JP5331404B2 - 先天性異常症の染色体欠失の検出方法 - Google Patents
先天性異常症の染色体欠失の検出方法 Download PDFInfo
- Publication number
- JP5331404B2 JP5331404B2 JP2008199541A JP2008199541A JP5331404B2 JP 5331404 B2 JP5331404 B2 JP 5331404B2 JP 2008199541 A JP2008199541 A JP 2008199541A JP 2008199541 A JP2008199541 A JP 2008199541A JP 5331404 B2 JP5331404 B2 JP 5331404B2
- Authority
- JP
- Japan
- Prior art keywords
- region
- dna
- nucleic acid
- syndrome
- deletion
- 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
- 238000000034 method Methods 0.000 title claims description 72
- 206010061764 Chromosomal deletion Diseases 0.000 title claims description 8
- 206010010356 Congenital anomaly Diseases 0.000 title description 8
- 238000012217 deletion Methods 0.000 claims description 44
- 230000037430 deletion Effects 0.000 claims description 44
- 108020004707 nucleic acids Proteins 0.000 claims description 44
- 102000039446 nucleic acids Human genes 0.000 claims description 44
- 150000007523 nucleic acids Chemical class 0.000 claims description 44
- 210000000349 chromosome Anatomy 0.000 claims description 36
- 238000001514 detection method Methods 0.000 claims description 31
- 208000036626 Mental retardation Diseases 0.000 claims description 21
- 208000033787 Rare developmental defect during embryogenesis Diseases 0.000 claims description 19
- 208000022734 developmental defect during embryogenesis Diseases 0.000 claims description 19
- 208000035853 malformation syndrome Diseases 0.000 claims description 19
- 238000009396 hybridization Methods 0.000 claims description 18
- 239000000758 substrate Substances 0.000 claims description 18
- 210000003917 human chromosome Anatomy 0.000 claims description 12
- 108091034117 Oligonucleotide Proteins 0.000 claims description 10
- 238000002105 Southern blotting Methods 0.000 claims description 7
- 238000000636 Northern blotting Methods 0.000 claims description 6
- 238000003753 real-time PCR Methods 0.000 claims description 6
- 238000000018 DNA microarray Methods 0.000 claims description 4
- 239000002299 complementary DNA Substances 0.000 claims description 4
- 230000007812 deficiency Effects 0.000 claims 1
- 108020004414 DNA Proteins 0.000 description 73
- 210000004436 artificial bacterial chromosome Anatomy 0.000 description 25
- 239000000523 sample Substances 0.000 description 25
- 238000004458 analytical method Methods 0.000 description 24
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 22
- 208000011580 syndromic disease Diseases 0.000 description 22
- 201000010099 disease Diseases 0.000 description 15
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 14
- 238000000746 purification Methods 0.000 description 14
- 108090000623 proteins and genes Proteins 0.000 description 13
- 230000005856 abnormality Effects 0.000 description 11
- 239000012528 membrane Substances 0.000 description 9
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 8
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 8
- 230000003321 amplification Effects 0.000 description 8
- 238000003199 nucleic acid amplification method Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- 239000000377 silicon dioxide Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 6
- 208000035475 disorder Diseases 0.000 description 6
- 238000003745 diagnosis Methods 0.000 description 5
- 239000007850 fluorescent dye Substances 0.000 description 5
- 239000011521 glass Substances 0.000 description 5
