CN101967517A - 一种无需借助pcr的基因检测方法 - Google Patents
一种无需借助pcr的基因检测方法 Download PDFInfo
- Publication number
- CN101967517A CN101967517A CN201010147160.0A CN201010147160A CN101967517A CN 101967517 A CN101967517 A CN 101967517A CN 201010147160 A CN201010147160 A CN 201010147160A CN 101967517 A CN101967517 A CN 101967517A
- Authority
- CN
- China
- Prior art keywords
- dna
- probe
- molecule
- mass spectrum
- nano
- 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.)
- Granted
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 39
- 238000001514 detection method Methods 0.000 title abstract description 45
- 238000003752 polymerase chain reaction Methods 0.000 title abstract description 13
- 239000000523 sample Substances 0.000 claims abstract description 128
- 238000009396 hybridization Methods 0.000 claims abstract description 24
- 239000004005 microsphere Substances 0.000 claims abstract description 23
- 150000001793 charged compounds Chemical class 0.000 claims abstract description 4
- 239000000758 substrate Substances 0.000 claims abstract description 3
- 238000001819 mass spectrum Methods 0.000 claims description 51
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 42
- 239000000203 mixture Substances 0.000 claims description 27
- 230000002285 radioactive effect Effects 0.000 claims description 26
- 239000000126 substance Substances 0.000 claims description 16
- 239000010931 gold Substances 0.000 claims description 14
- 229910052737 gold Inorganic materials 0.000 claims description 14
- 239000011159 matrix material Substances 0.000 claims description 10
- 238000001338 self-assembly Methods 0.000 claims description 10
- 239000002245 particle Substances 0.000 claims description 9
- 239000002105 nanoparticle Substances 0.000 claims description 8
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- 210000001541 thymus gland Anatomy 0.000 claims description 7
- 230000015572 biosynthetic process Effects 0.000 claims description 6
- 239000011782 vitamin Substances 0.000 claims description 5
- 229940088594 vitamin Drugs 0.000 claims description 5
- 229930003231 vitamin Natural products 0.000 claims description 5
- 235000013343 vitamin Nutrition 0.000 claims description 5
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 5
- 239000004793 Polystyrene Substances 0.000 claims description 4
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 108010090804 Streptavidin Proteins 0.000 claims description 3
- 229920001222 biopolymer Polymers 0.000 claims description 3
- 229920000642 polymer Polymers 0.000 claims description 3
- ZTFXOSFQSNZIFS-UHFFFAOYSA-N 3-(3,5-diethoxy-4-hydroxyphenyl)prop-2-enoic acid Chemical compound CCOc1cc(C=CC(O)=O)cc(OCC)c1O ZTFXOSFQSNZIFS-UHFFFAOYSA-N 0.000 claims description 2
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 claims description 2
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 claims description 2
- 238000000119 electrospray ionisation mass spectrum Methods 0.000 claims description 2
- 150000002894 organic compounds Chemical class 0.000 claims description 2
- 150000003230 pyrimidines Chemical class 0.000 claims description 2
- PCMORTLOPMLEFB-ONEGZZNKSA-N sinapic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC(OC)=C1O PCMORTLOPMLEFB-ONEGZZNKSA-N 0.000 claims description 2
- PCMORTLOPMLEFB-UHFFFAOYSA-N sinapinic acid Natural products COC1=CC(C=CC(O)=O)=CC(OC)=C1O PCMORTLOPMLEFB-UHFFFAOYSA-N 0.000 claims description 2
- 150000003568 thioethers Chemical class 0.000 claims description 2
- AFVLVVWMAFSXCK-UHFFFAOYSA-N α-cyano-4-hydroxycinnamic acid Chemical group OC(=O)C(C#N)=CC1=CC=C(O)C=C1 AFVLVVWMAFSXCK-UHFFFAOYSA-N 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 49
- 230000035945 sensitivity Effects 0.000 abstract description 27
