JP2005517455A - 核酸定量化 - Google Patents
核酸定量化 Download PDFInfo
- Publication number
- JP2005517455A JP2005517455A JP2003569871A JP2003569871A JP2005517455A JP 2005517455 A JP2005517455 A JP 2005517455A JP 2003569871 A JP2003569871 A JP 2003569871A JP 2003569871 A JP2003569871 A JP 2003569871A JP 2005517455 A JP2005517455 A JP 2005517455A
- Authority
- JP
- Japan
- Prior art keywords
- real
- nucleic acid
- time pcr
- quantification
- cdna
- 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.)
- Pending
Links
- 238000001921 nucleic acid quantification Methods 0.000 title claims description 10
- 238000000034 method Methods 0.000 claims abstract description 79
- 239000002299 complementary DNA Substances 0.000 claims abstract description 61
- 238000011002 quantification Methods 0.000 claims abstract description 56
- 238000003753 real-time PCR Methods 0.000 claims abstract description 51
- 239000013612 plasmid Substances 0.000 claims abstract description 40
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 36
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 27
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 27
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 27
- 239000000523 sample Substances 0.000 claims description 42
- 108020004414 DNA Proteins 0.000 claims description 33
- 102100037850 Interferon gamma Human genes 0.000 claims description 29
- 108010074328 Interferon-gamma Proteins 0.000 claims description 29
- 102000004127 Cytokines Human genes 0.000 claims description 27
- 108090000695 Cytokines Proteins 0.000 claims description 27
- ZYFVNVRFVHJEIU-UHFFFAOYSA-N PicoGreen Chemical compound CN(C)CCCN(CCCN(C)C)C1=CC(=CC2=[N+](C3=CC=CC=C3S2)C)C2=CC=CC=C2N1C1=CC=CC=C1 ZYFVNVRFVHJEIU-UHFFFAOYSA-N 0.000 claims description 22
- 108091093088 Amplicon Proteins 0.000 claims description 16
- 239000003153 chemical reaction reagent Substances 0.000 claims description 15
- 108010002350 Interleukin-2 Proteins 0.000 claims description 13
- 102000000588 Interleukin-2 Human genes 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 108090000174 Interleukin-10 Proteins 0.000 claims description 10
- 102000003814 Interleukin-10 Human genes 0.000 claims description 10
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 9
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 9
- 102000004169 proteins and genes Human genes 0.000 claims description 9
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims description 8
- 230000035945 sensitivity Effects 0.000 claims description 8
- 102000053602 DNA Human genes 0.000 claims description 7
- 238000003556 assay Methods 0.000 claims description 6
- 238000012360 testing method Methods 0.000 abstract description 16
- 230000008859 change Effects 0.000 abstract description 5
- 239000013615 primer Substances 0.000 description 24
- 238000003752 polymerase chain reaction Methods 0.000 description 23
- 210000004027 cell Anatomy 0.000 description 21
- 108020004999 messenger RNA Proteins 0.000 description 19
- 102000004388 Interleukin-4 Human genes 0.000 description 14
- 108090000978 Interleukin-4 Proteins 0.000 description 14
- 230000014509 gene expression Effects 0.000 description 14
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 14
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 14
- 238000013382 DNA quantification Methods 0.000 description 13
- 238000010790 dilution Methods 0.000 description 13
- 239000012895 dilution Substances 0.000 description 13
- 229940028885 interleukin-4 Drugs 0.000 description 13
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 11
- 238000003757 reverse transcription PCR Methods 0.000 description 11
- 239000002609 medium Substances 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 9
- 229940076144 interleukin-10 Drugs 0.000 description 9
- PHEDXBVPIONUQT-UHFFFAOYSA-N Cocarcinogen A1 Natural products CCCCCCCCCCCCCC(=O)OC1C(C)C2(O)C3C=C(C)C(=O)C3(O)CC(CO)=CC2C2C1(OC(C)=O)C2(C)C PHEDXBVPIONUQT-UHFFFAOYSA-N 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 238000004364 calculation method Methods 0.000 description 8
- 229960003130 interferon gamma Drugs 0.000 description 8