- 230000031864 metaphase Effects 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 206010060860 Neurological symptom Diseases 0.000 description 4
- 230000002159 abnormal effect Effects 0.000 description 4
- 210000001106 artificial yeast chromosome Anatomy 0.000 description 4
- 229960002685 biotin Drugs 0.000 description 4
- 235000020958 biotin Nutrition 0.000 description 4
- 239000011616 biotin Substances 0.000 description 4
- 230000002759 chromosomal effect Effects 0.000 description 4
- 238000012869 ethanol precipitation Methods 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 238000002493 microarray Methods 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 230000004544 DNA amplification Effects 0.000 description 3
- 101000610557 Homo sapiens U4/U6 small nuclear ribonucleoprotein Prp31 Proteins 0.000 description 3
- 101001109965 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) 60S ribosomal protein L7-A Proteins 0.000 description 3
- 101001109960 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) 60S ribosomal protein L7-B Proteins 0.000 description 3
- 208000036623 Severe mental retardation Diseases 0.000 description 3
- 102100040118 U4/U6 small nuclear ribonucleoprotein Prp31 Human genes 0.000 description 3
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical compound C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000002795 fluorescence method Methods 0.000 description 3
- 229910010272 inorganic material Inorganic materials 0.000 description 3
- 210000004698 lymphocyte Anatomy 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 208000003278 patent ductus arteriosus Diseases 0.000 description 3
- 238000002205 phenol-chloroform extraction Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 108091005957 yellow fluorescent proteins Proteins 0.000 description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical group C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 2
- 208000010543 22q11.2 deletion syndrome Diseases 0.000 description 2
- 208000034431 Adrenal hypoplasia congenita Diseases 0.000 description 2
- 201000011374 Alagille syndrome Diseases 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 208000005875 Alternating hemiplegia of childhood Diseases 0.000 description 2
- 208000009575 Angelman syndrome Diseases 0.000 description 2
- 206010005155 Blepharophimosis Diseases 0.000 description 2
- 208000014392 Cat-eye syndrome Diseases 0.000 description 2
- 208000003449 Classical Lissencephalies and Subcortical Band Heterotopias Diseases 0.000 description 2
- 206010009260 Cleft lip and palate Diseases 0.000 description 2
- 206010010904 Convulsion Diseases 0.000 description 2
- 206010049889 Craniosynostosis Diseases 0.000 description 2
- 206010011385 Cri-du-chat syndrome Diseases 0.000 description 2
- -1 DAPI Chemical compound 0.000 description 2
- 208000000398 DiGeorge Syndrome Diseases 0.000 description 2
- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 description 2
- 201000010374 Down Syndrome Diseases 0.000 description 2
- 206010013801 Duchenne Muscular Dystrophy Diseases 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108010022894 Euchromatin Proteins 0.000 description 2
- 208000034454 F12-related hereditary angioedema with normal C1Inh Diseases 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 206010050638 Langer-Giedion syndrome Diseases 0.000 description 2
- 201000004246 Miller-Dieker lissencephaly syndrome Diseases 0.000 description 2