- 238000005406 washing Methods 0.000 abstract description 26
- 239000012472 biological sample Substances 0.000 abstract description 14
- 201000010099 disease Diseases 0.000 abstract description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 12
- 238000003745 diagnosis Methods 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 5
- 238000013399 early diagnosis Methods 0.000 abstract description 5
- 238000001269 time-of-flight mass spectrometry Methods 0.000 abstract description 2
- 238000003795 desorption Methods 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 101
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 88
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 44
- 239000000872 buffer Substances 0.000 description 42
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 38
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical class CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 26
- 235000002639 sodium chloride Nutrition 0.000 description 24
- 239000011734 sodium Substances 0.000 description 21
- 230000008569 process Effects 0.000 description 19
- 239000011780 sodium chloride Substances 0.000 description 18
- 238000006243 chemical reaction Methods 0.000 description 17
- 239000000243 solution Substances 0.000 description 16
- 238000012360 testing method Methods 0.000 description 15
- 230000008030 elimination Effects 0.000 description 14
- 238000003379 elimination reaction Methods 0.000 description 14
- 239000012535 impurity Substances 0.000 description 14
- 238000002474 experimental method Methods 0.000 description 13
- 239000000047 product Substances 0.000 description 11
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 10
- 230000001133 acceleration Effects 0.000 description 10
- 150000002500 ions Chemical class 0.000 description 10
- 238000004949 mass spectrometry Methods 0.000 description 10
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 10
- 239000011259 mixed solution Substances 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 10
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 8
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 239000003643 water by type Substances 0.000 description 7
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 6
- 229910052708 sodium Inorganic materials 0.000 description 6
- MYLBTCQBKAKUTJ-UHFFFAOYSA-N 7-methyl-6,8-bis(methylsulfanyl)pyrrolo[1,2-a]pyrazine Chemical compound C1=CN=CC2=C(SC)C(C)=C(SC)N21 MYLBTCQBKAKUTJ-UHFFFAOYSA-N 0.000 description 5
- 108091034117 Oligonucleotide Proteins 0.000 description 5
- 238000012408 PCR amplification Methods 0.000 description 5
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 5
- 239000011324 bead Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 229910052717 sulfur Inorganic materials 0.000 description 5
- 239000011593 sulfur Substances 0.000 description 5
- 229910021642 ultra pure water Inorganic materials 0.000 description 5
- 239000012498 ultrapure water Substances 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 241000725303 Human immunodeficiency virus Species 0.000 description 4
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 4
- 239000003550 marker Substances 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 229910052710 silicon Inorganic materials 0.000 description 4
- 239000010703 silicon Substances 0.000 description 4
- 229960002668 sodium chloride Drugs 0.000 description 4
- IBUIVNCCBFLEJL-UHFFFAOYSA-M sodium;phosphoric acid;chloride Chemical class [Na+].[Cl-].OP(O)(O)=O IBUIVNCCBFLEJL-UHFFFAOYSA-M 0.000 description 4
- KZNICNPSHKQLFF-UHFFFAOYSA-N succinimide Chemical compound O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 description 4