- PHEDXBVPIONUQT-RGYGYFBISA-N phorbol 13-acetate 12-myristate Chemical compound C([C@]1(O)C(=O)C(C)=C[C@H]1[C@@]1(O)[C@H](C)[C@H]2OC(=O)CCCCCCCCCCCCC)C(CO)=C[C@H]1[C@H]1[C@]2(OC(C)=O)C1(C)C PHEDXBVPIONUQT-RGYGYFBISA-N 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 239000002158 endotoxin Substances 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 7
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 description 7
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 description 7
- 229920006008 lipopolysaccharide Polymers 0.000 description 7
- 210000003705 ribosome Anatomy 0.000 description 7
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 6
- 239000002953 phosphate buffered saline Substances 0.000 description 6
- 210000004989 spleen cell Anatomy 0.000 description 6
- 239000011324 bead Substances 0.000 description 5
- 238000010804 cDNA synthesis Methods 0.000 description 5
- 238000010367 cloning Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- VLEIUWBSEKKKFX-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid Chemical compound OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O VLEIUWBSEKKKFX-UHFFFAOYSA-N 0.000 description 4
- 108091034117 Oligonucleotide Proteins 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000003068 molecular probe Substances 0.000 description 4
- 108020004418 ribosomal RNA Proteins 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 108700039887 Essential Genes Proteins 0.000 description 3
- 108060002716 Exonuclease Proteins 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 238000002123 RNA extraction Methods 0.000 description 3
- 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 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 102000013165 exonuclease Human genes 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 210000004698 lymphocyte Anatomy 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 230000001915 proofreading effect Effects 0.000 description 3
- 210000000952 spleen Anatomy 0.000 description 3
- 102000016911 Deoxyribonucleases Human genes 0.000 description 2
- 108010053770 Deoxyribonucleases Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 210000004976 peripheral blood cell Anatomy 0.000 description 2
- 239000002987 primer (paints) Substances 0.000 description 2
- 238000013207 serial dilution Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000013595 supernatant sample Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 108700031361 Brachyury Proteins 0.000 description 1
- 201000006474 Brain Ischemia Diseases 0.000 description 1
- 238000009631 Broth culture Methods 0.000 description 1
- 206010008120 Cerebral ischaemia Diseases 0.000 description 1
- 108020004638 Circular DNA Proteins 0.000 description 1
- 108020004635 Complementary DNA Proteins 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 102000008214 Glutamate decarboxylase Human genes 0.000 description 1
- 108091022930 Glutamate decarboxylase Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 102000019223 Interleukin-1 receptor Human genes 0.000 description 1
- 108050006617 Interleukin-1 receptor Proteins 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 239000006137 Luria-Bertani broth Substances 0.000 description 1
- 241000713869 Moloney murine leukemia virus Species 0.000 description 1
- 101001044384 Mus musculus Interferon gamma Proteins 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 244000025272 Persea americana Species 0.000 description 1
- 235000008673 Persea americana Nutrition 0.000 description 1
- 238000012341 Quantitative reverse-transcriptase PCR Methods 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 101710141795 Ribonuclease inhibitor Proteins 0.000 description 1
- 229940122208 Ribonuclease inhibitor Drugs 0.000 description 1
- 102100037968 Ribonuclease inhibitor Human genes 0.000 description 1
- CGNLCCVKSWNSDG-UHFFFAOYSA-N SYBR Green I Chemical compound CN(C)CCCN(CCC)C1=CC(C=C2N(C3=CC=CC=C3S2)C)=C2C=CC=CC2=[N+]1C1=CC=CC=C1 CGNLCCVKSWNSDG-UHFFFAOYSA-N 0.000 description 1