- 208000035022 Miller-Dieker syndrome Diseases 0.000 description 2
- 208000001804 Monosomy 5p Diseases 0.000 description 2
- 208000003019 Neurofibromatosis 1 Diseases 0.000 description 2
- 101100384865 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) cot-1 gene Proteins 0.000 description 2
- 208000017493 Pelizaeus-Merzbacher disease Diseases 0.000 description 2
- 206010039281 Rubinstein-Taybi syndrome Diseases 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 206010062282 Silver-Russell syndrome Diseases 0.000 description 2
- 201000001388 Smith-Magenis syndrome Diseases 0.000 description 2
- 201000003696 Sotos syndrome Diseases 0.000 description 2
- 208000004350 Strabismus Diseases 0.000 description 2
- 108091081400 Subtelomere Proteins 0.000 description 2
- 208000020741 Tetrasomy 12p Diseases 0.000 description 2
- 206010051956 Trichorhinophalangeal syndrome Diseases 0.000 description 2
- 208000035378 Trichorhinophalangeal syndrome type 2 Diseases 0.000 description 2
- 201000002919 Van der Woude syndrome Diseases 0.000 description 2
- 201000007960 WAGR syndrome Diseases 0.000 description 2
- 206010049644 Williams syndrome Diseases 0.000 description 2
- 235000010724 Wisteria floribunda Nutrition 0.000 description 2
- 208000006254 Wolf-Hirschhorn Syndrome Diseases 0.000 description 2
- 108091005948 blue fluorescent proteins Proteins 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 210000005252 bulbus oculi Anatomy 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000013611 chromosomal DNA Substances 0.000 description 2
- 208000016653 cleft lip/palate Diseases 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 description 2
- SHIBSTMRCDJXLN-KCZCNTNESA-N digoxigenin Chemical compound C1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)C[C@H]3O)C)=CC(=O)OC1 SHIBSTMRCDJXLN-KCZCNTNESA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 210000000632 euchromatin Anatomy 0.000 description 2
- 230000007717 exclusion Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 210000001508 eye Anatomy 0.000 description 2
- 230000001815 facial effect Effects 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 2
- 238000001215 fluorescent labelling Methods 0.000 description 2
- 230000005861 gene abnormality Effects 0.000 description 2
- 238000012224 gene deletion Methods 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 210000003128 head Anatomy 0.000 description 2
- 208000016861 hereditary angioedema type 3 Diseases 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 238000007901 in situ hybridization Methods 0.000 description 2
- 150000002484 inorganic compounds Chemical class 0.000 description 2
- 239000003456 ion exchange resin Substances 0.000 description 2
- 229920003303 ion-exchange polymer Polymers 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000013507 mapping Methods 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108010054624 red fluorescent protein Proteins 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 201000006532 trichorhinophalangeal syndrome type II Diseases 0.000 description 2
- 201000003130 ventricular septal defect Diseases 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- NNJPGOLRFBJNIW-HNNXBMFYSA-N (-)-demecolcine Chemical compound C1=C(OC)C(=O)C=C2[C@@H](NC)CCC3=CC(OC)=C(OC)C(OC)=C3C2=C1 NNJPGOLRFBJNIW-HNNXBMFYSA-N 0.000 description 1