- PMJWDPGOWBRILU-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 4-[4-(2,5-dioxopyrrol-1-yl)phenyl]butanoate Chemical compound O=C1CCC(=O)N1OC(=O)CCCC(C=C1)=CC=C1N1C(=O)C=CC1=O PMJWDPGOWBRILU-UHFFFAOYSA-N 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 3
- 102100033072 DNA replication ATP-dependent helicase DNA2 Human genes 0.000 description 3
- 101000927313 Homo sapiens DNA replication ATP-dependent helicase DNA2 Proteins 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 238000002161 passivation Methods 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 2
- ZAKOWWREFLAJOT-CEFNRUSXSA-N D-alpha-tocopherylacetate Chemical compound CC(=O)OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-CEFNRUSXSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 2
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 2
- 229910014103 Na-S Inorganic materials 0.000 description 2
- 229910014147 Na—S Inorganic materials 0.000 description 2
- 208000019802 Sexually transmitted disease Diseases 0.000 description 2
- 108020004682 Single-Stranded DNA Proteins 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000006062 fragmentation reaction Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000006249 magnetic particle Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000002086 nanomaterial Substances 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000004153 renaturation Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 229960002317 succinimide Drugs 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- SNBCLPGEMZEWLU-QXFUBDJGSA-N 2-chloro-n-[[(2r,3s,5r)-3-hydroxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methyl]acetamide Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CNC(=O)CCl)[C@@H](O)C1 SNBCLPGEMZEWLU-QXFUBDJGSA-N 0.000 description 1
- 102100021391 Cationic amino acid transporter 3 Human genes 0.000 description 1
- 206010008909 Chronic Hepatitis Diseases 0.000 description 1
- 108020004638 Circular DNA Proteins 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 108010005054 Deoxyribonuclease BamHI Proteins 0.000 description 1
- 102100031780 Endonuclease Human genes 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 101100175482 Glycine max CG-3 gene Proteins 0.000 description 1
- 241000711549 Hepacivirus C Species 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 108091006230 SLC7A3 Proteins 0.000 description 1
- 101150009841 TP gene Proteins 0.000 description 1
- 241000218636 Thuja Species 0.000 description 1
- 241000589884 Treponema pallidum Species 0.000 description 1
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000001311 chemical methods and process Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 229910021505 gold(III) hydroxide Inorganic materials 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000001524 infective effect Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000003771 laboratory diagnosis Methods 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000816 matrix-assisted laser desorption--ionisation Methods 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- MVMXJBMAGBRAHD-UHFFFAOYSA-N picoperine Chemical compound C=1C=CC=NC=1CN(C=1C=CC=CC=1)CCN1CCCCC1 MVMXJBMAGBRAHD-UHFFFAOYSA-N 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 208000006379 syphilis Diseases 0.000 description 1
- 239000011882 ultra-fine particle Substances 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/0027—Methods for using particle spectrometers
-
- 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
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
Abstract
Description
序列名 | 名称 | 序列 |
SEQ ID NO.5 | HIV-AuNP | 5’-GCT GTC CCT GTA ATA AAC CCG AAA ATTTTT TTT TT-(CH2)3-SH-3’ |
SEQ ID NO.6 | HBV-AuNP | 5’-CTC TGT GGT ATT GTG AGG ATT CTT GTCATT TTT TTT TT-(CH2)3-SH-3’ |
SEQ ID NO.7 | HCV-AuNP | 5’-CGC TTT CTG CGT GAA GAC AGT AGT TTTTTT TTT TT-(CH2)3-SH-3’ |
