7
- 238000007069 methylation reaction Methods 0.000 claims description 7
- 230000007067 DNA methylation Effects 0.000 claims description 6
- 238000010348 incorporation Methods 0.000 claims description 6
- 125000006850 spacer group Chemical group 0.000 claims description 6
- 102000006943 Uracil-DNA Glycosidase Human genes 0.000 claims description 5
- 108010072685 Uracil-DNA Glycosidase Proteins 0.000 claims description 5
- 239000012472 biological sample Substances 0.000 claims description 5
- 229940035893 uracil Drugs 0.000 claims description 5
- 108091093037 Peptide nucleic acid Proteins 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 4
- 239000005546 dideoxynucleotide Substances 0.000 claims description 4
- -1 nucleotide amines Chemical class 0.000 claims description 4
- 230000001737 promoting effect Effects 0.000 claims description 4
- 238000012217 deletion Methods 0.000 claims description 3
- 230000037430 deletion Effects 0.000 claims description 3
- 238000003745 diagnosis Methods 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 238000006467 substitution reaction Methods 0.000 claims description 3
- 230000008859 change Effects 0.000 claims description 2
- 230000008034 disappearance Effects 0.000 claims description 2
- 230000009467 reduction Effects 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims 1
- 230000009149 molecular binding Effects 0.000 claims 1
- 238000003752 polymerase chain reaction Methods 0.000 description 52
- 108020004414 DNA Proteins 0.000 description 33
- 239000000523 sample Substances 0.000 description 20
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 18
- 230000035772 mutation Effects 0.000 description 14
- 238000001514 detection method Methods 0.000 description 13
- 238000009396 hybridization Methods 0.000 description 12
- 108091093088 Amplicon Proteins 0.000 description 11
- 230000010076 replication Effects 0.000 description 9
- 101000834948 Homo sapiens Tomoregulin-2 Proteins 0.000 description 8
- 102100026160 Tomoregulin-2 Human genes 0.000 description 8
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 8
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 8
- 238000000137 annealing Methods 0.000 description 7
- 239000008280 blood Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 238000010438 heat treatment Methods 0.000 description 6
- 238000003753 real-time PCR Methods 0.000 description 6
- 101000740968 Homo sapiens Transcription factor IIIB 50 kDa subunit Proteins 0.000 description 5
- 101000802101 Homo sapiens mRNA decay activator protein ZFP36L2 Proteins 0.000 description 5
- 102100039038 Transcription factor IIIB 50 kDa subunit Human genes 0.000 description 5
- 238000004925 denaturation Methods 0.000 description 5
- 230000036425 denaturation Effects 0.000 description 5
- 238000013461 design Methods 0.000 description 5
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 108091029523 CpG island Proteins 0.000 description 4
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 4
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 4
- 229930010555 Inosine Natural products 0.000 description 4
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 230000001351 cycling effect Effects 0.000 description 4
- 229940104302 cytosine Drugs 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 4
- 229960003786 inosine Drugs 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 229910052697 platinum Inorganic materials 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- OCLZPNCLRLDXJC-NTSWFWBYSA-N 2-amino-9-[(2r,5s)-5-(hydroxymethyl)oxolan-2-yl]-3h-purin-6-one Chemical compound C1=2NC(N)=NC(=O)C=2N=CN1[C@H]1CC[C@@H](CO)O1 OCLZPNCLRLDXJC-NTSWFWBYSA-N 0.000 description 3
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical compound CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 description 3
- 108010017826 DNA Polymerase I Proteins 0.000 description 3
- 102000004594 DNA Polymerase I Human genes 0.000 description 3