- VXTQMOSXLCOXRL-UHFFFAOYSA-N (4-chloro-1h-indol-3-yl) dihydrogen phosphate Chemical compound C1=CC(Cl)=C2C(OP(O)(=O)O)=CNC2=C1 VXTQMOSXLCOXRL-UHFFFAOYSA-N 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 206010003497 Asphyxia Diseases 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 229910004613 CdTe Inorganic materials 0.000 description 1
- 229920002284 Cellulose triacetate Polymers 0.000 description 1
- 208000031404 Chromosome Aberrations Diseases 0.000 description 1
- 208000031639 Chromosome Deletion Diseases 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 208000001353 Coffin-Lowry syndrome Diseases 0.000 description 1
- 208000032170 Congenital Abnormalities Diseases 0.000 description 1
- 208000029767 Congenital, Hereditary, and Neonatal Diseases and Abnormalities Diseases 0.000 description 1
- 206010011878 Deafness Diseases 0.000 description 1
- NNJPGOLRFBJNIW-UHFFFAOYSA-N Demecolcine Natural products C1=C(OC)C(=O)C=C2C(NC)CCC3=CC(OC)=C(OC)C(OC)=C3C2=C1 NNJPGOLRFBJNIW-UHFFFAOYSA-N 0.000 description 1
- 208000001692 Esotropia Diseases 0.000 description 1
- 201000005538 Exotropia Diseases 0.000 description 1
- 206010015995 Eyelid ptosis Diseases 0.000 description 1
- 208000035478 Interatrial communication Diseases 0.000 description 1
- 206010050183 Macrocephaly Diseases 0.000 description 1
- 208000005767 Megalencephaly Diseases 0.000 description 1
- 208000009795 Microphthalmos Diseases 0.000 description 1
- 206010028182 Multiple congenital abnormalities Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 206010061534 Oesophageal squamous cell carcinoma Diseases 0.000 description 1
- 108010047620 Phytohemagglutinins Proteins 0.000 description 1
- 201000010769 Prader-Willi syndrome Diseases 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 208000036765 Squamous cell carcinoma of the esophagus Diseases 0.000 description 1
- 108010087999 Steryl-Sulfatase Proteins 0.000 description 1
- 102100038021 Steryl-sulfatase Human genes 0.000 description 1
- 208000037280 Trisomy Diseases 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 208000001910 Ventricular Heart Septal Defects Diseases 0.000 description 1
- NNLVGZFZQQXQNW-ADJNRHBOSA-N [(2r,3r,4s,5r,6s)-4,5-diacetyloxy-3-[(2s,3r,4s,5r,6r)-3,4,5-triacetyloxy-6-(acetyloxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6s)-4,5,6-triacetyloxy-2-(acetyloxymethyl)oxan-3-yl]oxyoxan-2-yl]methyl acetate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](OC(C)=O)[C@H]1OC(C)=O)O[C@H]1[C@@H]([C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H](COC(C)=O)O1)OC(C)=O)COC(=O)C)[C@@H]1[C@@H](COC(C)=O)O[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@H]1OC(C)=O NNLVGZFZQQXQNW-ADJNRHBOSA-N 0.000 description 1
- 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 1
- DPKHZNPWBDQZCN-UHFFFAOYSA-N acridine orange free base Chemical compound C1=CC(N(C)C)=CC2=NC3=CC(N(C)C)=CC=C3C=C21 DPKHZNPWBDQZCN-UHFFFAOYSA-N 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 210000004381 amniotic fluid Anatomy 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000003491 array Methods 0.000 description 1
- 210000004507 artificial chromosome Anatomy 0.000 description 1
- 208000013914 atrial heart septal defect Diseases 0.000 description 1
- 206010003664 atrial septal defect Diseases 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000002981 blocking agent Substances 0.000 description 1
- UHYPYGJEEGLRJD-UHFFFAOYSA-N cadmium(2+);selenium(2-) Chemical compound [Se-2].[Cd+2] UHYPYGJEEGLRJD-UHFFFAOYSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000006059 cover glass Substances 0.000 description 1