SEQ ID NO.8 | TP-AuNP | 5’-GTG TAC TAG CCC TCC CTT CTA CCT GATTTT TTT TTT-(CH2)3-SH-3’ |
SEQ ID NO.9 | HIV-MMP | 5’-SH-(CH2)6-TTT TTT TTT TTT GTA TGT CTGTTG CTA TTA TGT CTA TTA TTC TTT CCC CTG C-3’ |
SEQ ID NO.10 | HBV-MMP | 5’-SH-(CH2)6-TTT TTT TTT TCA AAC GGG CAACAT ACC TTG GTA GTC CAG AA-3’ |
SEQ ID NO.11 | HCV-MMP | 5’-SH-(CH2)6-TTT TTT TTT TCG CAA GCA CCCTAT CAG GCA GTA CCA CAA-3’ |
SEQ ID NO.12 | TP-MMP | 5’-SH-(CH2)6-TTT TTT TTT TTT TGT AAT GTATCG TTT GTT GCT TCT GTA TCT ATT TCT TGC-3’ |
SEQ ID NO.13 | 钝化DNA | 5’-SH-(CH2)6-TTT TTT TTT T-3’ |
SEQ ID NO.14 | HIV-target | 5’-TTT TCG GGT TTA TTA CAG GGA CAG C-GCAGGG GAA AGA ATA ATA GAC ATA ATA GCA ACAGAC ATA CAA-3’ |
SEQ ID NO.15 | HBV-target | 5’-TGA CAA GAA TCC TCA CAA TAC CAC AGA G-TTC TGG ACT ACC AAG GTA TGT TGC CCG TTTG-3’ |
SEQ ID NO.16 | HCV-target | 5’-AAC TAC TGT CTT CAC GCA GAA AGC G-TTGTGG TAC TGC CTG ATA GGG TGC TTG CG-3’ |
SEQ ID NO.17 | TP-target | 5’-TCA GGT AGA AGG GAG GGC TAG TAC AC-GCA AGA AAT AGA TAC AGA AGC AAC AAA CGATAC ATT ACA AA-3’ |
SEQ ID NO.18 | HGV-DNA | 5’-CAG GGT TGG TAG GTC GTA AAT CC-CCT ATTGGT CAA GAG AGA CAT-3’ |
序列名 | 名称 | 序列 |
SEQ ID NO.22 | DNAII-MMP | 5’-TAA CAA TAA CCA AAA AAA AAA A-(CH2)3SH-3’ |
SEQ ID NO.23 | DNA 1-target | 5’GGA TTA TTG TTA AAT ATT GAT AAG GAT 3’ |
SEQ ID NO.24 | DNA2-target | 5’GGA TAA TTG TTAAAT ATT GAT AAG GAT 3’ |
SEQ ID NO.25 | DNA3-target | 5’GGA TTA TTG TTA AAT ATT GAT AGG GAT 3’ |
SEQ ID NO.26 | DNAI-AuNP | 5’-HS(CH2)6-A AAA AAA AAA ATC CTT ATC AATATT-3’ |
序列名 | 名称 | 序列 |
SEQ ID NO.22 | DNAII-MMP | 5’-TAA CAA TAA CCA AAA AAA AAA A-(CH2)3SH-3’ |
SEQ ID NO.23 | DNA1-target | 5’GGA TTA TTG TTA AAT ATT GAT AAG GAT 3’ |
SEQ ID NO.26 | DNAI-AuNP | 5’-HS(CH2)6-A AAA AAA AAA ATC CTT ATC AATATT-3’ |
SEQ ID NO.27 | DNAII’-MMP | 5’-CAT ACT AAC ATA AAA AAA AAA A-(CH2)3SH-3’ |
SEQ ID NO.28 | DNA4-target | 5’TAT GTT AGT ATG ATA TAG GAA TAG TTA 3’ |
SEQ ID NO.29 | DNAI’-AuNP | 5’-HS(CH2)6-A AAA AAA AAA TAA CTA TTC CTATAT-3’ |
编号 | 编码分子与识别分子的浓度比例 | 编码分子用量(μl) | 识别分子用量 |
1a | 150∶1 | 45 | 3.0nmol |
2a | 200∶1 | 60 | 3.0nmol |
3a | 250∶1 | 75 | 3.0nmol |
4a | 350∶1 | 105 | 3.0nmol |
5b | 500∶1 | 30 | 3.0nmol |
6b | 800∶1 | 50 | 3.0nmol |
7b | 1000∶1 | 60 | 3.0nmol |
8b | 1300∶1 | 80 | 3.0nmol |
9b | 1700∶1 | 100 | 3.0nmol |
10c | 2000∶1 | 60 | 3.0nmol |
11c | 2300∶1 | 70 | 3.0nmol |
12c | 2700∶1 | 80 | 3.0nmol |
13c | 3000∶1 | 90 | 3.0nmol |
14c | 3300∶1 | 100 | 3.0nmol |
15c | 3700∶1 | 110 | 3.0nmol |
Claims (10)
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201010147160.0A CN101967517B (zh) | 2010-03-19 | 2010-03-19 | 一种无需借助pcr的基因检测方法 |
US13/635,879 US8704165B2 (en) | 2010-03-19 | 2011-02-17 | Gene detecting methods without using PCR |
PCT/CN2011/071044 WO2011113313A1 (zh) | 2010-03-19 | 2011-02-17 | 无需使用pcr的基因检测方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201010147160.0A CN101967517B (zh) | 2010-03-19 | 2010-03-19 | 一种无需借助pcr的基因检测方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN101967517A true CN101967517A (zh) | 2011-02-09 |
CN101967517B CN101967517B (zh) | 2012-11-07 |
Family
ID=43546739
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201010147160.0A Active CN101967517B (zh) | 2010-03-19 | 2010-03-19 | 一种无需借助pcr的基因检测方法 |
Country Status (3)
Country | Link |
---|---|
US (1) | US8704165B2 (zh) |
CN (1) | CN101967517B (zh) |
WO (1) | WO2011113313A1 (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011113313A1 (zh) * | 2010-03-19 | 2011-09-22 | 东英(江苏)药业有限公司 | 无需使用pcr的基因检测方法 |
CN109342732A (zh) * | 2018-10-31 | 2019-02-15 | 中国农业科学院油料作物研究所 | 一种结合诱导dna组装荧光检测器件及利用其检测小分子含量的方法 |
CN113355398A (zh) * | 2021-04-23 | 2021-09-07 | 厦门大学 | 一种基于BCA技术以Au NP为基质的MALDI MS检测DNA/蛋白质的方法 |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10876152B2 (en) | 2012-09-04 | 2020-12-29 | Guardant Health, Inc. | Systems and methods to detect rare mutations and copy number variation |