- 102100035189 GPI ethanolamine phosphate transferase 1 Human genes 0.000 description 3
- 101001093751 Homo sapiens GPI ethanolamine phosphate transferase 1 Proteins 0.000 description 3
- 101000984753 Homo sapiens Serine/threonine-protein kinase B-raf Proteins 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 3
- 102100027103 Serine/threonine-protein kinase B-raf Human genes 0.000 description 3
- 108010006785 Taq Polymerase Proteins 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 238000006911 enzymatic reaction Methods 0.000 description 3
- 239000007850 fluorescent dye Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000005457 optimization Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- SXUXMRMBWZCMEN-UHFFFAOYSA-N 2'-O-methyl uridine Natural products COC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 SXUXMRMBWZCMEN-UHFFFAOYSA-N 0.000 description 2
- SXUXMRMBWZCMEN-ZOQUXTDFSA-N 2'-O-methyluridine Chemical compound CO[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 SXUXMRMBWZCMEN-ZOQUXTDFSA-N 0.000 description 2
- 108091023043 Alu Element Proteins 0.000 description 2
- 108020004635 Complementary DNA Proteins 0.000 description 2
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 description 2
- 102100029764 DNA-directed DNA/RNA polymerase mu Human genes 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 2
- 241000713862 Moloney murine sarcoma virus Species 0.000 description 2
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 2
- 108700020796 Oncogene Proteins 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- 101100495925 Schizosaccharomyces pombe (strain 972 / ATCC 24843) chr3 gene Proteins 0.000 description 2
- 238000007844 allele-specific PCR Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 101150048834 braF gene Proteins 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 239000005547 deoxyribonucleotide Substances 0.000 description 2
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 238000007855 methylation-specific PCR Methods 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 238000007857 nested PCR Methods 0.000 description 2
- 239000002751 oligonucleotide probe Substances 0.000 description 2
- 238000002515 oligonucleotide synthesis Methods 0.000 description 2
- 231100000590 oncogenic Toxicity 0.000 description 2
- 230000002246 oncogenic effect Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000003362 replicative effect Effects 0.000 description 2
- 239000002336 ribonucleotide Substances 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 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 2
- HWPZZUQOWRWFDB-UHFFFAOYSA-N 1-methylcytosine Chemical compound CN1C=CC(N)=NC1=O HWPZZUQOWRWFDB-UHFFFAOYSA-N 0.000 description 1
- MXHRCPNRJAMMIM-SHYZEUOFSA-N 2'-deoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 MXHRCPNRJAMMIM-SHYZEUOFSA-N 0.000 description 1
- COCMHKNAGZHBDZ-UHFFFAOYSA-N 4-carboxy-3-[3-(dimethylamino)-6-dimethylazaniumylidenexanthen-9-yl]benzoate Chemical compound C=12C=CC(=[N+](C)C)C=C2OC2=CC(N(C)C)=CC=C2C=1C1=CC(C([O-])=O)=CC=C1C(O)=O COCMHKNAGZHBDZ-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 108091029430 CpG site Proteins 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 1
- AHCYMLUZIRLXAA-SHYZEUOFSA-N Deoxyuridine 5'-triphosphate Chemical compound O1[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C[C@@H]1N1C(=O)NC(=O)C=C1 AHCYMLUZIRLXAA-SHYZEUOFSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 208000026350 Inborn Genetic disease Diseases 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102000055027 Protein Methyltransferases Human genes 0.000 description 1
- 108700040121 Protein Methyltransferases Proteins 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical class OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- 241000205180 Thermococcus litoralis Species 0.000 description 1
- 101150015532 Tmeff2 gene Proteins 0.000 description 1
- 102000004357 Transferases Human genes 0.000 description 1