- 229940041984 dextran 1 Drugs 0.000 description 1
- 229960000633 dextran sulfate Drugs 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 208000007276 esophageal squamous cell carcinoma Diseases 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 210000004709 eyebrow Anatomy 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000010370 hearing loss Effects 0.000 description 1
- 231100000888 hearing loss Toxicity 0.000 description 1
- 208000016354 hearing loss disease Diseases 0.000 description 1
- 239000000815 hypotonic solution Substances 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 201000010478 microphthalmia Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- JPXMTWWFLBLUCD-UHFFFAOYSA-N nitro blue tetrazolium(2+) Chemical compound COC1=CC(C=2C=C(OC)C(=CC=2)[N+]=2N(N=C(N=2)C=2C=CC=CC=2)C=2C=CC(=CC=2)[N+]([O-])=O)=CC=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=C([N+]([O-])=O)C=C1 JPXMTWWFLBLUCD-UHFFFAOYSA-N 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 208000029278 non-syndromic brachydactyly of fingers Diseases 0.000 description 1
- 238000002966 oligonucleotide array Methods 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 208000024335 physical disease Diseases 0.000 description 1
- 230000001885 phytohemagglutinin Effects 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920000729 poly(L-lysine) polymer Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 201000003004 ptosis Diseases 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 229910052761 rare earth metal Inorganic materials 0.000 description 1
- 150000002910 rare earth metals Chemical class 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- SBIBMFFZSBJNJF-UHFFFAOYSA-N selenium;zinc Chemical compound [Se]=[Zn] SBIBMFFZSBJNJF-UHFFFAOYSA-N 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000009919 sequestration Effects 0.000 description 1
- 210000003765 sex chromosome Anatomy 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- FZHAPNGMFPVSLP-UHFFFAOYSA-N silanamine Chemical compound [SiH3]N FZHAPNGMFPVSLP-UHFFFAOYSA-N 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000001550 testis Anatomy 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
- 208000011908 tetrasomy Diseases 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
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 239000002966 varnish Substances 0.000 description 1
- 238000007631 vascular surgery Methods 0.000 description 1
- 210000003905 vulva Anatomy 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
- C12Q1/6876—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
-
- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6841—In situ hybridisation
-
- 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
- C12Q2600/156—Polymorphic or mutational markers
Description
(1)DNAアレイ
上述したように、本発明の検出方法を行うに際しては、オリゴヌクレオチド、cDNA、BAC DNA、PAC DNA、又は、YAC DNA等のプローブ核酸を搭載したDNAアレイを用いることが好適である。当該アレイとして用いられる基板の素材としては、ガラス、プラスチック、メンブレン、3次元アレイ等が用いられるが、スライドガラス等のガラス基板が一般的には好ましい。ガラス等の固体基板は、ポリ-L-リジン、アミノシラン、金・アルミニウム等の凝着により基板をコートされていることがより好ましい。
極めて有力な本発明の検出方法を行う態様として、染色体を網羅的にカバーするBAC DNAを搭載したMCG Whole Genome Array-4500(WGA−4500と略称することもある:非特許文献1)を用いたアレイCGH(Comparative Genomic Hybridization)法を行うことができる。WGA−4500は、22種類の常染色体とX、Y性染色体の全染色体を網羅する4523個のBACクローンを搭載した、平均解像度が約0.7Mbの全ゲノムアレイである。これはヒト染色体ユークロマチン領域の約1/3に及んでいる。WGA−4500を用いる場合、10q24.31-10q25.1の領域の一部を含むプローブ核酸の吸着量比の底が2の対数比が−0.3以下である場合を、本発明にかかわるヘミ接合体欠失の検出として認めることが通常であるが、この基準に限定されるものではない。