US20160040229A1 (en) | 2013-08-16 | 2016-02-11 | Guardant Health, Inc. | Systems and methods to detect rare mutations and copy number variation |
EP2893040B1 (en) | 2012-09-04 | 2019-01-02 | Guardant Health, Inc. | Methods to detect rare mutations and copy number variation |
US11913065B2 (en) | 2012-09-04 | 2024-02-27 | Guardent Health, Inc. | Systems and methods to detect rare mutations and copy number variation |
CN111534580A (zh) | 2013-12-28 | 2020-08-14 | 夸登特健康公司 | 用于检测遗传变异的方法和系统 |
EP3390668A4 (en) | 2015-12-17 | 2020-04-01 | Guardant Health, Inc. | METHODS OF DETERMINING THE NUMBER OF TUMOR GENE COPIES BY ACELLULAR DNA ANALYSIS |
CN111218496B (zh) * | 2020-01-16 | 2023-06-27 | 中南大学 | 一种基于dna步行者偶联的磁性纳米复合物的制备方法及其产品与应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1463291A (zh) * | 2001-04-19 | 2003-12-24 | 赛弗根生物系统股份有限公司 | 用质谱法和亲和标记物鉴定生物分子的特性 |
WO2007002567A2 (en) * | 2005-06-23 | 2007-01-04 | Nanosphere, Inc. | Selective isolation and concentration of nucleic acids from complex samples |
WO2007024676A2 (en) * | 2005-08-19 | 2007-03-01 | Nanosphere, Inc. | Methods for preparing hybrid substrates comprising dna and antibodies and uses thereof |
CN101281164A (zh) * | 2008-02-27 | 2008-10-08 | 南京大学 | 制备双分子修饰的纳米探针的方法及应用 |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050250094A1 (en) * | 2003-05-30 | 2005-11-10 | Nanosphere, Inc. | Method for detecting analytes based on evanescent illumination and scatter-based detection of nanoparticle probe complexes |
JP4557973B2 (ja) * | 2004-03-31 | 2010-10-06 | 塩野義製薬株式会社 | 金属微粒子を用いた質量分析方法 |
US20110014632A1 (en) * | 2008-12-08 | 2011-01-20 | Nanosphere, Inc. | Assays for clinical assessments of rheumatoid arthritis |
EP2389681B1 (en) * | 2009-01-23 | 2019-08-21 | Ionwerks, Inc. | Post-ionization of neutrals for ion mobility otofms identification of molecules and elements desorbed from surfaces |
US20110171749A1 (en) * | 2009-03-02 | 2011-07-14 | Board Of Trustees Of Michigan State University | Nanoparticle tracer-based electrochemical dna sensor for detection of pathogens-amplification by a universal nano-tracer (aunt) |
CN101967517B (zh) * | 2010-03-19 | 2012-11-07 | 黄乐群 | 一种无需借助pcr的基因检测方法 |
-
2010
- 2010-03-19 CN CN201010147160.0A patent/CN101967517B/zh active Active
-
2011
- 2011-02-17 US US13/635,879 patent/US8704165B2/en active Active
- 2011-02-17 WO PCT/CN2011/071044 patent/WO2011113313A1/zh active Application Filing
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1463291A (zh) * | 2001-04-19 | 2003-12-24 | 赛弗根生物系统股份有限公司 | 用质谱法和亲和标记物鉴定生物分子的特性 |
WO2007002567A2 (en) * | 2005-06-23 | 2007-01-04 | Nanosphere, Inc. | Selective isolation and concentration of nucleic acids from complex samples |
WO2007024676A2 (en) * | 2005-08-19 | 2007-03-01 | Nanosphere, Inc. | Methods for preparing hybrid substrates comprising dna and antibodies and uses thereof |
CN101281164A (zh) * | 2008-02-27 | 2008-10-08 | 南京大学 | 制备双分子修饰的纳米探针的方法及应用 |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011113313A1 (zh) * | 2010-03-19 | 2011-09-22 | 东英(江苏)药业有限公司 | 无需使用pcr的基因检测方法 |
CN109342732A (zh) * | 2018-10-31 | 2019-02-15 | 中国农业科学院油料作物研究所 | 一种结合诱导dna组装荧光检测器件及利用其检测小分子含量的方法 |
CN109342732B (zh) * | 2018-10-31 | 2022-04-15 | 中国农业科学院油料作物研究所 | 一种结合诱导dna组装荧光检测器件及利用其检测小分子含量的方法 |
CN113355398A (zh) * | 2021-04-23 | 2021-09-07 | 厦门大学 | 一种基于BCA技术以Au NP为基质的MALDI MS检测DNA/蛋白质的方法 |
Also Published As
Publication number | Publication date |
---|---|
US20130140451A1 (en) | 2013-06-06 |
WO2011113313A1 (zh) | 2011-09-22 |
CN101967517B (zh) | 2012-11-07 |
US8704165B2 (en) | 2014-04-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101967517B (zh) | 一种无需借助pcr的基因检测方法 | |