- 238000010818 SYBR green PCR Master Mix Methods 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 241000589500 Thermus aquaticus Species 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 230000003092 anti-cytokine Effects 0.000 description 1
- 238000003491 array Methods 0.000 description 1
- 230000003305 autocrine Effects 0.000 description 1
- 239000003710 calcium ionophore Substances 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 230000003822 cell turnover Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 206010008118 cerebral infarction Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 239000011888 foil Substances 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 210000005104 human peripheral blood lymphocyte Anatomy 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004968 inflammatory condition Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000010841 mRNA extraction Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000003226 mitogen Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 238000007899 nucleic acid hybridization Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 210000003024 peritoneal macrophage Anatomy 0.000 description 1
- 238000006303 photolysis reaction Methods 0.000 description 1
- 230000015843 photosynthesis, light reaction Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 239000003161 ribonuclease inhibitor Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
Images
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
- C12Q1/6851—Quantitative amplification
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (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 (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GBGB0204022.8A GB0204022D0 (en) | 2002-02-20 | 2002-02-20 | Nucleic acid quantification |
| PCT/GB2003/000079 WO2003070980A2 (en) | 2002-02-20 | 2003-01-10 | Nucleic acid quantification |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2005517455A true JP2005517455A (ja) | 2005-06-16 |
| JP2005517455A5 JP2005517455A5 (enExample) | 2006-03-02 |
Family
ID=9931449
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2003569871A Pending JP2005517455A (ja) | 2002-02-20 | 2003-01-10 | 核酸定量化 |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US20050158722A1 (enExample) |
| EP (1) | EP1495136B1 (enExample) |
| JP (1) | JP2005517455A (enExample) |
| AT (1) | ATE347620T1 (enExample) |
| AU (1) | AU2003206993A1 (enExample) |
| DE (1) | DE60310217D1 (enExample) |
| GB (1) | GB0204022D0 (enExample) |
| WO (1) | WO2003070980A2 (enExample) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2015192666A (ja) * | 2014-03-19 | 2015-11-05 | ハウス食品グループ本社株式会社 | リアルタイムｐｃｒによる食物アレルゲンの検出方法及びキット |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0426188D0 (en) * | 2004-11-30 | 2004-12-29 | Leuven K U Res & Dev | DNA repeats |
| FR2904833A1 (fr) * | 2006-08-11 | 2008-02-15 | Bioquanta Sarl | Procede de dosage d'acide nuclieque par fluorescence |
| US8346485B2 (en) | 2008-11-25 | 2013-01-01 | Quest Diagnostics Investments Incorporated | Methods and apparatuses for estimating initial target nucleic acid concentration in a sample by modeling background signal and cycle-dependent amplification efficiency of a polymerase chain reaction |
| CN102363814B (zh) * | 2011-11-23 | 2013-08-28 | 山西农业大学 | 一种绝对荧光定量pcr检测方法 |
| GB201720088D0 (en) | 2017-12-01 | 2018-01-17 | Univ Oslo Hf | PCR Controls |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5219727A (en) * | 1989-08-21 | 1993-06-15 | Hoffmann-Laroche Inc. | Quantitation of nucleic acids using the polymerase chain reaction |
| US6691041B2 (en) * | 2000-03-31 | 2004-02-10 | Roche Molecular Systems, Inc. | Method for the efficiency-corrected real-time quantification of nucleic acids |
-
2002
- 2002-02-20 GB GBGB0204022.8A patent/GB0204022D0/en not_active Ceased
-
2003
- 2003-01-10 DE DE60310217T patent/DE60310217D1/de not_active Expired - Lifetime
- 2003-01-10 WO PCT/GB2003/000079 patent/WO2003070980A2/en not_active Ceased
- 2003-01-10 AU AU2003206993A patent/AU2003206993A1/en not_active Abandoned
- 2003-01-10 US US10/503,596 patent/US20050158722A1/en not_active Abandoned
- 2003-01-10 JP JP2003569871A patent/JP2005517455A/ja active Pending