また、アレイに搭載するプローブ核酸を、上記のヘミ接合体欠失のパターンに合わせて絞り込むことで減少させた、DNA-アレイを用いることもできる。ただし、当該DNAアレイには少なくとも10q24.31-10q25.1の領域の一部を含むプローブ核酸を有していることが必要である。例えば、プローブ核酸としてBAC DNAを用いる場合、少なくとも、上記の22種のBAC DNA群から選択される1種以上のBAC DNAが固定化されていることが好適である。また、同時に、10q24.31-10q25.1領域以外に該当するBAC DNAが1種以上固定化されていることがより好適である。
DNAアレイの作成において、通常に得られるBAC DNA等は、ゲノムDNA定着基板を多数製造して実用化するには少量であるので、当該DNAを遺伝子増幅産物として得ることが好適である(この遺伝子増幅工程を「無尽蔵化」ともいう)。無尽蔵化においては、まずBAC DNA等を、4塩基認識酵素、例えば、RsaI、DpnI、HaeIII等で消化した後、アダプターを加えてライゲーションを行う。アダプターは10〜30塩基、好適には15〜25塩基からなるオリゴヌクレオチドで、2本鎖は相補的配列を有し、アニーリング後、平滑末端を形成する側の3’−末端のオリゴヌクレオチドをリン酸化する必要がある。次に、アダプターの一方のオリゴヌクレオチドと同一配列を有するプライマーを用いて、PCR法により増幅し、無尽蔵化することができる。一方、各BAC DNA等に特徴的な50〜70塩基のアミノ化オリゴヌクレオチドを検出用プローブとして用いることもできる。
(a)二色蛍光法
DNAアレイと標識核酸を用いるハイブリダイゼーション法[CGH(Comparative Genomic Hybridization)法]では、例えば、二色蛍光法を用いて行うことができる。
本発明において「標識」とは、検出可能な物質を標的核酸に対して結合させる行為であり、検出可能であれば、いかなるものも本発明にかかわる標的核酸に組み込むことができる。例えば、蛍光物質、無機化合物、タンパク質(ELISA(Enzyme-Linked ImmunoSorbent Assay)などで使用される酵素標識抗体など)、放射性同位元素、FRET(蛍光共鳴エネルギー移動)などを選択ことができる。
本発明において、標的核酸を検体から調製する際には、精製を行う必要がある。この場合には、後述する標識の際に、様々な副反応が生じると共に、タンパクや脂質など細胞溶解物がバックグラウンドノイズに大きな影響を与え、精製を行わなければ、核酸マイクロアレイ等を用いるハイブリダイゼーション試験の性能を著しく低下させる。
(1)染色体検査
原因不明の精神遅滞を伴う多発性奇形症候群の疾患に関してヒトゲノムの特定領域の異常が明確になれば、特に、10Mb程度の欠失を伴い10q24.31-10q25.1領域が、ヘミ接合体欠失している場合には、G分染法等の染色体検査により検出することが可能である。本発明の該当領域の染色状態の違い(10q24.31-10q25.1領域を含む染色体バンドの消失の有無)を観察することで検出することができる。
数Mb以下の微細なゲノム異常であればFISH法[蛍光in situ ハイブリダイゼーション(FISH: Fluorescence in situ hybridization):Yasui,K., Imoto,I., Fukuda,Y., Pimkhaokham,A., Yang,Z.Q., Naruto,T., Shimada,Y., Nakamura,Y., and Inazawa,J: Identification of target genes within an amplicon at 14q12-q13 in esophageal squamous cell carcinoma. Genes Chromosomes Cancer, 32, 112-118, 2001]、等により、核酸のハイブリダイゼーションによるシグナルを基に検出することが可能となる。この場合、10q24.31-10q25.1領域のFISHプローブを作成し、患者由来の染色体とハイブリダイゼーションを行い、シグナル数の減少を観察することで、ヘミ接合体欠失を検出することが可能である。
サザンブロット法を用いる場合、検体から得られるゲノムDNAを制限酵素消化し、それをゲル電気泳動後、ニトロセルロース膜上に固定し、これと、標識した10q24.32-q25.1領域のDNAとハイブリダイゼーションを行い検出することにより、検体中の当該遺伝子の存在を検出する方法である。ヘミ接合体欠失の場合、正常由来の検体から得られる検出量(バンドの濃さ)に対し、患者由来の検体から得られる検出量が少ないことと共に、異なるバンドの出現が認められることにより、判定することができる。
ノーザンブロット法は、検体から得られる全RNAを電気泳動に供し、サザンブロッティング法と同様の操作で膜上に固定し、これと、標識した10q24.32-q25.1領域のDNAとハイブリダイゼーションを行い検出することにより、検体中の当該遺伝子の存在を検出する方法である。正常由来の検体から得られる検出量(バンドの濃さ)に対し、患者由来の検体から得られる検出量が少ない場合、10q24.32-q25.1領域のヘミ接合体欠失として判定することができる場合がある。
リアルタイムRT−PCR法は、検体DNAの10q24.32-q25.1領域の転写産物に対応するプライマーを少なくとも1種類準備し逆転写反応を行い、次に、遺伝子の増幅工程を行い、当該増幅産物の生成の有無や生成量から検出する方法である。正常由来DNAの増幅産物量に対し、患者由来DNAからの増幅産物量が少ないことで、ヘミ接合体欠失として判定できる場合がある。
(A)MCG Whole Genome Array-4500
上述したように、本発明の検出方法を行うに際して、ヒト染色体の10q24.31-10q25.1の領域のヘミ接合体欠失が、精神遅滞を伴う多発性奇形症候群に関連していることを見出すために用いた検出手段は、MCG Whole Genome Array-4500である。この検出基板については公知(非特許文献1)であるが、念のために、ここにその製造過程を簡単に説明する。
(1)精神遅滞を伴う多発性奇形症候群患者2名の臨床像
本実施例において、精神遅滞を伴う多発性奇形症候群に対応するヒト染色体の領域を特定するための検体提供者2名[精神遅滞を伴う新規先天性異常症患児2名(ケース1と2)]の臨床像について、可能な範囲で開示する。
(a)Genome Disorder Arrayでの解析結果
Genome Disorder Arrayは、ヒトゲノムDNAが、先天性異常症疾患で欠失または増幅する領域のゲノムDNA(BACクローン)が定着している基板である。
再び、ケース1及び2の患者の血液からDNAを調製し、上記の要領にて製造される、MCG Whole Genome Array-4500を用いたアレイCGH法によるゲノム構造異常に関する解析を、下記の要領にて行なった。
ケース1及びケース2の患者から調製したDNA(各々0.2μg)を用いて、アジレント社のオリゴaCGHマイクロアレイ244KタイプによるCGH法での解析を行った。244Kタイプのアレイはヒトゲノムの236000以上のコード領域および非コード領域をカバーしており、ヒトゲノムの網羅的解析が可能である。プローブの平均分解能は6.4Kbである。解析の概要は次の通りでAgilent社のプロトコルに従って行った。
精神遅滞を伴う多発性奇形症候群患者染色体中に10q24.31-10q25.1領域のヘミ接合体欠失があることを、FISH法を用いて確認した。まず、本患者2名の血液よりリンパ球を調製し、メタフェーズ染色体を作製した。即ち、ヒトリンパ球を12.5μg/mlのPhytohemagglutininと共に3日間培養し、さらに、0.025μg/mlのコルセミドを添加し数時間培養した後、上清を除き、0.075M KCl低張液を加え、30分間静置した。続いて、上清を除いた後、カルノア液で固定を行い、スライドガラス上に染色体を展開しメタフェーズ染色体を作製した。