- 108090000992 Transferases Proteins 0.000 description 1
- 102100037111 Uracil-DNA glycosylase Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- HDRRAMINWIWTNU-PRJDIBJQSA-N [[(5r)-5-(2-amino-6-oxo-3h-purin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound C1=2NC(N)=NC(=O)C=2N=CN1[C@H]1CCC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HDRRAMINWIWTNU-PRJDIBJQSA-N 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 238000003491 array Methods 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 1
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 1
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 1
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 1
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 1
- MXHRCPNRJAMMIM-UHFFFAOYSA-N desoxyuridine Natural products C1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 MXHRCPNRJAMMIM-UHFFFAOYSA-N 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 208000016361 genetic disease Diseases 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000007403 mPCR Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 229920002113 octoxynol Polymers 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 239000012521 purified sample Substances 0.000 description 1
- 108700042226 ras Genes Proteins 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000012772 sequence design Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000012409 standard PCR amplification Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2007905445A AU2007905445A0 (en) | 2007-10-04 | Nucleic acid amplification | |
| AU2007905445 | 2007-10-04 | ||
| PCT/AU2008/001475 WO2009043112A1 (en) | 2007-10-04 | 2008-10-03 | Nucleic acid amplification |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2010539956A JP2010539956A (ja) | 2010-12-24 |
| JP2010539956A5 JP2010539956A5 (cg-RX-API-DMAC7.html) | 2011-11-17 |
| JP5535921B2 true JP5535921B2 (ja) | 2014-07-02 |
Family
ID=40525777
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2010527295A Active JP5535921B2 (ja) | 2007-10-04 | 2008-10-03 | 核酸増幅 |
Country Status (7)
| Country | Link |
|---|---|
| US (1) | US8455197B2 (cg-RX-API-DMAC7.html) |
| EP (1) | EP2207896B1 (cg-RX-API-DMAC7.html) |
| JP (1) | JP5535921B2 (cg-RX-API-DMAC7.html) |
| CN (1) | CN101889096B (cg-RX-API-DMAC7.html) |
| AU (1) | AU2008307153B2 (cg-RX-API-DMAC7.html) |
| ES (1) | ES2460896T3 (cg-RX-API-DMAC7.html) |
| WO (1) | WO2009043112A1 (cg-RX-API-DMAC7.html) |
Families Citing this family (33)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2271772B1 (en) * | 2008-03-11 | 2014-07-16 | Sequenom, Inc. | Nucleic acid-based tests for prenatal gender determination |
| US8962247B2 (en) | 2008-09-16 | 2015-02-24 | Sequenom, Inc. | Processes and compositions for methylation-based enrichment of fetal nucleic acid from a maternal sample useful for non invasive prenatal diagnoses |
| US8476013B2 (en) * | 2008-09-16 | 2013-07-02 | Sequenom, Inc. | Processes and compositions for methylation-based acid enrichment of fetal nucleic acid from a maternal sample useful for non-invasive prenatal diagnoses |
| WO2011009073A1 (en) * | 2009-07-16 | 2011-01-20 | Abbott Laboratories | A method for amplification of nucleic acids |
| DK2516680T3 (en) | 2009-12-22 | 2016-05-02 | Sequenom Inc | Method and kits to identify aneuploidy |
| EP2702168B1 (en) | 2011-04-29 | 2018-01-17 | Sequenom, Inc. | Quantification of a minority nucleic acid species |
| AU2012340118A1 (en) * | 2011-11-17 | 2014-04-24 | Rheonix, Inc. | System and methods for selective molecular analysis |
| US9605313B2 (en) | 2012-03-02 | 2017-03-28 | Sequenom, Inc. | Methods and processes for non-invasive assessment of genetic variations |
| DK3594366T3 (da) | 2012-05-11 | 2021-09-13 | Clinical Genomics Pty Ltd | Diagnostisk genmarkørpanel |
| US9920361B2 (en) | 2012-05-21 | 2018-03-20 | Sequenom, Inc. | Methods and compositions for analyzing nucleic acid |