DE69735445T2 (de) | Abspaltbare, nicht-flüchtige moleküle zur massenmarkierung | |
Han et al. | Absolute and relative quantification of multiplex DNA assays based on an elemental labeling strategy | |
US9051608B2 (en) | Detection and quantification of biomolecules using mass spectrometry | |
US8133701B2 (en) | Detection and quantification of biomolecules using mass spectrometry | |
CN102676508A (zh) | 基于纳米金和适配体的小分子探针及其制备方法 | |
Chen et al. | Elemental mass spectrometry and fluorescence dual-mode strategy for ultrasensitive label-free detection of HBV DNA | |
JP2019107016A (ja) | 少数バリアントの検出および定量のための方法マルチプレックス法 | |
CN101281164B (zh) | 制备双分子修饰的纳米探针的方法及应用 | |
CN107354207B (zh) | 基于双链探针的液相杂交捕获试剂盒、洗涤试剂盒及其应用 | |
CN112292459A (zh) | 用于核酸检测和定量的产品和方法 | |
Zhao et al. | Novel ionic liquid matrices for qualitative and quantitative detection of carbohydrates by matrix assisted laser desorption/ionization mass spectrometry | |
US7642086B2 (en) | Labeled probe bound object, method for producing the same and method for using the same | |
CN108034770A (zh) | 质谱法检测甲型流感病毒h7n9多重pcr产物的方法及其产品 | |
JP6694635B2 (ja) | マイクロrnaにおけるメチル化修飾部位を計測する方法 | |
EP1106702A1 (en) | High-throughput screening of compounds using electrospray ionization mass spectrometry (ESI-MS) | |
Yang et al. | PCR-free MDR1 polymorphism identification by gold nanoparticle probes | |
Song et al. | Quantum dot-based isothermal chain elongation for fluorescence detection of specific DNA sequences via template-dependent surface-hybridization | |
Hu et al. | Label-free detection of biomolecules using inductively coupled plasma mass spectrometry (ICP-MS) | |
CN108359744A (zh) | 检测h3n2片段多重pcr产物的质谱方法及其产品 | |
CN108034769A (zh) | 质谱检测甲型流感病毒h3n2多重pcr产物的方法及其产品 | |
CN117286140A (zh) | 一种探针组合及其构建靶向细胞原位成像的方法和应用 | |
CN117551814A (zh) | 一种检测禽流感病毒的生物传感器及检测方法和应用 | |
Ibáñez | Development of a polymeric planar microwell device (pMALDI chip) for enhancing protein analysis in combination with MALDI-TOF, MS instrumentation | |
Ibañez Gabilondo | Development of a polymeric planar microwell device (pMALDI chip) for enhancing protein analysis in combination with MALDI-TOF/MS instrumentation |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
ASS | Succession or assignment of patent right |
Owner name: JIANGSU JIAYI PHARMACEUTICAL CO., LTD. Free format text: FORMER OWNER: HUANG LEQUN Effective date: 20150206 |
|
C41 | Transfer of patent application or patent right or utility model | ||
COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 210016 NANJING, JIANGSU PROVINCE TO: 226000 NANTONG, JIANGSU PROVINCE |
|
TR01 | Transfer of patent right |
Effective date of registration: 20150206 Address after: 226000 Jiangsu city of Nantong Province Economic and Technological Development Zone of Nantong Road No. 29. Patentee after: JIANGSU YIJIA PHARMACEUTICAL CO., LTD. Address before: 210016, A, Building 29, building 2, Whampoa science and technology building, Huangpu Road, Nanjing, Jiangsu Patentee before: Huang Lequn |
|
EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20110209 Assignee: NANJING MEIJIA NINGYI MEDICINE RESEARCH DEVELOPMENT CO., LTD. Assignor: JIANGSU YIJIA PHARMACEUTICAL CO., LTD. Contract record no.: 2015320000580 Denomination of invention: Polymerase chain reaction (PCR)-free gene detection method Granted publication date: 20121107 License type: Exclusive License Record date: 20150902 |
|
LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
EC01 | Cancellation of recordation of patent licensing contract |
Assignee: NANJING MEIJIA NINGYI MEDICINE RESEARCH DEVELOPMENT CO., LTD. Assignor: JIANGSU YIJIA PHARMACEUTICAL CO., LTD. Contract record no.: 2015320000580 Date of cancellation: 20170612 |
|
EC01 | Cancellation of recordation of patent licensing contract |