Claims (6)
- ヒト染色体の10q24.33領域のヘミ接合体欠失を検出することにより、精神遅滞を伴う多発性奇形症候群の判別を補助する、染色体欠失の検出方法。
- ヒト染色体の10q24.33領域のヘミ接合体欠失の検出が、当該10q24.33領域の一部を含む核酸と検体核酸とのハイブリダイゼーションにより、当該10q24.33領域のヘミ接合体欠失に基づいて発生するシグナルを検出することにより行われる、請求項1に記載の染色体欠失の検出方法。
- ヒト染色体の10q24.33領域の一部を含む核酸と検体核酸とのハイブリダイゼーションが、当該10q24.33の領域の一部を含む核酸が定着した基板において行われる、請求項2に記載の染色体欠失の検出方法。
- ヒト染色体の10q24.33の領域の一部を含む核酸が、オリゴヌクレオチド、cDNA、BAC DNA、PAC DNA、又は、YAC DNAである、請求項2又は3に記載の染色体欠失の検出方法。
- ヒト染色体の10q24.33の領域の一部を含む核酸が、BAC DNAであるRP11−416N2である、請求項4に記載の染色体欠失の検出方法。
- ヒト染色体の10q24.33の領域の一部又は全部を含む核酸を検出する手段が、DNAチップ法、サザンブロット法、ノーザンブロット法、リアルタイムRT−PCR法、FISH法、又は、CGH法を用いて検出する、請求項1〜5のいずれかに記載の染色体欠失の検出方法。
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2008199541A JP5331404B2 (ja) | 2008-08-01 | 2008-08-01 | 先天性異常症の染色体欠失の検出方法 |
CN200980130226.0A CN102112628B (zh) | 2008-08-01 | 2009-07-30 | 先天性异常症的染色体缺失的检测方法 |
EP09803072.9A EP2319938B1 (en) | 2008-08-01 | 2009-07-30 | Method for detecting chromosome deficiencies for congenital abnormality |
PCT/JP2009/063900 WO2010013842A1 (ja) | 2008-08-01 | 2009-07-30 | 先天性異常症の染色体欠失の検出方法 |
US13/056,935 US20110207626A1 (en) | 2008-08-01 | 2009-07-30 | Method for detecting chromosome deficiencies for congenital abnormality |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2008199541A JP5331404B2 (ja) | 2008-08-01 | 2008-08-01 | 先天性異常症の染色体欠失の検出方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2010035447A JP2010035447A (ja) | 2010-02-18 |
JP5331404B2 true JP5331404B2 (ja) | 2013-10-30 |
Family
ID=41610529
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2008199541A Active JP5331404B2 (ja) | 2008-08-01 | 2008-08-01 | 先天性異常症の染色体欠失の検出方法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US20110207626A1 (ja) |
EP (1) | EP2319938B1 (ja) |
JP (1) | JP5331404B2 (ja) |
CN (1) | CN102112628B (ja) |
WO (1) | WO2010013842A1 (ja) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8633200B2 (en) | 2010-09-08 | 2014-01-21 | Bristol-Myers Squibb Company | Inhibitors of human immunodeficiency virus replication |
US8791108B2 (en) | 2011-08-18 | 2014-07-29 | Bristol-Myers Squibb Company | Inhibitors of human immunodeficiency virus replication |
US8629276B2 (en) | 2012-02-15 | 2014-01-14 | Bristol-Myers Squibb Company | Inhibitors of human immunodeficiency virus replication |
US9034882B2 (en) | 2012-03-05 | 2015-05-19 | Bristol-Myers Squibb Company | Inhibitors of human immunodeficiency virus replication |
US9006235B2 (en) | 2012-03-06 | 2015-04-14 | Bristol-Myers Squibb Company | Inhibitors of human immunodeficiency virus replication |
US8906929B2 (en) | 2012-08-16 | 2014-12-09 | Bristol-Myers Squibb Company | Inhibitors of human immunodeficiency virus replication |
WO2015051006A2 (en) * | 2013-10-01 | 2015-04-09 | Complete Genomics, Inc. | Phasing and linking processes to identify variations in a genome |
AU2015243449A1 (en) * | 2014-04-09 | 2016-09-29 | Lineagen, Inc. | Genetic markers associated with chromosomal deletion and duplication syndromes |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2463725A1 (en) * | 2001-10-12 | 2003-11-06 | Spectral Genomics, Inc. | Compilations of nucleic acids and arrays and methods of using them |
JP2005304481A (ja) * | 2004-02-13 | 2005-11-04 | Joji Inasawa | ゲノムdnaの定着基盤と当該基盤を用いた染色体異常並びにそれに起因する疾患の検出方法 |
JP2008072939A (ja) * | 2006-09-21 | 2008-04-03 | Fujifilm Corp | 多発性骨髄腫の検出方法および抑制方法 |