| US10504613B2 (en) | 2012-12-20 | 2019-12-10 | Sequenom, Inc. | Methods and processes for non-invasive assessment of genetic variations |
| HK1206792A1 (en) | 2012-07-13 | 2016-01-15 | Sequenom, Inc. | Processes and compositions for methylation-based enrichment of fetal nucleic acid from a maternal sample useful for non-invasive prenatal diagnoses |
| CN104822840A (zh) * | 2012-12-13 | 2015-08-05 | 合成基因组股份有限公司 | Peg-介导的核酸分子的组装 |
| EP3597774A1 (en) | 2013-03-13 | 2020-01-22 | Sequenom, Inc. | Primers for dna methylation analysis |
| WO2014145176A1 (en) * | 2013-03-15 | 2014-09-18 | Arnold Lyle J | Methods for amplifying fragmented target nucleic acids utilizing an assembler sequence |
| CN113832216A (zh) * | 2013-03-15 | 2021-12-24 | 莱尔·J·阿诺德 | 使用发卡寡核苷酸扩增核酸的方法 |
| EP4321867B1 (en) * | 2013-08-09 | 2025-10-01 | Luminex Corporation | Probes for improved melt discrimination and multiplexing in nucleic acid assays |
| WO2015138774A1 (en) | 2014-03-13 | 2015-09-17 | Sequenom, Inc. | Methods and processes for non-invasive assessment of genetic variations |
| ES2861353T3 (es) * | 2014-05-19 | 2021-10-06 | Univ Rice William M | Amplificación específica de alelo utilizando una composición de oligonucleótidos cebadores no específicos de alelo y bloqueantes específicos de alelo solapantes |
| CN104131070A (zh) * | 2014-05-22 | 2014-11-05 | 苏州工业园区为真生物医药科技有限公司 | 一种对基因甲基化检测的方法 |
| EP3209801B1 (en) | 2014-10-20 | 2023-06-28 | Commonwealth Scientific and Industrial Research Organisation | Genome methylation analysis |
| CN105331727B (zh) * | 2015-12-01 | 2019-09-03 | 湖南宏雅基因技术有限公司 | 一种人外周血循环肿瘤DNA中septin 9基因甲基化的检测试剂盒 |
| GB201612011D0 (en) | 2016-07-11 | 2016-08-24 | Glaxosmithkline Ip Dev Ltd | Novel processes for the production of oligonucleotides |
| WO2018024562A1 (en) * | 2016-08-02 | 2018-02-08 | Roche Diagnostics Gmbh | Helper oligonucleotide for improving efficiency of amplification and detection/quantitation of nucleic acids |
| JP7001678B2 (ja) | 2016-09-06 | 2022-02-04 | スウィフト バイオサイエンシーズ， インコーポレイテッド | Ｎｇｓライブラリー濃度の正規化 |
| DE102016120124B8 (de) * | 2016-10-21 | 2018-08-23 | Gna Biosolutions Gmbh | Verfahren zum Durchführen einer Polymerase-Kettenreaktion und Vorrichtung zum Ausführen des Verfahrens |
| US10787699B2 (en) * | 2017-02-08 | 2020-09-29 | Microsoft Technology Licensing, Llc | Generating pluralities of primer and payload designs for retrieval of stored nucleotides |
| WO2019162529A1 (de) * | 2018-02-26 | 2019-08-29 | Agct Gmbh | Verfahren zur primer-verlängerungsreaktion mit verbesserter spezifität |
| EP3530753A1 (de) * | 2018-02-26 | 2019-08-28 | AGCT GmbH | Verfahren zur primer-verlängerungsreaktion mit verbesserter spezifität |
| EP3530754A1 (de) * | 2018-02-26 | 2019-08-28 | AGCT GmbH | Verfahren zur amplifikation einer nukleinsäure mit verbesserter spezifität |
| CN112074612A (zh) * | 2018-02-26 | 2020-12-11 | Agct有限公司 | 一种具有更高特异性的核酸扩增方法 |
| US20210071244A1 (en) * | 2018-02-28 | 2021-03-11 | Agct Gmbh | Method for the amplification of a nucleic acid with improved specificity |
| CN114075595B (zh) * | 2021-11-22 | 2023-11-17 | 上海交通大学 | 甲基化检测组合物、试剂盒及方法 |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6335184B1 (en) * | 1993-07-23 | 2002-01-01 | Bio-Rad Laboratories, Inc. | Linked linear amplification of nucleic acids |
| US6057134A (en) * | 1996-10-07 | 2000-05-02 | Ambion, Inc. | Modulating the efficiency of nucleic acid amplification reactions with 3' modified oligonucleotides |
| EP0866071B1 (en) * | 1997-03-20 | 2004-10-20 | F. Hoffmann-La Roche Ag | Modified primers |
| US7205129B1 (en) * | 2000-02-28 | 2007-04-17 | Qiagen Gmbh | Method for reducing artifacts in nucleic acid amplification |
| KR20040028991A (ko) * | 2001-08-20 | 2004-04-03 | 다카라 바이오 가부시키가이샤 | 핵산 증폭 방법 |
| EP1481088B1 (en) | 2002-03-01 | 2008-09-17 | Integrated Dna Technologies, Inc. | Polynomial amplification of nucleic acids |
| US7629152B2 (en) * | 2002-03-01 | 2009-12-08 | Integrated Dna Technologies, Inc. | Methods for amplifying polymeric nucleic acids |