- 2003-01-10 AT AT03704733T patent/ATE347620T1/de not_active IP Right Cessation
- 2003-01-10 EP EP03704733A patent/EP1495136B1/en not_active Expired - Lifetime
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2015192666A (ja) * | 2014-03-19 | 2015-11-05 | ハウス食品グループ本社株式会社 | リアルタイムｐｃｒによる食物アレルゲンの検出方法及びキット |
Also Published As
| Publication number | Publication date |
|---|---|
| AU2003206993A1 (en) | 2003-09-09 |
| US20050158722A1 (en) | 2005-07-21 |
| ATE347620T1 (de) | 2006-12-15 |
| EP1495136B1 (en) | 2006-12-06 |
| GB0204022D0 (en) | 2002-04-03 |
| WO2003070980A2 (en) | 2003-08-28 |
| DE60310217D1 (de) | 2007-01-18 |
| EP1495136A2 (en) | 2005-01-12 |
| WO2003070980A3 (en) | 2003-12-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Whelan et al. | A method for the absolute quantification of cDNA using real-time PCR | |
| Giulietti et al. | An overview of real-time quantitative PCR: applications to quantify cytokine gene expression | |
| Buga et al. | Identification of new therapeutic targets by genome-wide analysis of gene expression in the ipsilateral cortex of aged rats after stroke | |
| Leutenegger et al. | Quantitative real-time PCR for the measurement of feline cytokine mRNA | |
| Kruse et al. | Quantification of cytokine mRNA expression by RT PCR in samples of previously frozen blood | |
| Babu et al. | Limitations and modifications of quantitative polymerase chain reaction: application to measurement of multiple mRNAs present in small amounts of sample RNA | |
| Vernel-Pauillac et al. | Proinflammatory and Immunomodulatory Cytokine mRNA Time Course Profiles in Hamsters Infected with a Virulent Variant of Leptospirainterrogans | |
| US20030087259A1 (en) | Methods and compositions for regulating bone and cartilage formation | |
| EP2999800B1 (en) | Measures of short telomere abundance | |
| Ausiello et al. | Cytokine gene expression in human peripheral blood mononuclear cells stimulated by mannoprotein constituents from Candida albicans | |
| Maeda et al. | Lesional expression of thymus and activation-regulated chemokine in canine atopic dermatitis | |
| Haugland et al. | Expression profiles of inflammatory and immune-related genes in Atlantic salmon (Salmo salar L.) at early time post vaccination | |
| US20080200381A1 (en) | Methods and compositions for regulating bone and cartilage formation | |
| Plotnikova et al. | Development of a multiplex quantitative PCR assay for the analysis of human cytokine gene expression in influenza A virus-infected cells | |
| JP2005517455A (ja) | 核酸定量化 | |
| CN114350849A (zh) | 检测SARS-CoV-2的组合物、试剂盒、方法及其用途 | |
| Shibata et al. | Quantitative polymerase chain reaction using an external control mRNA for determination of gene expression in a heterogeneous cell population. | |
| Muise et al. | Comparison of inhibition of ovalbumin-induced bronchoconstriction in guinea pigs and in vitro inhibition of tumor necrosis factor-α formation with phosphodiesterase 4 (PDE4) selective inhibitors | |
| Morozumi et al. | Elevated mRNA expression for supervillin and vascular endothelial growth factor in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis | |
| US20230417747A1 (en) | Methods and compositions for diagnosing and treating virally-associated disease | |
| Dutta et al. | Immune response during influenza virus infection among the population of Assam, Northeast India | |
| Salazar-Montes et al. | Chemically induced liver regeneration is characterized by specific IL-6 gene expression | |
| Della Pietra et al. | Highly conserved amino-acid sequence between murine STAT3 and a revised human STAT3 sequence | |
| Menne et al. | Real-time polymerase chain reaction assays for leukocyte CD and cytokine mRNAs of the Eastern woodchuck (Marmota monax) | |
| Koning et al. | Analysis of cyt0kine gene expression in stimulated T cells of small children by semi‐quantitative PCR |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20060110 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20060110 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20090127 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20090630 |