-
2008
- 2008-08-01 JP JP2008199541A patent/JP5331404B2/ja active Active
-
2009
- 2009-07-30 WO PCT/JP2009/063900 patent/WO2010013842A1/ja active Application Filing
- 2009-07-30 US US13/056,935 patent/US20110207626A1/en not_active Abandoned
- 2009-07-30 CN CN200980130226.0A patent/CN102112628B/zh active Active
- 2009-07-30 EP EP09803072.9A patent/EP2319938B1/en active Active
Also Published As
Publication number | Publication date |
---|---|
JP2010035447A (ja) | 2010-02-18 |
EP2319938A1 (en) | 2011-05-11 |
EP2319938B1 (en) | 2016-06-01 |
US20110207626A1 (en) | 2011-08-25 |
EP2319938A4 (en) | 2012-07-18 |
WO2010013842A1 (ja) | 2010-02-04 |
CN102112628B (zh) | 2013-07-17 |
CN102112628A (zh) | 2011-06-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5331404B2 (ja) | 先天性異常症の染色体欠失の検出方法 | |
Cui et al. | Fluorescence in situ hybridization: cell-based genetic diagnostic and research applications | |
EP0614492B1 (en) | In situ hybridization method | |
US6828097B1 (en) | Single copy genomic hybridization probes and method of generating same | |
JP5149622B2 (ja) | 単一標識比較ハイブリダイゼーション | |
US5650277A (en) | Method of determining the presence and quantifying the number of di- and trinucleotide repeats | |
JP2005519634A (ja) | 核酸組成物及びアレイ及びそれらを用いる方法 | |
JP2007515947A (ja) | 羊水中の無細胞胎児dnaを使用する出生前診断 | |
CA2409752A1 (en) | Single copy genomic hybridization probes and method of generating same | |
AU2001264610A1 (en) | Single copy genomic hybridization probes and method of generating same | |
US20150252412A1 (en) | High-definition dna in situ hybridization (hd-fish) compositions and methods | |
US20230183780A1 (en) | Dna probes for in situ hybridization on chromosomes | |
WO2019080595A1 (zh) | 一种原位杂交探针的制备方法 | |
US8021888B2 (en) | Rapid comparative genomic hybridization using acoustic surface waves | |
JPH08507198A (ja) | 特異的対立遺伝子の検出のための単一ヌクレオチドプライマー伸張法およびそのためのキット | |
JPH07505777A (ja) | 一般的な生産の染色体異数体の検出用プローブ | |
WO2010101273A1 (ja) | 精神遅滞を伴う多発性奇形症候群の判別方法 | |
WO2008050870A1 (fr) | Gène spécifique d'un organe, procédé d'identification de celui-ci et son utilisation | |
CN111118157B (zh) | 一种长链非编码RNA lncBCBMAT及其作为乳腺癌脑转移分子标记物的应用 | |
Smajkan | Applications of Fluorescent in situ hybridization (FISH) | |
TWI564561B (zh) | Detection of KRAS oncogene for circulating cancer cells | |
CN114196788A (zh) | 利用荧光原位检测技术快速检测非洲猪瘟病毒的方法 | |
Halder et al. | Laboratory manual | |
Halling et al. | In situ hybridization: principles and applications for pulmonary medicine | |
WO2004018691A1 (fr) | Procede pour adn chromosomique micro-excise d'amplification et son utilisation |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20110721 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20110801 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20110721 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20110801 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20120925 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20121126 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20121128 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20130219 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20130328 |
|
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: 20130723 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20130729 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5331404 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 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 |