| US20050153333A1 (en) | 2003-12-02 | 2005-07-14 | Sooknanan Roy R. | Selective terminal tagging of nucleic acids |
| US20060014183A1 (en) * | 2004-06-10 | 2006-01-19 | Pfundheller Henrik M | Extendable probes |
| US8501403B2 (en) * | 2006-03-28 | 2013-08-06 | Commonwealth Scientific And Industrial Research Organisation | Amplification of DNA fragments |
| WO2008064687A1 (en) * | 2006-11-27 | 2008-06-05 | Fluimedix | Fidelity enhanced allele specific amplification method |
-
2008
- 2008-10-03 CN CN200880119335.8A patent/CN101889096B/zh not_active Expired - Fee Related
- 2008-10-03 ES ES08800110.2T patent/ES2460896T3/es active Active
- 2008-10-03 WO PCT/AU2008/001475 patent/WO2009043112A1/en not_active Ceased
- 2008-10-03 JP JP2010527295A patent/JP5535921B2/ja active Active
- 2008-10-03 EP EP08800110.2A patent/EP2207896B1/en active Active
- 2008-10-03 AU AU2008307153A patent/AU2008307153B2/en not_active Ceased
- 2008-10-03 US US12/681,603 patent/US8455197B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| EP2207896A1 (en) | 2010-07-21 |
| WO2009043112A1 (en) | 2009-04-09 |
| EP2207896B1 (en) | 2014-01-29 |
| JP2010539956A (ja) | 2010-12-24 |
| CN101889096B (zh) | 2015-10-21 |
| ES2460896T3 (es) | 2014-05-14 |
| CN101889096A (zh) | 2010-11-17 |
| AU2008307153B2 (en) | 2015-03-12 |
| US20110033851A1 (en) | 2011-02-10 |
| EP2207896A4 (en) | 2010-11-10 |
| AU2008307153A1 (en) | 2009-04-09 |
| US8455197B2 (en) | 2013-06-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5535921B2 (ja) | 核酸増幅 | |
| CN107849603B (zh) | 利用有限核苷酸组成的引物扩增 | |
| AU2016281718B2 (en) | Selective degradation of wild-type DNA and enrichment of mutant alleles using nuclease | |
| US9518292B2 (en) | Methods for suppression PCR | |
| JP4989004B2 (ja) | 加ピロリン酸分解活性化重合（ｐａｐ）：アリル−特異的増幅および核酸配列決定への適応 | |
| EP2971172B1 (en) | Method for relative quantification of changes in dna methylation, using combined nuclease, ligation, and polymerase reactions | |
| JP5336350B2 (ja) | Ｄｎａフラグメントの増幅 | |
| JP6914831B2 (ja) | 標的核酸の高感度検出方法 | |
| EP2615171A1 (en) | Method for inhibiting nucleic acid amplification using light and highly sensitive method for selective nucleic acid amplification | |
| CN113227398A (zh) | 在减少的扩增时间下使用巢式重叠引物的指数基数-3核酸扩增 | |
| EP2982762B1 (en) | Nucleic acid amplification method using allele-specific reactive primer | |
| JP2007530026A (ja) | 核酸配列決定 | |
| CN114717298A (zh) | 一种多重扩增方法 | |
| HK40064470A (en) | Amplification with primers of limited nucleotide composition | |
| WO2018009677A1 (en) | Fast target enrichment by multiplexed relay pcr with modified bubble primers | |
| JP2009219445A (ja) | Ｖｋｏｒｃ１遺伝子上の１塩基多型を検出する方法およびプライマーセット | |
| HK40000246A (en) | Nucleic acid amplification method using allele-specific reactive primer | |
| HK40000246B (en) | Nucleic acid amplification method using allele-specific reactive primer | |
| Whitcombe | 6 Using Scorpion Primers for Genotyping | |
| HK1184814A (en) | Inhibition method of nucleic acid amplification by photoirradiation and method of selective nucleic acid amplification with high sensitivity | |
| HK1162195A (en) | Methods, compositions, and kits for detecting allelic variants |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110930 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20110930 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20130827 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20131126 |
|
| A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20131203 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20140124 |
|
| A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20140131 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20140227 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20140324 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20140423 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 5535921 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |