CN213951203U - Rapid nucleic acid detection system and device - Google Patents
Rapid nucleic acid detection system and device Download PDFInfo
- Publication number
- CN213951203U CN213951203U CN202021381274.7U CN202021381274U CN213951203U CN 213951203 U CN213951203 U CN 213951203U CN 202021381274 U CN202021381274 U CN 202021381274U CN 213951203 U CN213951203 U CN 213951203U
- Authority
- CN
- China
- Prior art keywords
- nucleic acid
- cavity
- sealing structure
- chamber
- sample
- 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.)
- Expired - Fee Related
Links
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 128
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 127
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 127
- 238000001514 detection method Methods 0.000 title claims abstract description 61
- 230000003321 amplification Effects 0.000 claims abstract description 42
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 42
- 238000006243 chemical reaction Methods 0.000 claims abstract description 31
- 239000006166 lysate Substances 0.000 claims abstract description 6
- 238000007789 sealing Methods 0.000 claims description 101
- 239000003153 chemical reaction reagent Substances 0.000 claims description 62
- 239000000463 material Substances 0.000 claims description 19
- 230000007246 mechanism Effects 0.000 claims description 14
- 239000012188 paraffin wax Substances 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 230000009471 action Effects 0.000 claims description 8
- 239000010408 film Substances 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 8
- 238000004807 desolvation Methods 0.000 claims description 7
- 230000006378 damage Effects 0.000 claims description 6
- 230000000903 blocking effect Effects 0.000 claims description 4
- 238000002844 melting Methods 0.000 claims description 4
- 230000008018 melting Effects 0.000 claims description 4
- 238000004026 adhesive bonding Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 230000000149 penetrating effect Effects 0.000 claims description 3
- 239000011148 porous material Substances 0.000 claims description 3
- 230000007423 decrease Effects 0.000 claims description 2
- 238000002955 isolation Methods 0.000 claims description 2
- 239000010409 thin film Substances 0.000 claims description 2
- 239000011888 foil Substances 0.000 claims 1
- 241000711573 Coronaviridae Species 0.000 abstract description 12
- 238000000605 extraction Methods 0.000 abstract description 6
- 230000004308 accommodation Effects 0.000 abstract 2
- 230000000392 somatic effect Effects 0.000 abstract 2
- 239000000523 sample Substances 0.000 description 79
- 238000004891 communication Methods 0.000 description 19
- 238000000034 method Methods 0.000 description 17
- 239000000203 mixture Substances 0.000 description 16
- 239000000243 solution Substances 0.000 description 13
- 102000004190 Enzymes Human genes 0.000 description 10
- 108090000790 Enzymes Proteins 0.000 description 10
- 238000010586 diagram Methods 0.000 description 10
- 238000003752 polymerase chain reaction Methods 0.000 description 9
- 230000008569 process Effects 0.000 description 9
- 230000008859 change Effects 0.000 description 8
- 230000009089 cytolysis Effects 0.000 description 8
- 238000003745 diagnosis Methods 0.000 description 7
- 238000006073 displacement reaction Methods 0.000 description 7
- 239000002904 solvent Substances 0.000 description 7
- 241001678559 COVID-19 virus Species 0.000 description 6
- 238000007397 LAMP assay Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 238000010839 reverse transcription Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 108091093088 Amplicon Proteins 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 108020004463 18S ribosomal RNA Proteins 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 210000003296 saliva Anatomy 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 3
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 3
- 239000000980 acid dye Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000007664 blowing Methods 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- -1 enzyme) Chemical class 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 241000711467 Human coronavirus 229E Species 0.000 description 2
- 241000482741 Human coronavirus NL63 Species 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 241000315672 SARS coronavirus Species 0.000 description 2
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 2
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 159000000003 magnesium salts Chemical class 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 238000005192 partition Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 208000011580 syndromic disease Diseases 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000005758 transcription activity Effects 0.000 description 2
- KJCVRFUGPWSIIH-UHFFFAOYSA-N 1-naphthol Chemical compound C1=CC=C2C(O)=CC=CC2=C1 KJCVRFUGPWSIIH-UHFFFAOYSA-N 0.000 description 1
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- RTZZCYNQPHTPPL-UHFFFAOYSA-N 3-nitrophenol Chemical compound OC1=CC=CC([N+]([O-])=O)=C1 RTZZCYNQPHTPPL-UHFFFAOYSA-N 0.000 description 1
- OLQIKGSZDTXODA-UHFFFAOYSA-N 4-[3-(4-hydroxy-2-methylphenyl)-1,1-dioxo-2,1$l^{6}-benzoxathiol-3-yl]-3-methylphenol Chemical compound CC1=CC(O)=CC=C1C1(C=2C(=CC(O)=CC=2)C)C2=CC=CC=C2S(=O)(=O)O1 OLQIKGSZDTXODA-UHFFFAOYSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- CEZCCHQBSQPRMU-LLIZZRELSA-L Allura red AC Chemical compound [Na+].[Na+].COC1=CC(S([O-])(=O)=O)=C(C)C=C1\N=N\C1=C(O)C=CC2=CC(S([O-])(=O)=O)=CC=C12 CEZCCHQBSQPRMU-LLIZZRELSA-L 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-N Betaine Natural products C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000411951 Centrosema virginianum Species 0.000 description 1
- 235000008358 Clitoria ternatea Nutrition 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 108020004635 Complementary DNA Proteins 0.000 description 1
- 102000016911 Deoxyribonucleases Human genes 0.000 description 1
- 108010053770 Deoxyribonucleases Proteins 0.000 description 1
- AHCYMLUZIRLXAA-SHYZEUOFSA-N Deoxyuridine 5'-triphosphate Chemical compound O1[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C[C@@H]1N1C(=O)NC(=O)C=C1 AHCYMLUZIRLXAA-SHYZEUOFSA-N 0.000 description 1
- 229920002943 EPDM rubber Polymers 0.000 description 1
- 239000000940 FEMA 2235 Substances 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 244000309467 Human Coronavirus Species 0.000 description 1
- 241001109669 Human coronavirus HKU1 Species 0.000 description 1
- 241001428935 Human coronavirus OC43 Species 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 229920000265 Polyparaphenylene Polymers 0.000 description 1
- 229920000491 Polyphenylsulfone Polymers 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H 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
- 241000907663 Siproeta stelenes Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- OIQPTROHQCGFEF-QIKYXUGXSA-L Sunset Yellow FCF Chemical compound [Na+].[Na+].OC1=CC=C2C=C(S([O-])(=O)=O)C=CC2=C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 OIQPTROHQCGFEF-QIKYXUGXSA-L 0.000 description 1
- 108010020713 Tth polymerase Proteins 0.000 description 1
- 235000009499 Vanilla fragrans Nutrition 0.000 description 1
- 235000012036 Vanilla tahitensis Nutrition 0.000 description 1
- 244000263375 Vanilla tahitensis Species 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 235000012741 allura red AC Nutrition 0.000 description 1
- 239000004191 allura red AC Substances 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- IRERQBUNZFJFGC-UHFFFAOYSA-L azure blue Chemical compound [Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Al+3].[Al+3].[Al+3].[Al+3].[Al+3].[Al+3].[S-]S[S-].[O-][Si]([O-])([O-])[O-].[O-][Si]([O-])([O-])[O-].[O-][Si]([O-])([O-])[O-].[O-][Si]([O-])([O-])[O-].[O-][Si]([O-])([O-])[O-].[O-][Si]([O-])([O-])[O-] IRERQBUNZFJFGC-UHFFFAOYSA-L 0.000 description 1
- 229960003237 betaine Drugs 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229930002875 chlorophyll Natural products 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- 229940106705 chlorophyll Drugs 0.000 description 1
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- OBRMNDMBJQTZHV-UHFFFAOYSA-N cresol red Chemical group C1=C(O)C(C)=CC(C2(C3=CC=CC=C3S(=O)(=O)O2)C=2C=C(C)C(O)=CC=2)=C1 OBRMNDMBJQTZHV-UHFFFAOYSA-N 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 1
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 1
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 1
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000004205 dimethyl polysiloxane Substances 0.000 description 1
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- IINNWAYUJNWZRM-UHFFFAOYSA-L erythrosin B Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=C(I)C(=O)C(I)=C2OC2=C(I)C([O-])=C(I)C=C21 IINNWAYUJNWZRM-UHFFFAOYSA-L 0.000 description 1
- 235000012732 erythrosine Nutrition 0.000 description 1
- 239000004174 erythrosine Substances 0.000 description 1
- 229940011411 erythrosine Drugs 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 238000007731 hot pressing Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- KHLVKKOJDHCJMG-QDBORUFSSA-L indigo carmine Chemical compound [Na+].[Na+].N/1C2=CC=C(S([O-])(=O)=O)C=C2C(=O)C\1=C1/NC2=CC=C(S(=O)(=O)[O-])C=C2C1=O KHLVKKOJDHCJMG-QDBORUFSSA-L 0.000 description 1
- 229960003988 indigo carmine Drugs 0.000 description 1
- 235000012738 indigotine Nutrition 0.000 description 1
- 239000004179 indigotine Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011901 isothermal amplification Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000155 melt Substances 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- PGSADBUBUOPOJS-UHFFFAOYSA-N neutral red Chemical compound Cl.C1=C(C)C(N)=CC2=NC3=CC(N(C)C)=CC=C3N=C21 PGSADBUBUOPOJS-UHFFFAOYSA-N 0.000 description 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 1
- 229960002378 oftasceine Drugs 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- IEQIEDJGQAUEQZ-UHFFFAOYSA-N phthalocyanine Chemical compound N1C(N=C2C3=CC=CC=C3C(N=C3C4=CC=CC=C4C(=N4)N3)=N2)=C(C=CC=C2)C2=C1N=C1C2=CC=CC=C2C4=N1 IEQIEDJGQAUEQZ-UHFFFAOYSA-N 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 229920001200 poly(ethylene-vinyl acetate) Polymers 0.000 description 1
- 229920000139 polyethylene terephthalate Polymers 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000037452 priming Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 235000012751 sunset yellow FCF Nutrition 0.000 description 1
- 239000004173 sunset yellow FCF Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 235000012756 tartrazine Nutrition 0.000 description 1
- 239000004149 tartrazine Substances 0.000 description 1
- 229960000943 tartrazine Drugs 0.000 description 1
- UJMBCXLDXJUMFB-GLCFPVLVSA-K tartrazine Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)C1=NN(C=2C=CC(=CC=2)S([O-])(=O)=O)C(=O)C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 UJMBCXLDXJUMFB-GLCFPVLVSA-K 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- AUIINJJXRXMPGT-UHFFFAOYSA-K trisodium 3-hydroxy-4-[(2-hydroxy-4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2,7-disulfonate Chemical compound [Na+].[Na+].[Na+].Oc1cc(c2ccccc2c1N=Nc1c(O)c(cc2cc(ccc12)S([O-])(=O)=O)S([O-])(=O)=O)S([O-])(=O)=O AUIINJJXRXMPGT-UHFFFAOYSA-K 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The utility model discloses a novel coronavirus's quick nucleic acid detecting system and nucleic acid detecting device, be in including the somatic part and the setting that have accommodation space can keep apart the seal structure that can communicate on this somatic part, seal structure will accommodation space separates into the first cavity that is used for collecting the sample, is used for holding the second cavity of lysate and is used for carrying out the third cavity of nucleic acid amplification reaction. The system integrates sample collection, nucleic acid extraction, reaction and observation, does not need special instruments, is convenient to sample, has simple operation of a detection device of the system, can detect the novel coronavirus quickly, accurately and conveniently, and has good application prospect.
Description
Technical Field
The utility model relates to an external diagnosis field, concretely relates to quick nucleic acid detecting system and device.
Background
Nucleic acid detection, also known as molecular detection, is a diagnostic by detecting the genetic material nucleic acid of viruses, and is a gold standard for the diagnosis of various viral infectious diseases; for example, the method can be used for detecting Coronavirus (CoV), which is a pathogenic microorganism seriously harming human and livestock health and is commonly found in nature, natural hosts comprise human, cattle, pig, dog, bird, cat, bat, rat and the like, and the genetic change is easy to occur in the evolution process due to the wide host characteristics and the self genome structure, so that the genetic diversity is presented, and new coronaviruses and subtypes continuously appear. The number of coronavirus known to infect humans is 7, namely human coronavirus 229E (HCoV-229E) and OC43(HCoV-OC43) discovered in the 60 th generation of the 20 th century, SARS coronavirus newly appearing in 2003 (sever acuterpityriasis syndrome virus, SARS-CoV), human coronavirus NL63(HCoV-NL63) discovered in 2004, human coronavirus hong kong I (HCoV-HKU1) newly discovered in 2005, middle east respiratory syndrome coronavirus (MERS-CoV) appearing in the middle east area in 2012, and novel coronavirus (sever acute respiratory syndrome coronavirus 2, SARS-CoV-2) in 2020.
The SARS-CoV-2 nucleic acid detection can provide direct evidence for diagnosis, and in the current latest version of diagnosis and treatment scheme, the detection of SARS-CoV-2 nucleic acid is still the only basis for diagnosis. Compared with immunoassay, the nucleic acid detection has high sensitivity, strong specificity and short window period, and is generally 3-5 days (theoretically, nucleic acid can be detected only by infecting mucosa). However, since nucleic acid detection usually requires four steps of sampling, inactivation, nucleic acid extraction, and detection on a computer, the detection time is long, especially when the nucleic acid detection technology used is the most common Polymerase Chain Reaction (PCR) in the industry at present, it takes more than 6 hours for a batch of samples to be tested and reported finally. If the PCR technology is used for detecting the nucleic acid of the new coronavirus, the detection result cannot be timely and effectively obtained, so that the response speed of preventing and controlling the epidemic situation can be influenced to a certain extent. Although the nucleic acid kit has an unsatisfactory place at present, the nucleic acid detection is expected to be a gold standard for noninvasive diagnosis of the new coronavirus all the time due to the core value and methodological advantages of pathogen detection in infection diagnosis and under further improvement of multiple ways.
Notomi et al, a novel molecular detection technique for isothermal amplification of specific fragments of nucleic acids in vitro in 2000, Loop-mediated isothermal amplification (LAMP), was developed. The technology utilizes 2 pairs of special primers to identify 6 sections of a target gene, DNA polymerase with strand displacement activity circularly generates a circular single-strand structure at the joint of the primers at two ends of a template under a constant temperature condition (60-65 ℃), and can carry out efficient and specific strand displacement amplification reaction on a target gene within 1 hour. Compared with the conventional PCR, the method does not need the processes of repeated temperature change, electrophoretic interpretation and the like, is higher than the PCR technology in technical indexes such as sensitivity, specificity, detection range and the like, can realize on-site rapid detection without depending on a special instrument, and has far lower cost than the PCR method.
Therefore, the LAMP technology is applied to virus detection, related diseases can be accurately and efficiently diagnosed, and research on the novel coronavirus is facilitated. The technology is further improved on the basis of the loop-mediated isothermal amplification method, the steps of sample collection, extraction and purification and the amplification efficiency are simplified, and the method has important significance for protecting the health of China and even people all over the world.
SUMMERY OF THE UTILITY MODEL
In order to solve the problem, the utility model provides a quick nucleic acid detection system. The visual LAMP reaction is utilized to detect the virus nucleic acid, and the sample collection, the nucleic acid extraction, the reaction and the observation are integrated, so that the operation is simple, and the application prospect is good.
The utility model provides a quick nucleic acid detecting device, nucleic acid detecting device includes: the device comprises a body part with an accommodating space and a sealing structure which is arranged on the body part and can be isolated and communicated, wherein the sealing structure divides the accommodating space into a first cavity for collecting a sample, a second cavity for accommodating a lysate and a third cavity for carrying out a nucleic acid amplification reaction.
In one embodiment, the second cavity is disposed between the first cavity and the third cavity, and the sealing structure includes a first sealing structure capable of communicating and separating the first cavity from the second cavity and a second sealing structure capable of communicating and separating the second cavity from the third cavity.
In one embodiment, the sealing structure is formed by any one or more of adhesive bonding, snap-fit engagement, film attachment, and external clamping.
In one embodiment, the sealing structure is a membrane attached to the body portion.
In one embodiment, at least one puncture structure for puncturing the film to realize communication between the cavities is arranged in the device.
In one embodiment, the piercing structure includes a connecting portion secured to the device and a tip extending from the connecting portion, the tip being movable along a plane to a piercing position of the membrane.
In one embodiment, the piercing structure is a planar structure substantially perpendicular to the film, the connecting portion is fixed to an edge of the body portion of the flexible structure, and the tip is an acute-angled structure extending from the connecting portion toward the inside of the body portion.
In one embodiment, the connecting portion is fixed to the middle of the body portion, and the tip is rotatable about the connecting portion to a piercing position of the membrane.
In one embodiment, the piercing structure is disposed within the second and/or third cavities.
In one embodiment, the sealing structure is provided with sealing lines on both sides thereof, the sealing lines extending to the edge of the body portion, and the sealing lines apply a force separating the sealing structure toward both sides by an external force.
In one embodiment, the sealing structure is provided with handles on both sides for applying the separation force.
In one embodiment, the nucleic acid amplification reagent is a dry reagent, the third cavity comprises a first sub-cavity for accommodating the dry reagent and a second sub-cavity for accommodating a desolvation, and a third sealing structure capable of communicating or blocking is arranged between the first sub-cavity and the second sub-cavity.
In one embodiment, the third sealing structure wraps the first sub-cavity, the first sub-cavity is arranged in the second sub-cavity, the third sealing structure is made of waterproof fragile materials, the dry reagent can be melted into the desolvation under the damage of external force, and the third sealing structure is preferably tinfoil paper.
In one embodiment, the second sub-cavity is located above the first sub-cavity, and the third sealing structure releases the double solvent to flow into the first sub-cavity to dissolve the dry reagent after being damaged by external force.
In one embodiment, the second and/or third sealing structure comprises a low melting point material layer, preferably a paraffin layer, which is meltable under water bath heating.
In one embodiment, the opening of the body portion is provided with a tear-off opening and/or a hidden button for opening or closing the first cavity to complete the collection of the sample.
In one embodiment, the first cavity is a tapered structure, the inner diameter of the first cavity gradually decreases from the opening of the body part to the first sealing structure, and an indicator line for the sample collection amount is arranged on the first cavity.
In one embodiment, the first sealing structure is provided with at least one through hole penetrating through the upper and lower surfaces of the first sealing structure, the diameter of the through hole is smaller than the length of the through hole, and the sample in the first cavity flows into the second cavity through the through hole under the action of external air pressure.
In one embodiment, the through holes are uniformly elongated micropores; or
The through holes are tapered micropores with the pore diameter from top to bottom from large to small.
In one embodiment, a one-way air valve is arranged on the first cavity, and after the first cavity is sealed, the one-way air valve can ventilate the first cavity, so that the sample is promoted to enter the second cavity through the through hole.
In one embodiment, the sealing structure includes a clamping mechanism separable from the body portion, the clamping mechanism clamping the body portion to achieve cavity isolation.
In one embodiment, the device is made of transparent or semitransparent plastic materials; or
The body part is made of opaque materials for shading light, and transparent observation windows are arranged at the corresponding positions of the first cavity and the third cavity.
In one embodiment, the body portion is provided with a fixing hole or a fixing member for fixing with the outside.
The utility model also provides a quick nucleic acid detecting system, it includes as above nucleic acid detecting device and be used for carrying out thermostatic control's temperature controller to the reaction observation part.
The utility model also provides a quick nucleic acid detecting device and system, the integration sets up, collects sample collection, nucleic acid extraction, reaction and observes as an organic whole. Need not medical personnel's assistance and can take a sample by oneself to need not the professional operation, only need mix the reaction reagent of sample and system, can accomplish nucleic acid detection and observe the testing result. The detection system is simple to operate, low in manufacturing cost and simple in preparation process, and the used main components are dry powder, so that the storage period is long and the stability of the reagent is good; therefore, the whole system has extremely good application prospect.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 3 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 4 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 5 is an enlarged view of portion A of FIG. 4 in one embodiment;
FIG. 6 is an enlarged view of portion A of FIG. 4 in another embodiment;
FIG. 7 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 8 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 9 is a schematic side view of the structure of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 10 is an enlarged view of the internal structure of portion B of FIGS. 4 and 7 in one embodiment;
FIG. 11 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 12 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 13 is a schematic structural diagram of a rapid nucleic acid detection system according to an embodiment of the present invention;
FIG. 14 is a schematic diagram of the external structure of a rapid nucleic acid detection system according to an embodiment of the present invention;
the reference numbers are as follows:
a sample collection portion-1; a first chamber-10; an opening-101; an indicator line-102; a fixed hole-103; sample pre-treatment section-2; a second cavity-20; reaction observing part-3; a third cavity-30; a first subchamber-301; a second subcavity-302; a third sub-cavity-33; a fourth subchamber-34; a seal structure-4; a first seal structure-41; a second seal structure-42; third seal structure-43; a puncture structure 5; a connecting part-501; a tip-502; a clamping mechanism-6.
Detailed Description
In order to make the technical solutions in the present application better understood by those skilled in the art, the present invention will be further described with reference to the following examples, and it should be apparent that the described examples are only a part of the examples of the present application, and not all examples. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
Definition of
The following definitions apply herein unless otherwise indicated.
The terms "comprising," "including," and "having" (and variations thereof) are used interchangeably to mean including, but not necessarily limited to, and are open-ended terms that are not intended to exclude additional unrecited elements or method steps.
Terms such as "first," "second," and "third" are used to distinguish or identify various members of a concept such as a group, and are not intended to represent sequential or numerical limitations.
Sample preparation: "sample", "sample" are used interchangeably to refer to a compound, composition, and/or mixture of interest from any suitable source. A sample is a general object of interest for a test that analyzes an aspect of the sample, such as an aspect associated with at least one analyte that may be present in the sample. The sample may be analyzed for its natural state at the time of collection and/or for an altered state, e.g., caused by: storing, preserving, extracting, lysing, diluting, concentrating (concentrating), purifying, filtering, mixing with one or more reagents, pre-amplifying (e.g., target enrichment by performing limited cycles of PCR on the sample prior to PCR (e.g., < 15)), removing amplicons (e.g., treating with uracil-d-glycosylase (UDG) prior to PCR to eliminate any carryover contamination by previously generated amplicons (i.e., amplicons can be digested with UDG because it was generated with dUTP instead of dTTP)), partitioning, or any combination thereof, and the like. Clinical samples may include nasopharyngeal washes, blood, plasma, acellular plasma, buffy coat, saliva, urine, stool, sputum, mucus, wound swabs, tissue biopsies, milk, fluid aspirates, swabs (e.g., nasopharyngeal swabs), and/or tissues, and the like. The utility model discloses a sample mainly indicates clinical sample.
Reagent: compounds, compositions of compounds and/or compositions that are combined with a sample to perform a particular test on the sample. The reagent may be a target-specific reagent, which is any reagent composition that makes a particular target or analyte in the assay specific for detection. Alternatively, the reagent may comprise a chemical reactant and/or a binding partner for the test. The reagents may for example include: at least one nucleic acid, protein (e.g., enzyme), cell, virus, organelle, macromolecular assembly, potential drug, lipid, carbohydrate, inorganic substance, or any combination thereof, and can be an aqueous composition, and the like. In exemplary embodiments, the reagents may be amplification reagents, which may include at least one primer or at least one pair of primers for amplifying a nucleic acid target, at least one probe and/or dye that enables detection of amplification, a polymerase, nucleotides (dntps and/or NTPs), divalent magnesium ions, potassium chloride, buffers, or any combination thereof, and the like.
Nucleic acid (A): a compound comprising a nucleotide monomer chain. A nucleic acid can be single-stranded or double-stranded (i.e., form base pairs with another nucleic acid), and the like. The nucleic acid strand may comprise any suitable number of monomers, such as at least about ten or one hundred, and the like. Generally, nucleic acid strands have a length corresponding to their origin, synthetic nucleic acids (e.g., primers and probes) are generally shorter, while biologically/enzymatically produced nucleic acids (e.g., nucleic acid analytes) are generally longer. The nucleic acid used by the utility model is limited to the nucleic acid of the novel coronavirus, and the novel coronavirus SARS-CoV-2 is a positive-sense single-stranded RNA virus; thus, the nucleic acid detection refers to RNA nucleic acid detection.
The sequence of the nucleic acid is defined by the order in which the nucleobases are arranged along the backbone. This sequence generally determines the ability of a nucleic acid to bind specifically to a partner strand (or form an intramolecular duplex) through hydrogen bonding. In particular, adenine pairs with thymine (or uracil), and guanine pairs with cytosine. Nucleic acids that can bind to another nucleic acid in an antiparallel manner by forming a contiguous string of such base pairs with the other nucleic acid are said to be "complementary".
Amplification: the reaction of replication to form multiple copies of at least one segment of the template molecule is repeated over time. Amplification can produce an exponential or linear increase in copy number as amplification proceeds. Typical amplification produces over a 1000-fold increase in copy number and/or signal.
"reverse transcription" refers to the formation of complementary DNA from an RNA template, such as any suitable transcription product. Reverse transcription can be performed with enzymes that catalyze this process. Exemplary enzymes that may be suitable include reverse transcriptases (e.g., MLV-RT, Tth polymerase, AMV-RT, HIV-RT, etc.). The enzyme may also be capable of synthesizing DNA using the DNA as a template, and in some cases, may be capable of amplifying a target sequence. The enzyme may be sufficiently thermostable to allow heating of the partition to open the partition without destroying reverse transcription activity of the enzyme. Reverse transcription and reverse transcription are used interchangeably.
The enzyme with strand displacement activity can perform strand displacement and amplification at a constant temperature, and can include, but is not limited to, Bst polymerase (such as Bst polymerase 1.0 and Bst polymerase 2.0), Gsp polymerase, and other DNA polymerases with similar high strand displacement activity. The magnesium salt may be magnesium sulfate or magnesium chloride. The magnesium salt is primarily used to catalyze the activity of the strand-displacement DNA polymerase.
In the present invention, the enzyme having strand displacement activity and the enzyme having reverse transcription activity may be the same enzyme, for example, Bst polymerase 4.0.
Primer: a nucleic acid capable of and/or for use in priming replication of a nucleic acid template. Thus, a primer is a shorter nucleic acid that is complementary to a longer template. During replication, the primer is extended based on the template sequence to produce a longer nucleic acid, which is a complementary copy of the template. The primer may be DNA, RNA, analogs thereof (i.e., artificial nucleic acids), or any combination thereof. The primer can be of any suitable length, such as at least about 10, 15, 20, or 30 nucleotides. Exemplary primers are chemically synthesized. The primers may be provided in the form of at least one pair of primers for amplifying at least one nucleic acid target. The pair of primers may be a sense primer and an antisense primer that together define opposite ends of the resulting amplicon (and thus define the length of the resulting amplicon).
Unless otherwise indicated, all chemical reagents referred to in the following description are commercially available reagents.
The dNTPs may be dNTPs conventional in the art, including dATP, dGTP, dTTP and dCTP.
"reconstitution liquid", "reconstitution solvent" are used interchangeably to mean reconstituting a dry powder nucleic acid amplification reagent into an aqueous solution reagent.
Example 1
As shown in fig. 1, the embodiment of the present invention provides a rapid nucleic acid detecting system, which includes:
a sample collection part 1 having a first cavity 10 for collecting a sample;
a reaction observing section 3 having a third chamber 30 for storing a nucleic acid amplification reagent, for performing a nucleic acid amplification reaction and observing a change before and after amplification; the nucleic acid amplification reagent comprises a primer composition, Bst polymerase 4.0, nucleic acid dye HNB and dNTP;
in one embodiment, the primer composition comprises a primer group corresponding to at least one of primer group SEQ ID NO 7-SEQ ID NO 12 corresponding to SARS-CoV-2 specific gene ORF1ab, and primer group corresponding to at least one of primer group SEQ ID NO 13-SEQ ID NO 18 corresponding to SARS-CoV-2 specific gene S; the nucleic acid dye is selected from cresol red, neutral red, 3-nitrophenol, m-cresol purple, naphthol peptide or methylene blue, erythrosine, allura red AC, tartrazine, sunset yellow FCF, indigo carmine, betaine, chlorophyll, caramel color, butterfly pea, vanilla, ultramarine, malachite blue, calcein Green, hydroxynaphthol blue, SYBR Green I, Eva Green, cobalt blue or phthalocyanine.
In one embodiment, the primer composition further comprises primer set 3 corresponding to 18s rRNA;
the primer group 3 is SEQ ID NO 1-4 or SEQ ID NO 1-6.
Further, the nucleic acid detection system further comprises:
a sample pretreatment section 2 having a second chamber 20 for containing a lysis solution. As shown in fig. 1, the sample pretreatment unit 2 is disposed between the sample collection unit 1 and the reaction observation unit 3, and the second chamber 20 and the first chamber 10 and the third chamber 30 are connected in a communication or blocking manner.
The sample pretreatment part 2 is used for pretreatment such as lysis and dilution of a sample, and a pretreatment solution such as a lysis solution is contained in the second cavity 20; the lysis solution is selected from deionized water, normal saline, cell lysis solution and the like. Preferably, the lysis solution further comprises protease and DNase.
When the sample is collected in a sufficient amount, the first cavity 10 and the second cavity 20 are communicated; fully mixing the sample and the pretreatment solution, breaking the joint between the second cavity 20 and the third cavity 30 by using external acting force such as pulling force traction and a sterile wood stick to communicate the sample and the third cavity, fully and uniformly mixing the pretreated sample and a nucleic acid amplification reagent in the third cavity, and carrying out constant temperature control on a reaction observation part by using a water bath, a constant temperature box and the like to start loop-mediated constant temperature amplification; after reacting for 45min, directly observing the color change before and after the reaction by naked eyes to obtain a detection result.
In a specific embodiment, when the nucleic acid amplification reagent is a dry reagent, as shown in fig. 2, the third cavity 30 has a first sub-cavity 301 and a second sub-cavity 302, the first sub-cavity 301 is for accommodating the dry reagent, and the second sub-cavity 302 is for accommodating the re-dissolving agent; the dry reagent may be released from the first sub-chamber and mixed with the re-solvent, or the re-solvent may be released from the second sub-chamber and mixed with the dry reagent, under the action of an external force, or when a specific condition is achieved.
In another embodiment, as shown in figure 3, the nucleic acid amplification reagents of the third and fourth sub-chambers 33, 34 are different for amplifying different target nucleic acid sequences. The primer composition in the nucleic acid amplification reagent in the third subcavity 33 is a primer group SEQ ID NO. 1-SEQ ID NO. 6 corresponding to 18s rRNA; the primer composition in the nucleic acid amplification reagent in the fourth subcavity 34 does not contain the primer sets SEQ ID NO. 1-SEQ ID NO. 6 corresponding to 18s rRNA. Wherein, the primer group corresponding to 18s rRNA is used as an internal reference for removing systematic errors.
In one embodiment, the nucleic acid detecting system further comprises a temperature controller for controlling the reaction observing section at a constant temperature, and the temperature controller is provided independently and is not connected to other structures of the nucleic acid detecting system. The temperature controller is selected from a temperature controller which can provide the temperature of 60-65 ℃ such as a water bath kettle, a thermostat and the like.
Example 2
The present embodiment provides a nucleic acid rapid detection apparatus implementing the rapid nucleic acid detection system of the above embodiment:
as shown in FIG. 4, the rapid nucleic acid detecting device of this embodiment includes a main body portion having an opening 101 at the top, and in this embodiment, the main body portion is a flexible bag-shaped structure and includes a first inner surface and a second inner surface that are oppositely disposed, and the side edges and the bottom of the first inner surface and the second inner surface are combined together by a hot pressing method to form an accommodating space that is relatively independent from the external environment.
The rapid nucleic acid detecting device further comprises a sealing structure 4 which divides the containing space of the device into a plurality of areas and forms a plurality of cavities with different functions. Taking the rapid nucleic acid detecting apparatus of the present embodiment as an example, the apparatus has two sealing structures 4, namely, the first sealing structure 41 and the second sealing structure 42, the accommodating space of the apparatus is divided into three parts, namely, the first chamber 10, the second chamber 20 and the third chamber 30, and the main body part is divided into the sample collecting part 1, the sample pretreating part 2 and the reaction observing part 3.
The sealing structure 4 is a separable structure formed by combining the first inner surface and the second inner surface of the body part, and the first inner surface and the second inner surface are separated under the action of external force to realize the communication of two adjacent cavities.
Specifically, the rapid nucleic acid detection device of the present embodiment includes:
a sample collection section 1 for collecting a sample at the opening, a sample pretreatment section 2 for containing a lysate, and a reaction observation section 3 for performing a nucleic acid amplification reaction and observing a change before and after amplification.
The sample collection portion 1 is located at an opening 101 of the device, and includes the opening 101 and the first chamber 10 communicating with the opening 101. First cavity 10 is the back taper structure, from opening 101 to first seal structure 41, and the internal diameter of first cavity 10 reduces gradually, like this, is favorable to the gathering and the drainage of the sample of receiving.
In one embodiment, the opening 101 is provided with a hidden button or a sealing strip for sealing the first chamber 10 and the entire internal space, and the sample is sealed after collecting the sample such as saliva, so as to effectively prevent the sample from polluting the environment.
Preferably, the opening 101 is further provided with a tear-off opening, which is convenient for a user to open the device to complete sample collection.
In one embodiment, the sample collection portion 1 is provided with a sample indication line for measuring the sample collection volume, such as by observing the total amount of liquid after the saliva sample is directly spit into the sample collection portion to determine whether the proper amount of liquid has been collected.
The sample pretreatment section 2 includes a second chamber 20 containing a lysis solution, and a first sealing structure 41 for separating the first chamber 10 from the second chamber 20. In the present embodiment, the first sealing structure 41 is a hard structure having a layer shape, and at least one through hole 4101 penetrating the upper and lower surfaces is provided in the first sealing structure 41, and preferably, the hole diameter of the through hole 4101 is smaller than the hole length of the through hole 4101.
In one embodiment, as shown in fig. 5, the through holes 4101 are elongated micro holes with uniform inner diameter; in a normal state, since the air pressure between the first chamber 10 and the second chamber 20 is balanced, the sample collected in the first chamber 10 cannot naturally pass through the through hole, and the first chamber 10 and the second chamber 20 are in an isolated state. After the sample collection amount reaches the position of the indicating line, the air pressure of the first chamber 10 is increased by blowing or ventilating the first chamber 10, so that the sample is forced to enter the second chamber 20 through the through hole.
In another embodiment, as shown in fig. 6, in order to prevent the liquid in the second chamber 20 from flowing backward into the first chamber during shaking, water bath, etc., the through holes 4101' are provided as tapered pores with a diameter from large to small from top to bottom.
In order to facilitate the user to blow or ventilate into the first cavity, a one-way air valve is further arranged on the sample collecting part 1. The one-way air valve can be further connected with an air blowing hole, so that a user can conveniently ventilate the closed first cavity when using the one-way air valve, and a sample is promoted to enter the second cavity.
The reaction observing section 3 includes a third chamber 30 for performing a nucleic acid amplification reaction and observing changes before and after amplification. The nucleic acid amplification reagent comprises a primer composition, Bst polymerase 4.0, nucleic acid dye HNB and dNTP.
In the present embodiment, the nucleic acid amplification reagent is a liquid reagent. A second seal 42 is disposed between the second cavity 20 and the third cavity 30. In the present embodiment, the second seal structure 42 is a paraffin layer or a layered structure containing at least paraffin.
In alternative embodiments, the sealing structure may take other forms, such as adhesive bonding, snap-fit engagement, film attachment, and external clamping, as long as communication under specific conditions is achieved.
After a proper amount of sample such as saliva is collected by the sample collecting part 1, the sample enters the second cavity 20 through ventilation, and is fully mixed with the lysate, and then the device is placed into a water bath, a thermostat and the like to carry out thermostatic control on the device. The paraffin wax has an amorphous structure, and the melting point is not constant, and is generally 49-51 ℃. In the process of carrying out constant temperature control on the sample at 60-65 ℃, the paraffin layer melts after reaching the melting point along with the rise of the external temperature, and the second sealing structure 42 is damaged, so that the second cavity 20 is communicated with the third cavity 30. Further, the whole device can be shaken to fully mix the pre-treated sample with the nucleic acid amplification reagent in the third cavity 30, and constant temperature control is continuously performed to start loop-mediated constant temperature amplification; after reacting for 45min, directly observing the color change before and after the reaction by naked eyes to obtain a detection result.
The rapid nucleic acid detection device is prepared by adopting a flexible material which does not influence nucleic acid amplification and does not react with a clinical sample, preferably adopts a transparent or semitransparent plastic material, in other rotatable embodiments, an opaque material can be adopted, and the reaction observation part 3 and the sample collection part 1 are provided with transparent observation windows so as to observe the condition of the device conveniently.
Specifically, the flexible material can be selected from, but not limited to, transparent or translucent PET, PDMS, POE, EVA, EPDM, PPP, silicone, PC, PPSU, and the like.
Further, still be provided with on the quick nucleic acid testing device and be used for with external fixed complex fixed orifices 103 or mounting, when placing the device and carry out thermostatic control in the water bath, can hang the device on external environment's couple through fixed orifices 103, or hang the device on external environment's fixed part through the couple.
Example 3
As shown in FIG. 7, this example provides a rapid nucleic acid detecting apparatus, which is basically the same as example 2 except that in this embodiment, the nucleic acid amplification reagent is a dry reagent.
The rapid nucleic acid detecting apparatus includes a sample collecting part 1 having a first chamber 10 for collecting a sample, a sample pretreating part 2 having a second chamber 20 for accommodating a lysate, and a reaction observing part 3 having a third chamber 30 for performing a nucleic acid amplification reaction and observing a change before and after amplification.
The third cavity 30 includes a first sub-cavity 301 for accommodating a dry reagent and a second sub-cavity 302 for accommodating a desolvation, and a third sealing structure 43 capable of communicating or blocking is disposed between the first sub-cavity 301 and the second sub-cavity.
In this embodiment, the third sealing structure 43 encapsulates the dry reagent to form a first sub-chamber 301, the first sub-chamber 301 being disposed within a second sub-chamber 302. The third sealing structure is made of waterproof fragile materials, can be damaged under the action of external force and releases the dry reagent in the third sealing structure, and the dry reagent is mixed with the liquid such as the double-solvent in the second sub-cavity 302 to form the nucleic acid amplification reagent. Preferably, the third sealing structure is made of a tin foil material.
In the use process, a user only needs to crush and pinch the first sub-cavity through fingers to break the third sealing structure 43 for sealing the dry reagent, and the redissolution of the dry reagent and the preparation of the nucleic acid amplification solution are completed.
It should be noted that the operation of destroying the third sealing structure 43 to complete the reconstitution of the dry reagent may be performed before the sample pretreatment is completed, or may be completed after the sample pretreatment; preferably, after the sample is pretreated and the sample and the pretreatment reagent are mixed, the redissolution of the dry reagent can be carried out, and then the constant temperature control can be carried out.
In alternative embodiments, the third sealing structure 43 may also be a paraffin layer, and during the constant temperature control process, the third sealing structure 43 is broken to release the dry reagent, and then the dry reagent is dissolved into the mixture of the double solvents to form the sample amplification solution.
In another embodiment, as shown in fig. 8, the second sub-chamber 302 is located above the first sub-chamber 301, the third sealing structure is a paraffin layer, after thermostatic control in a water bath, the paraffin layer is destroyed, and the second sub-chamber 302 releases the double solvent to flow into the first sub-chamber to dissolve the dry reagent located at the bottom of the first sub-chamber to form the nucleic acid amplification solution.
In another embodiment, as shown in fig. 9, the first and second seal structures 41 and 42 may be separable structures formed by combining the first inner surface and the second inner surface of the body, and the first inner surface and the second inner surface are separated under the action of external force to communicate with two adjacent cavities.
Specifically, the first sealing structure 41 and the second sealing structure 42 are respectively provided with hidden buttons 401 which can be matched and combined on the first inner surface and the second inner surface, and the hidden buttons are combined together by pressing two sides to realize the sealing of two adjacent cavities. In the use process, the hidden button is separated by external pulling, so that the communication between two adjacent cavities is realized.
Further, as shown in fig. 10, the first seal structure 41 or the second seal structure 42 further has a seal line 402 respectively connected to the first inner surface and the second inner surface, and the seal line 402 extends to the edge of the body portion and forms a protrusion at the device edge. When the first sealing structure 41 or the second sealing structure 42 needs to be communicated, the sealing lines only need to be pulled to two sides, external force can be applied to the first inner surface and the second inner surface, the hidden buckle is opened, and communication of adjacent cavities is achieved.
In another embodiment, a pull handle for separating the first inner surface from the second inner surface may be provided on the first seal structure 41 or the second seal structure 42, where the pull handle may be a hard structure or a soft structure, and the hidden button is opened by pulling the pull handle to both sides, so as to achieve communication between adjacent cavities.
It should be noted that, in the present embodiment, the first sealing structure 41 and the second sealing structure 42 are taken as examples to describe specific forms of the sealing structures, and in fact, the sealing structures are not limited to the first sealing structure 41 and the second sealing structure 42, and in fact, the third sealing structure 43 may also adopt corresponding forms, which are not described herein again.
In another embodiment, the sealing structure 4 may also be an intermediate layer disposed between the first inner surface and the second inner surface, and the intermediate layer may be a glue layer formed by bonding with an adhesive, a film or a mesh disposed between the first inner surface and the second inner surface, or a transition connection of the same material as the body portion disposed between the first inner surface and the second inner surface. The middle layer is designed to be easily punctured and torn by adjusting the thickness of the material and selecting the material; under the action of external force, the sealing structure 4 is broken, the first inner surface and the second inner surface are separated, and the two adjacent cavities are communicated. Preferably, the middle layer is a film which is broken when being pulled and torn by external tension or poked by a sterile wooden stick, and the two adjacent cavities are communicated.
Further, as shown in fig. 11, in one embodiment, in order to facilitate the destruction of the intermediate layer and to quickly achieve the communication between the adjacent cavities, a puncturing structure for puncturing the intermediate layer may be further provided in the device.
In one embodiment, the piercing structure 5 is a planar structure substantially perpendicular to the membrane, and the piercing structure 5 includes a connecting portion connected to the body portion and at least one tip that is movable with the flexible structure along the plane to a piercing position of the first sealing structure 41 or the second sealing structure 42 to pierce and break the intermediate layer and communicate the adjacent cavities. In this embodiment, the puncturing structure is a planar triangular structure, the connecting portion is fixed to an edge of the main body portion of the flexible structure, and the tip is an acute-angle structure extending from the connecting portion to an inside of the main body portion and used for puncturing the intermediate layer to achieve cavity communication.
The puncture arrangement 5 may be arranged in the second cavity 20, or in the third cavity 30, preferably in the second cavity. In this way, only one puncture structure is provided, and puncture by the first seal structure 41 and puncture by the second seal structure 42 can be achieved. In other alternative embodiments, piercing structures may also be provided in both the second lumen 20 and the third lumen 30.
In some cases, as shown in FIG. 12, the third seal 43 may also communicate using a piercing structure. For example, when the second sub-cavity 302 containing the desolvation in the third cavity 30 is arranged above the first sub-cavity 301, it is preferable to provide a puncturing structure 5 in the second sub-cavity 302, and the puncturing structure 5 can achieve puncturing of the second sealing structure and can also puncture the third sealing structure 43, so that the desolvation enters the first sub-cavity at the bottom to dissolve the dry reagent therein.
In another embodiment, as shown in FIG. 13, the piercing structure is a rotatable needle-like structure or a knife-like structure, the connecting portion 501 is disposed at a central position of the body portion, and the tip 502 is rotatable about the connecting portion to a piercing position of the membrane. In the non-piercing stage, the tip 502 is in a horizontal position. Preferably, the horizontal position is provided with a limit structure of the connecting part, so that the puncture structure can keep the horizontal position unmovable without puncturing, and unnecessary damage to the body part and the system is avoided. When puncture is needed, the tip of the puncture structure moves to the puncture position of the first sealing structure or the second sealing structure through rotation of the connecting part, puncture damage is carried out on the middle layer, and communication of adjacent cavities is achieved.
In another particular embodiment, as shown in fig. 14, the sealing structure 4 includes a clamping mechanism 6 that is separable from the body portion. When the cavity needs to be isolated, the clamping mechanism 6 is fixed on the outer side of the body part, and the accommodating space of the device is divided into a plurality of cavities, which correspond to the first cavity 10, the second cavity 20 and the third cavity 30 of the system respectively. In the process of using by the user, when the collection of the sample is completed and the lysis is required, only the clamping mechanism between the first cavity 10 and the second cavity 20 needs to be opened. Similarly, when the lysis is completed, the pre-treated sample and the amplification reagent can be fused by opening the clamping mechanism between the second cavity 20 and the third cavity 30.
Further, the clamping mechanism 6 comprises a clamping body 61 and a fastener 62 slidable along the clamping body, and after the clamping body 61 is clamped to the outside of the device, the fastener 62 further presses the clamping mechanism to seal the inner cavity of the device.
The clamping mechanism 6 of the present embodiment may be used only for separating the first cavity 10 from the second cavity 20, or may be used only for separating the second cavity 20 from the third cavity 30. Preferably, two clamping mechanisms 6 are provided, while achieving separation of the three chambers. The structure has low cost and is convenient for users to operate. Furthermore, when two separate sub-chambers are provided within the third chamber 30, the clamping mechanism 6 may also be used to separate the first sub-chamber 301 from the second sub-chamber 302. In the rapid nucleic acid detection system of the embodiment, in the production process, only the reagent of the cavity at the bottommost layer needs to be filled first, and the reagent of the upper layer is filled after the clamping mechanism is installed for sealing.
The sealing structure of the invention aims to realize the communication and the obstruction between two adjacent cavities, and the two adjacent cavities are in an obstruction state during transportation and preservation; when the device is used by a user, the communication between two adjacent cavities can be realized by external force damage, the above embodiments are merely examples, and the communication or the blockage is any form of communication or blockage, including but not limited to adhesion, thin film connection, a communication valve, a paraffin layer, and the like, which are not exemplified herein.
Fig. 11 shows a preferred embodiment of the present invention, in which the first sealing structure 41 is a hard structure with a plurality of through holes, and the communication is realized by blowing or ventilating air by the user; the third sealing structure 43 is preferably made of a brittle material which can be damaged under the action of external force, and a user can knead and explode the first sub-cavity to redissolve the dry reagent in the using process; the second sealing structure 42 is preferably punctured by the puncturing structure 5 to realize the cavity communication or in a hidden button mode, and the cavity communication is realized by pulling and destroying the second sealing structure towards two sides by a user. The method has the advantages of low cost and simple user operation, and can realize quick cavity communication and facilitate the acceleration of detection speed.
As shown in fig. 8, for the embodiment of the present invention, the second sealing structure 42 and the third sealing structure 43 are paraffin layers, after the reagent is fully cracked, when the temperature control device such as the water bath is placed in the body part to perform the start-up loop-mediated isothermal amplification, the paraffin layers are melted, and the second sealing layer 42 and the third sealing layer 43 are connected simultaneously. The method has the advantages that the reaction environment of the system can be fully utilized, and the detection can be completed without other operations of a user.
The utility model also provides a quick nucleic acid detecting device and system, the integration sets up, collects sample collection, nucleic acid extraction, reaction and observes as an organic whole. Need not medical personnel's assistance and can take a sample by oneself to need not the professional operation, only need mix the reaction reagent of sample and system, can accomplish nucleic acid detection and observe the testing result. The detection system is simple to operate, low in manufacturing cost and simple in preparation process, and the used main components are dry powder, so that the storage period is long and the stability of the reagent is good; therefore, the whole system has extremely good application prospect.
It should be noted that reference in the specification to "an embodiment," "one embodiment," "some embodiments," or "other embodiments" means that a particular feature, structure, or characteristic described in connection with the embodiments is included in at least some embodiments, but not necessarily all embodiments. The embodiments in the present specification are described in a progressive manner, and the same or similar parts in the embodiments are referred to each other, and each embodiment focuses on the differences from the other embodiments. The present application is not limited to the particular steps and structures described above and shown in the drawings. Those skilled in the art may make various changes, modifications and additions or change the order between the steps after appreciating the spirit of the present application. Those skilled in the art will also recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.
Claims (23)
1. A rapid nucleic acid detecting apparatus, comprising: the device comprises a body part with an accommodating space and a sealing structure which is arranged on the body part and can be isolated and communicated, wherein the sealing structure divides the accommodating space into a first cavity for collecting a sample, a second cavity for accommodating a lysate and a third cavity for carrying out a nucleic acid amplification reaction;
the device is made of transparent or semitransparent plastic materials; or
The body part is made of opaque materials for shading light, and transparent observation windows are arranged at the corresponding positions of the first cavity and the third cavity.
2. The apparatus according to claim 1, wherein the second chamber is disposed between the first chamber and the third chamber, and the sealing structure comprises a first sealing structure capable of communicating and separating the first chamber from the second chamber, and a second sealing structure capable of communicating and separating the second chamber from the third chamber.
3. The apparatus for rapid nucleic acid detection according to claim 1, wherein the sealing structure is formed by any one or more of adhesive bonding, snap-fit engagement, film attachment, and external force clamping.
4. The apparatus according to claim 2, wherein the sealing structure is a membrane connected to the body, and at least one piercing structure is disposed in the apparatus for piercing the membrane to communicate the cavities.
5. The apparatus according to claim 4, wherein the piercing structure comprises a connecting portion fixed to the body portion and a tip extending from the connecting portion, the tip being movable along the plane to a piercing position of the thin film.
6. The apparatus for rapid nucleic acid detection according to claim 5, wherein the piercing structure is a planar structure substantially perpendicular to the film, the connecting portion is fixed to an edge of the main body portion of the flexible structure, and the tip is an acute-angled structure extending from the connecting portion toward an inside of the main body portion.
7. The apparatus for rapid nucleic acid detection according to claim 5, wherein the connecting portion is fixed to the middle portion of the body portion, and the tip is rotatable around the connecting portion to a piercing position of the membrane.
8. The rapid nucleic acid detection device of claim 4, wherein the puncture structure is disposed within the second cavity and/or the third cavity.
9. The apparatus for rapid nucleic acid detection according to claim 3, wherein the sealing structure is provided with sealing lines on both sides thereof, the sealing lines extending to the edge of the body portion, the sealing lines applying a force to the sealing structure separating to both sides under an external force.
10. The apparatus for rapid nucleic acid detection according to claim 3, wherein both sides of the sealing structure are provided with a handle for applying a separating force.
11. The rapid nucleic acid detection device according to claim 2, wherein the nucleic acid amplification reagent is a dry reagent, the third cavity comprises a first sub-cavity for accommodating the dry reagent and a second sub-cavity for accommodating a desolvation, and a third sealing structure capable of communicating or blocking is arranged between the first sub-cavity and the second sub-cavity.
12. The apparatus according to claim 11, wherein the third sealing structure encloses the first sub-cavity, the first sub-cavity is disposed in the second sub-cavity, and the third sealing structure is made of a waterproof brittle material, and the dry reagent can be dissolved into the desolvation under the damage of an external force.
13. The apparatus for rapid nucleic acid detection according to claim 12, wherein the third sealing structure is a foil.
14. The rapid nucleic acid detecting device according to claim 11, wherein the second sealing structure and/or the third sealing structure comprises a low melting point material layer that is meltable under water bath heating.
15. The apparatus for rapid nucleic acid detection according to claim 14, wherein the low-melting-point material layer is a paraffin layer.
16. The rapid nucleic acid detecting device according to claim 2, wherein the opening of the body portion is provided with a tear-off opening and/or a hidden button for opening or closing the first cavity to complete the collection of the sample.
17. The apparatus according to claim 16, wherein the first chamber has a tapered structure, and an inner diameter of the first chamber gradually decreases from the opening of the body to the first sealing structure, and the first chamber is provided with a sample collection amount indicator.
18. The apparatus for rapid nucleic acid detection according to claim 17, wherein the first sealing structure is provided with at least one through hole penetrating upper and lower surfaces of the first sealing structure, the through hole has a smaller diameter than a hole length of the through hole, and the sample in the first chamber flows into the second chamber through the through hole under the action of the external air pressure.
19. The apparatus for rapid nucleic acid detection according to claim 18, wherein the through-holes are elongated micro-holes with uniform inner diameter; or
The through holes are tapered micropores with the pore diameter from top to bottom from large to small.
20. The apparatus according to claim 18, wherein a one-way air valve is disposed on the first chamber, and after the first chamber is sealed, the one-way air valve can vent the first chamber to promote the sample to enter the second chamber through the through hole.
21. The apparatus according to claim 1, wherein the sealing structure comprises a clamping mechanism separable from the body portion, the clamping mechanism clamping the body portion to achieve cavity isolation.
22. The apparatus for rapid nucleic acid detection according to claim 1, wherein the body is provided with a fixing hole or a fixing member for fitting and fixing with the outside.
23. A rapid nucleic acid detecting system comprising the rapid nucleic acid detecting apparatus according to any one of claims 1 to 22 and a temperature controller for controlling the reaction observing section at a constant temperature.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202021381274.7U CN213951203U (en) | 2020-07-14 | 2020-07-14 | Rapid nucleic acid detection system and device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202021381274.7U CN213951203U (en) | 2020-07-14 | 2020-07-14 | Rapid nucleic acid detection system and device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN213951203U true CN213951203U (en) | 2021-08-13 |
Family
ID=77192920
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202021381274.7U Expired - Fee Related CN213951203U (en) | 2020-07-14 | 2020-07-14 | Rapid nucleic acid detection system and device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN213951203U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023202246A1 (en) * | 2022-04-18 | 2023-10-26 | 杭州安旭生物科技股份有限公司 | Nucleic acid detection apparatus and nucleic acid detection method |
-
2020
- 2020-07-14 CN CN202021381274.7U patent/CN213951203U/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023202246A1 (en) * | 2022-04-18 | 2023-10-26 | 杭州安旭生物科技股份有限公司 | Nucleic acid detection apparatus and nucleic acid detection method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6153425A (en) | Self-contained device integrating nucleic acid extraction, amplification and detection | |
| US9315858B2 (en) | Apparatus for point-of-care detection of nucleic acid in a sample | |
| WO2004092342A2 (en) | Lateral flow system for nucleic acid detection | |
| CN105734168A (en) | Multiplex PCR detection kit for nucleic acids of twelve respiratory viruses | |
| KR20180080241A (en) | Sample preparation for difficult sample types | |
| US20180242896A1 (en) | Devices and methods for sample collection | |
| CN112029900A (en) | Rapid nucleic acid detection method and detection system for novel coronavirus | |
| WO2007100500A2 (en) | Sample processing | |
| Chu et al. | Recent advances and challenges of biosensing in point-of-care molecular diagnosis | |
| WO2011121454A2 (en) | Direct nucleic acid analysis | |
| Li et al. | Recent advancements in nucleic acid detection with microfluidic chip for molecular diagnostics | |
| CN213951203U (en) | Rapid nucleic acid detection system and device | |
| WO2007084567A3 (en) | Detection and discrimination of hepatitis c virus, human immunodeficiency virus type-1 and hepatitis b virus | |
| CN102337351A (en) | Typing detection kit for influenza virus | |
| US10590469B2 (en) | Methods for performing multiplexed real-time PCR in a self-contained nucleic acid analysis pouch | |
| CN105483293B (en) | Deadly infectious disease pathogen detection primer sets and kit | |
| CN107267668A (en) | Ring mediated isothermal amplification combination lateral flow test strips method detects SVCV | |
| CN111004719A (en) | Nucleic acid detection module, detection unit and detection system | |
| CN104962637A (en) | Kit and primer pair group for detecting drug-induced deafness susceptibility gene mutation site | |
| Bustin | The PCR revolution: basic technologies and applications | |
| CN112760210B (en) | Novel coronavirus integrated nucleic acid rapid detection card box | |
| WO2021229582A1 (en) | Device, systems, kits and methods for rapid and simple detection of pathogens | |
| CN103361438B (en) | A kind of template amplification and signal amplify the method combining detection nucleic acid | |
| CN113025474A (en) | Constant temperature nucleic acid amplification reaction tube and application thereof | |
| WO2009149115A1 (en) | Cartridge for conducting biochemical assays |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210813 |