CN104447959A - Polypeptide, detection device containing polypeptide and detection reagent kit provided with device - Google Patents
Polypeptide, detection device containing polypeptide and detection reagent kit provided with device Download PDFInfo
- Publication number
- CN104447959A CN104447959A CN201410848156.5A CN201410848156A CN104447959A CN 104447959 A CN104447959 A CN 104447959A CN 201410848156 A CN201410848156 A CN 201410848156A CN 104447959 A CN104447959 A CN 104447959A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- detection
- present
- detection means
- detection kit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 74
- 238000001514 detection method Methods 0.000 title claims abstract description 68
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 58
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 58
- 239000003153 chemical reaction reagent Substances 0.000 title abstract 3
- 238000003745 diagnosis Methods 0.000 claims abstract description 19
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical class O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 32
- 239000007787 solid Substances 0.000 claims description 25
- 238000002360 preparation method Methods 0.000 claims description 5
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 241001529459 Enterovirus A71 Species 0.000 abstract description 8
- 230000000694 effects Effects 0.000 abstract description 3
- 210000002966 serum Anatomy 0.000 description 24
- 239000000243 solution Substances 0.000 description 20
- 238000006243 chemical reaction Methods 0.000 description 15
- 239000000758 substrate Substances 0.000 description 15
- 238000005406 washing Methods 0.000 description 15
- 239000006210 lotion Substances 0.000 description 14
- 238000000034 method Methods 0.000 description 14
- -1 polypropylene Polymers 0.000 description 14
- 238000002493 microarray Methods 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 239000000463 material Substances 0.000 description 12
- 235000018102 proteins Nutrition 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 239000007788 liquid Substances 0.000 description 10
- 239000000203 mixture Substances 0.000 description 10
- 241000700605 Viruses Species 0.000 description 9
- 150000001413 amino acids Chemical group 0.000 description 9
- 238000002156 mixing Methods 0.000 description 9
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 239000011324 bead Substances 0.000 description 7
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 7
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 6
- 239000004205 dimethyl polysiloxane Substances 0.000 description 6
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 6
- 239000012898 sample dilution Substances 0.000 description 6
- 235000001014 amino acid Nutrition 0.000 description 5
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 5
- 239000003999 initiator Substances 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 229920001223 polyethylene glycol Polymers 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000005070 sampling Methods 0.000 description 4
- 239000000377 silicon dioxide Substances 0.000 description 4
- 229960001866 silicon dioxide Drugs 0.000 description 4
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 4
- 229920002554 vinyl polymer Polymers 0.000 description 4
- 108010016626 Dipeptides Proteins 0.000 description 3
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 102000003960 Ligases Human genes 0.000 description 3
- 108090000364 Ligases Proteins 0.000 description 3
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 229920002684 Sepharose Polymers 0.000 description 3
- 230000017531 blood circulation Effects 0.000 description 3
- 210000003169 central nervous system Anatomy 0.000 description 3
- 238000013016 damping Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 3
- 238000006116 polymerization reaction Methods 0.000 description 3
- 239000002243 precursor Substances 0.000 description 3
- 239000008213 purified water Substances 0.000 description 3
- 235000012239 silicon dioxide Nutrition 0.000 description 3
- 229920002379 silicone rubber Polymers 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 229920002818 (Hydroxyethyl)methacrylate Polymers 0.000 description 2
- CVEPFOUZABPRMK-UHFFFAOYSA-N 2-methylprop-2-enoic acid;styrene Chemical compound CC(=C)C(O)=O.C=CC1=CC=CC=C1 CVEPFOUZABPRMK-UHFFFAOYSA-N 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 208000020061 Hand, Foot and Mouth Disease Diseases 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 2
- 229910004298 SiO 2 Inorganic materials 0.000 description 2
- 206010058874 Viraemia Diseases 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000012790 confirmation Methods 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 230000005281 excited state Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000004816 latex Substances 0.000 description 2
- 229920000126 latex Polymers 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 208000030194 mouth disease Diseases 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000007170 pathology Effects 0.000 description 2
- 229920002223 polystyrene Polymers 0.000 description 2
- 238000010532 solid phase synthesis reaction Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- WGLQHUKCXBXUDV-UHFFFAOYSA-N 3-aminophthalic acid Chemical compound NC1=CC=CC(C(O)=O)=C1C(O)=O WGLQHUKCXBXUDV-UHFFFAOYSA-N 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 208000006740 Aseptic Meningitis Diseases 0.000 description 1
- SBHUBSDEZQFJHJ-CIUDSAMLSA-N Asp-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O SBHUBSDEZQFJHJ-CIUDSAMLSA-N 0.000 description 1
- IOXWDLNHXZOXQP-FXQIFTODSA-N Asp-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC(=O)O)N IOXWDLNHXZOXQP-FXQIFTODSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 208000014912 Central Nervous System Infections Diseases 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- QADHATDBZXHRCA-ACZMJKKPSA-N Cys-Gln-Asn Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CS)N QADHATDBZXHRCA-ACZMJKKPSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- SOEXCCGNHQBFPV-DLOVCJGASA-N Gln-Val-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(O)=O SOEXCCGNHQBFPV-DLOVCJGASA-N 0.000 description 1
- 208000025713 Hand-foot-and-mouth disease Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- WOBHKFSMXKNTIM-UHFFFAOYSA-N Hydroxyethyl methacrylate Chemical compound CC(=C)C(=O)OCCO WOBHKFSMXKNTIM-UHFFFAOYSA-N 0.000 description 1
- 206010061216 Infarction Diseases 0.000 description 1
- 244000207740 Lemna minor Species 0.000 description 1
- 235000006439 Lemna minor Nutrition 0.000 description 1
- 206010027201 Meningitis aseptic Diseases 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 208000000474 Poliomyelitis Diseases 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 235000001855 Portulaca oleracea Nutrition 0.000 description 1
- SGCZFWSQERRKBD-BQBZGAKWSA-N Pro-Asp-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 SGCZFWSQERRKBD-BQBZGAKWSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical class [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 Teflon® Polymers 0.000 description 1
- SBYQHZCMVSPQCS-RCWTZXSCSA-N Thr-Val-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(O)=O SBYQHZCMVSPQCS-RCWTZXSCSA-N 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 230000009435 amidation Effects 0.000 description 1
- 238000007112 amidation reaction Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000000133 brain stem Anatomy 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000002041 carbon nanotube Substances 0.000 description 1
- 229910021393 carbon nanotube Inorganic materials 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 208000015114 central nervous system disease Diseases 0.000 description 1
- 210000001638 cerebellum Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000024835 cytogamy Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 210000002451 diencephalon Anatomy 0.000 description 1
- 229910001882 dioxygen Inorganic materials 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 210000003038 endothelium Anatomy 0.000 description 1
- 238000012407 engineering method Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 230000005283 ground state Effects 0.000 description 1
- UQEAIHBTYFGYIE-UHFFFAOYSA-N hexamethyldisiloxane Polymers C[Si](C)(C)O[Si](C)(C)C UQEAIHBTYFGYIE-UHFFFAOYSA-N 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000005660 hydrophilic surface Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000007574 infarction Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000002198 insoluble material Substances 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000006011 modification reaction Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- HOGDNTQCSIKEEV-UHFFFAOYSA-N n'-hydroxybutanediamide Chemical compound NC(=O)CCC(=O)NO HOGDNTQCSIKEEV-UHFFFAOYSA-N 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000002887 neurotoxic effect Effects 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229920001778 nylon Polymers 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
- 210000000056 organ Anatomy 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000002940 repellent Effects 0.000 description 1
- 239000005871 repellent Substances 0.000 description 1
- 230000033458 reproduction Effects 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000004945 silicone rubber Substances 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000000278 spinal cord Anatomy 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000012876 topography Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000003156 vasculitic effect Effects 0.000 description 1
- 229910000859 α-Fe Inorganic materials 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention relates to polypeptide, a detection device containing the polypeptide and a detection reagent kit provided with the device. The polypeptide, the detection device containing the polypeptide and the detection reagent kit provided with the device are applied to clinical auxiliary diagnosis of an antibody IgG of the enterovirus 71, and a satisfying effect can be obtained.
Description
Technical field
The invention belongs to field of biological detection, the polypeptide that particularly can be used for the clinical assistant diagnosis of enterovirns type 71 IgG antibody, the detection kit of detection means comprising this polypeptide.
Background technology
Enterovirns type 71 (Human enterovirus 71), being called for short EV71, is cause Children (hand-foot and mouth disease; HFMD) one of main pathogens.Mankind's enteric virus71 type to suffer from from California first in 1969 and separates the infant faeces sample of central nervous system disease.
EV71 mainly causes hand foot mouth disease, also can cause the multiple nervous system disorderss such as the paralysis of aseptic meningitis, BBE and poliomyelitis sample.The two large common symptoms that hand foot mouth disease and central nervous system infection are EV71 infection and cause.Virus invades from pharyngeal or enteron aisle, breeds, and by locally discharging, now can cause local symptom in local mucous membrane or Lymphoid tissue.Then virus invades regional nodes again, and enter thus blood circulation cause first time viremia.Virus invades place's amount reproductions such as reticuloendothelium, deep layer lymphoglandula, liver, spleen, marrow through circulation of blood and enters blood circulation thus, causes second time viremia.Virus can enter each organ of whole body with blood flow, as places such as central nervous system, skin mucosa, hearts, breeds and causes pathology further.After susceptible person infects EV71, there is blood vessel transformation reactions and tissue inflammation pathology.When virus involves central nervous system, tissue inflammation comparatively neurotoxic effect is more strong, and central nervous system thin vessels endothelium is vulnerable to infringement most.Cytogamy, vasculitic become, thrombosis can cause ischemic and infarct.In the local organization of funiculi of spinal cord, brain stem, diencephalon, brain and cerebellum, except Neural invasion effect, also there are around blood vessel and parenchyma inflammation road virus 71 types widely very harmful, being badly in need of simple and efficient instrument accurately is at present realize the rapid detection to it.
Summary of the invention
The object of the present invention is to provide a peptide species, comprise the detection means of this polypeptide and comprise the detection kit of this detection means, they are useful to the clinical assistant diagnosis of enterovirns type 71 IgG antibody.The object of the invention is to provide this polypeptide to can be used for the purposes in the detection kit of enterovirns type 71 diagnosis in preparation.
That is, the present invention comprises following technical proposals:
1. a peptide species, its aminoacid sequence as shown in SEQ ID NO:1, that is: QVVTVMDDLCQNPDGKDMSL.
2. a detection means, it comprises:
Solid carrier, and
Be connected to the polypeptide according to claim 1 on this solid carrier.
3. detection means according to claim 2, wherein, described solid carrier is SJ modified silica-gel.
4. detection means according to claim 2, it is for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
5. a detection kit, it comprises the detection means according to any one of claim 2 ~ 4.
6. detection kit according to claim 5, it is for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
7. the purposes of polypeptide according to claim 1 in preparation detection kit.
8. purposes according to claim 7, wherein, described detection kit is used for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
9. purposes according to claim 7, wherein, described detection kit comprises the detection means according to any one of claim 2 ~ 4.
Apply the present invention to the clinical assistant diagnosis of enterovirns type 71 IgG antibody, gratifying effect can be obtained.
Accompanying drawing explanation
The schematic diagram that the making processes of Fig. 1 to SJ modified silica-gel (iPDMS film) is described.
Fig. 2 illustrates the schematic diagram of polypeptide microarrays spot sample mode.
Fig. 3 is the photo of display to the result that healthy normal people or non-bowel virus 71 type infected patient serum detect.
Fig. 4 is the photo of display to the result that enterovirns type 71 infected patient serum detects.
the embodiment of invention
Polypeptide of the present invention
In one aspect of the method, the invention provides the polypeptide useful to the clinical assistant diagnosis of enterovirns type 71 IgG antibody, be made up of the aminoacid sequence shown in SEQ ID NO:1, that is: QVVTVMDDLCQNPDGKDMSL.As shown in the Examples, the serum of polypeptide of the present invention to enterovirns type 71 infected patient is positive, and to be negative reaction to healthy normal people or non-bowel virus 71 type infected patient serum.Therefore, polypeptide of the present invention is useful as the clinical assistant diagnosis instrument of enterovirns type 71 IgG antibody.Polypeptide of the present invention may be used for manufacturing detection means (example detection means of the present invention described as follows) or the detection kit (example detection kit of the present invention described as follows) of the clinical assistant diagnosis that can be used for enterovirns type 71 IgG antibody.
Polypeptide of the present invention can be suitable for adopting the known method such as (1) chemical synthesis process or (2) enzyme reaction synthetic method to obtain, but is not limited thereto, and wherein chemosynthesis is more easy.In chemosynthesis polypeptide situation of the present invention, undertaken by using peptide synthesizer synthesis or this polypeptide semi-synthetic.As chemical synthesis process, such as peptide solid-phase synthesis etc. can be listed.The peptide of such synthesis can adopt conventional means such as ion-exchange chromatography, reverse phase high performance liquid chromatogram, affinity chromatography etc. to carry out purifying.Such peptide solid phase synthesis process with and subsequent peptide purification be all well-known in the art.
In addition, when producing polypeptide of the present invention by enzyme reaction, the method such as described in No. WO2004/011653, International Publication brochure can be adopted.Namely; can produce like this: by the amino acid of a side or the C-terminal of dipeptides is esterified or amidation and the amino acid that obtains or dipeptides, the amino acid (amino acid of such as carboxy protective) that is in unbound state with amino acid react under the existence of peptide synthetase, the dipeptides of generation or tripeptides.As peptide synthetase, can list: there is the culture of microorganism of the ability generating peptide, the bacterial disposing thing of the microbial cells be separated by this culture or this microorganism or this microbe-derived peptide synthetase.
And, except above-mentioned enzyme method, chemical synthesis process, such as gene engineering method can also be adopted to produce polypeptide of the present invention.
The aminoacid sequence of polypeptide of the present invention can adopt conventional method to carry out analyzing and confirming, such as, can utilize the method such as mass spectrum, chromatogram to carry out and analyze and confirm.
Detection means of the present invention
In another aspect, the present invention also provides a kind of detection means (detection means of the present invention), and it comprises solid carrier and the protein of the present invention that is connected on this solid carrier or polypeptide of the present invention.Described detection means is useful for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
In the present invention, solid carrier is not particularly limited, as long as the carrier of solid or insoluble material (be such as can pass through filtration, material that precipitation, magnetic resolution etc. are separated from reaction mixture).
The material forming solid carrier includes but not limited to: silica gel (polydimethylsiloxane, PDMS), Mierocrystalline cellulose, teflon
tM, Nitrocellulose, agarose, dextran, chitosan, polystyrene, polyacrylamide, polyester, polycarbonate, polymeric amide, polypropylene, nylon, poly(vinylidene fluoride), latex, silicon-dioxide, glass, glass fibre, gold, platinum, silver, copper, iron, stainless steel, ferrite, silicon wafer, polyethylene, polymine, poly(lactic acid), resin, polyose, albumen (albumin etc.), carbon or their combination etc.
The shape of solid carrier comprises but is not limited to: pearl, magnetic bead, film, microcapillary, filter membrane, plate, micro plate, carbon nanotube, sensor chip etc.Just as known in the art, the solid carrier that film or plate etc. are smooth can be arranged bottom pit, groove, filter membrane etc.
In the present invention, magnetic bead can have the sphere diameter of about 25nm ~ about 1mm scope.In a preferred embodiment, magnetic bead has the diameter of about 50nm ~ about 10 μm scope.The size of magnetic bead can be selected according to specific purposes.
In the present invention, the pearl be made up of the contour crosslinked spherical agarose of Sepharose has the diameter of about 24 μm ~ about 165 μm of scopes.Preferably, high crosslinked spherical sepharose 4B has the diameter of about 24 μm ~ about 44 μm of scopes.The size of high crosslinked spherical sepharose 4B can be selected according to specific purposes.
The example with the solid carrier of water repellent surface comprises can from Polysciences, Warrington, PA or Spherotech, the polystyrene latex beads such as the goods that Liberville, IL buy.
Silicon-dioxide (SiO
2)-process or silicon-dioxide (SiO
2) example of solid carrier of base comprises the extraordinary magnetic silica pearl etc. can bought from Polysciences, Warrington, PA, it may be used for catching nucleic acid (such as DNA).Or, the M-280 etc. that can buy from Dynal Biotech can also be used.
The magnetic bead with hydrophilic surface can be used for the bacterial cell of seizure proliferation period, nucleic acid and other composition.As the example of this magnetic bead, Polysciences can be listed, Warrington, the pearl (title: Biomag (registered trademark) carboxyl) that PA sells or Bangs Laboratory, Inc., the name of Fishers, IN is called the pearl of MC02N/2928.Or, the M-270 etc. that Dynal Biotech sells can be used.
In a preferred embodiment of the present invention, described solid carrier is SJ modified silica-gel, it is the microarray solid support material (iPDMS film, see the open CN101265329A of Chinese patent) of a kind of silicon rubber material of Suzhou Siju Biomaterials Co., Ltd.'s exploitation.This material is based on the conventional PDMS of biological study, add specific initiator composition (making this material realize surface-functionalized modification by surface initiated polymerization (SIP)) wherein, obtain through polyethylene glycol methacrylate-styrene polymer (poly (oligo (ethylene glycol) methacrylate), pOEGMA) finishing again.SJ modified silica-gel has outstanding anti-protein non-specific adsorption (Nonspecific protein adsorption, NPA) ability, non-specific protein absorption in complicated protein immunization can being detected controls to close to " absolute 0 " level (being near or below the limit of detection of instrument), not only can exempt the trouble closed and repeatedly clean, can also by the susceptibility using stronger amplification of signal means to improve protein microarray.And the essence of its silicon rubber imparts the stronger mechanical property of this material and good operability.Suzhou Yvonne gathers the combination assay microarray ELISA kit successfully SJ modified silica-gel being applied to 11 tumor markers compositions, achieve high-throughput and high-sensitive detection, demonstrating this material is a kind of outstanding protein microarray solid support material.Meanwhile, this material also has the adjustable characteristic of surface properties, can adjust its surface topography within the specific limits by the controlled modification reaction times.
The connection of polypeptide of the present invention and solid carrier can adopt the method for attachment that well known to a person skilled in the art polypeptide and solid carrier to carry out.Such as, for the connection on protein/polypeptide and modified silica-gel surface, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide [1-ethyl-3-(3-dimethyl ami-nopropyl) carbodiimide can be passed through, EDC] and N-hydroxy-succinamide (N-hydroxysuccinimide, NHS) reaction changes the carboxyl (-COOH) group on the macromolecular chain of modified silica-gel surface into activating group, this activating group can with protein/polypeptide on amino (-NH2) react thus realize protein/polypeptide being fixed on solid carrier surface (see such as Hongwei Ma, Yuanzi Wu, Xiaoli Yang, Xing Liu, Jian ' an He, Long Fu, Jie Wang, Hongke Xu, Yi Shi and Renqian Zhong,
integrated poly (dimethysiloxane) with an intrinsic nonfouling property approaching " Absolute " zero background in immunoassays, anal. Chem., 82,6338 – 6342,2010).
Concentration for polypeptide of the present invention in the sampling liquid used during point sample is not particularly limited, and those skilled in the art can conventionally select, and is preferably 1 μ g ~ 1000 μ g/mL, more preferably 10 μ g ~ 500 μ g/mL.In addition, the density distributed on a solid support for polypeptide of the present invention is not particularly limited, and those skilled in the art can conventionally select, and is preferably 1 ~ 100 points/10mm2, more preferably 5 ~ 50 points/10mm2.
Detection means of the present invention is useful for the clinical assistant diagnosis of enterovirns type 71 IgG antibody, or it may be used for manufacturing the detection kit of the clinical assistant diagnosis that can be used for enterovirns type 71 IgG antibody.
detection kit of the present invention
In one aspect of the method, the present invention also provides a kind of detection kit (detection kit of the present invention), and it comprises detection means of the present invention.This detection kit can be used for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
Detection kit of the present invention must comprise detection means of the present invention.Detection kit of the present invention can also comprise:
1. the serum sample diluent prepared or serum sample diluent component solution: Sample dilution, such as, have the Sample dilution (production code member 070021-S2) of Sai Chi bio tech ltd, Beijing, the application of sample variable color Sample dilution (production code member bwj010103) etc. of Bo Weijia bio tech ltd, Zhengzhou.Such Sample dilution is the PBS solution containing the composition such as BSA, sucrose, and containing sanitas, clarified liq, can directly use.This Sample dilution is used for dilute serum, and the serum that test kit detects will dilute suitable multiple, such as 2 ~ 200 times, preferably 10 ~ 100 times.
Detection kit of the present invention can also comprise:
2. concentrated washing lotion and washing lotion: after solid carrier surface hatches serum and ELIAS secondary antibody, the unconjugated antibody of solid carrier surface and ELIAS secondary antibody need be washed by washing lotion.Concentrated washing lotion is such as the polysorbas20 aqueous solution of 1%, need dilute 2 ~ 40 times, preferably 5 ~ 20 times during use.Can dilute by 1:9 by concentrating washing lotion (10 ×) purified water or distilled water, configure washing lotion.Such as, add 50 mL in 450 mL purified water or distilled water and concentrate washing lotion (10 ×), fully mix.The washing lotion do not used is placed on 2 ~ 8 DEG C of preservations, can preserve 3 months.
Detection kit of the present invention can also comprise:
3. ELIAS secondary antibody solution: the IgG in human enterovirus 71 infected person anteserum can be combined by the polypeptide of the present invention on solid carrier (such as SJ modified silica-gel), ELIAS secondary antibody can with antibodies, and the marker in ELIAS secondary antibody can react with luminous substrate mixing solutions, thus send detectable light.ELIAS secondary antibody can be the goat anti-human igg of such as horseradish peroxidase-labeled.Being not particularly limited the concentration of ELIAS secondary antibody in ELIAS secondary antibody solution, can be such as 1ng ~ 1000ng/mL.
Detection kit of the present invention can also comprise:
4. luminous substrate mixing solutions: luminous substrate mixing solutions can react with the horseradish peroxidase that marks in ELIAS secondary antibody, make to react the chemical light sending instrument and can detect, luminous substrate mixing solutions comprises luminous substrate liquid A-superoxol, and luminous substrate liquid B-luminol (luminol,3-aminophthalic acid cyclic hydrazide) solution.Adopt two kinds of luminous substrate liquid to mix with equal-volume during use in advance, obtain luminous substrate mixing solutions (using in 45 minutes).Luminol only has crosses just meeting luminescence by oxidizer treatment.The mixed aqueous solution of usual use hydrogen peroxide and a kind of hydroxide bases is as exciting agent.Under horseradish peroxidase enzyme catalytic, decomposing hydrogen dioxide solution is oxygen G&W:
2 H
2O
2→ O
2+ 2 H
2O
Luminol,3-aminophthalic acid cyclic hydrazide and oxyhydroxide generate a pairs of anion when reacting, the dioxygen oxidation that it can be gone out by peroxide decomposition, and product is an organo-peroxide.This superoxide is very unstable, decomposites nitrogen immediately, generates the 3-aminophthalic acid of excited state.During excited state to ground state transforms, the energy of release exists with the form of photon, and wavelength is positioned at the blue light components of visible ray.The SuperSignal ELISA Femto Maximum Sensitivi-of the example such as Thermo Seientific company of luminous substrate mixing solutions
Ty Substrate, article No. 37074.
Detection kit of the present invention can also comprise:
5. one or more reaction cavity, can adopt such as two-sided biological incubation reactor, Chinese patent ZL 201120177686.3 or ZL201110142518.5 according to demand; Or the biological incubation reactor ZL201220430886.x of one side.
Detection kit of the present invention can also comprise:
6. other are for detecting the detection molecules (such as polypeptide, protein, nucleic acid etc.) of enterovirns type 71.
Detection kit of the present invention can also comprise:
7. working instructions, wherein describe the clinical assistant diagnosis that this detection kit may be used for enterovirns type 71 IgG antibody.
Reaction member and reaction kit can such as adopt gel imaging instrument to carry out chemiluminescence imaging.As gel imaging instrument, such as GE LAS4000mini type or sky energy 5500 type Microarray image instrument can be adopted.
Embodiment
Below, by embodiment, more specific description is carried out to the present invention, but be not the restriction to the technology of the present invention scope.By the record of this specification sheets, those skilled in the art can be easy to modify the present invention/change, and these are included in technical scope of the present invention.Scope of the present invention is determined by claims.
1. polypeptide preparation and confirmation
The polypeptide used in embodiment has the aminoacid sequence shown in SEQ ID NO:1, and by Shanghai gill, biochemical company limited synthesizes, and the sign of this polypeptide confirms to have synthesized described polypeptide by hydrogen spectrum and mass spectrum.Liquid phase chromatography detects purity: 95.31% (area normalization method): mass spectroscopy detects (ESI-MS): calculate molecular weight: 2208.52, test value: 2208.06.
2. the preparation of detection means
Detection chip be with SJ modified silica-gel (iPDMS film) for solid support material, be prepared from by point sample immobilized polypeptide solution thereon.Modified silica-gel be add in traditional polydimethyl siloxane material band olefin-terminal, the initiator of surface initiated polymerization, and be fixed in the three-dimensional structure of polydimethylsiloxane by heat cross-linking (si-h bond bonding), obtain SJ modified silica-gel, its making processes as shown in Figure 1.
A and B is wherein two components of polydimethylsiloxane, polydimethylsiloxane (Poly (dimethylsiloxane), Sylgard 184) buy from Dow corning (DowCorning) company, comprise liquid composition A (composition is metallic platinum catalyst and the diformazan siloxanes polymer precursor mixture being with vinyl) and crosslinking agent B (composition is the dimethyl siloxane precursor with vinyl and Si-H group) two kinds of compositions.C is the initiator of end strips vinyl, is purchased from Hangzhou Dong Wei company.Polymer on finally modifying is that oligomeric ethylene glycol methacrylate monomer (Oligo (ethylene glycol) methacrylate, hereinafter referred to as OEGMA, molecular weight Mw=526) is bought in Aldrich.Polydimethylsiloxane precursor A and crosslinking agent B are fully mixed with A:B:C=10:1:0.5 ratio with the initiator C of band vinyl end.Make transparent elastic silicone rubber by curing reaction, then carry out finishing by SIP technology and can obtain SJ modified silica-gel.Experiment shows, the surface of SJ modified silica-gel have enough highdensity, by the initiator of covalently immobolization, it can pass through surface initiated polymerization (SIP), and to realize surface macromolecule modified.The surface that reaction acquisition polyoxyethylene glycol (Polyethylene Glycol, PEG) is modified is carried out in use poly (OEGMA) (polyethylene glycol methacrylate-styrene polymer), realizes the ability of stronger anti-albumen non-specific adsorption.
The SJ modified silica-gel film made need be kept in 4 DEG C of refrigerators.
Adopt brilliant core PersonalArrayerTM 16 people's point sample instruments to prepare polypeptide microarrays on modified silica-gel, process is:
1) pre-treatment
By SJ modified silica-gel thin slice (15 × 15mm
2) be immersed in activation solution, take out with deionized water drip washing 3 times after 30min, dry up with nitrogen, be used for point sample at once.
2) point sample
Sampling liquid is diluted and gets well and transfer in the corresponding micropore of 384 orifice plate, 384 orifice plates of band sample are placed on point sample instrument base station, pretreated modified silica-gel thin slice are placed on the base station of point sample instrument simultaneously, carry out point sample at once.Point sample envrionment conditions is room temperature (25 DEG C), and humidity set is 50%.On the polypeptide microarrays made, the point sample amount of each point is about 0.6nL, and sampling point radius is 200 μm.
3) chemistry is fixing
The polypeptide microarrays just made will be placed on fixing at least 6h in climatic chamber (26 DEG C, 60% humidity).Chemistry fixation procedure is as follows:
First by point sample instrument by include catch peptide molecule damping fluid point on modified silica-gel film, then damping fluid starts evaporation, catch peptide molecule and the surface intimate contact interacting of SJ modified silica-gel, by Chemical bond, the high molecular end-COOH of ploy (OEGMA) on modified silica-gel surface and peptide molecule--NH
2formed and stablize covalent linkage, and then chemically active peptide molecule will be had to be fixed on SJ modified silica-gel surface.
4) assemble
The polypeptide microarrays of fixing 6h must assemble in two days.First by gum, SJ modified silica-gel thin slice is attached on special reaction column, covers reaction cavity.A reactor is made up of two reaction columns and a reaction cavity.
5) preserve
The polypeptide microarrays assembled, needs to vacuumize sealing, is kept in the refrigerator of 4 DEG C, for subsequent use.
3. detect by detection means
Checking procedure
1, before starting to detect, concentrated washing lotion is added purified water in the ratio of 1:10 or distilled water dilutes, must washing lotion after dilute, direct use, use liquid-transfering gun that 2mL washing lotion is added to chip surface, soak chip 3 minutes, ensure that chip surface is fully wet out.
2, test serum sample Sample dilution is mixed according to 1:40 dilution.
3, discard the washing lotion of soaking chip, under the state that chip surface is completely moistening, the serum after each serum sample draws 200 μ L dilutions joins in chip reactor.
4, chip reactor is put into chip permanent seat, be put on shaking table, open shaking table, frequency 150 revs/min, incubated at room 30 minutes.
5, the serum sample in chip reactor is discarded, with 15mL washing lotion cleaning reaction cavity and chip surface 3 times.
6, after having cleaned, each chip reactor adds 200 μ L ELIAS secondary antibody solution respectively, chip reactor is put into chip permanent seat, is put on shaking table, opens shaking table, frequency 150 revs/min, incubated at room 30 minutes.
7, the ELIAS secondary antibody solution in chip reactor is discarded, with 15mL washing lotion cleaning reaction cavity and chip surface 3 times.
8, in advance luminous substrate liquid A and luminous substrate liquid B can be mixed with equal-volume, obtain luminous substrate mixing solutions (using in 45 minutes).To be cleaned complete after, take off reaction cavity, each chip surface adds 15 μ L luminous substrate mixing solutionss respectively, makes luminous substrate mixing solutions can be laid on chip surface uniformly.
9, the chip adding luminescent solution is placed in gel imaging instrument chemiluminescence imaging, and sentence read result.
Healthy normal human serum, enterovirns type 71 infected patient serum and other disease patient's serum samples are provided by chain hospital, and disease criterion all through Clinical Laboratory confirmation, all obtains the Informed Consent Form of supplier.
Negative control has PBS damping fluid (namely to hatch without test serum in the 3rd step, and hatch by PBS solution, all the other steps are identical) contrast, the contrast of serum dilution, and negative serum (refer to Healthy People and non-bowel virus 71 type infected patients serum) contrast.
The spot sample mode of polypeptide microarrays is as shown in Figure 2: wherein, the sample of 16 points of black circle is human IgG, as locating point; The sample of foursquare 4 points is PB sampling liquid, as the blank of experiment; The sample of white circular form point is other enterovirns type 71 autoantigen protein polypeptide, as Testing index (these polypeptide have response to illustrate in detected serum have enterovirns type 71 autoantibody); The sample polypeptides of star point is polypeptide of the present invention, and it is enterovirns type 71 autoantigen protein polypeptide, can produce response to enterovirns type 71 infected patient serum.
This polypeptide microarrays is used to detect enterovirns type 71 infected patient serum and negative control according to above-mentioned detecting step, response modes is as shown in Fig. 3 ~ 4: wherein, Fig. 3 shows the detected result of negative control, only has the sample of the point shown in black circle to have response.Fig. 4 shows the detected result of enterovirns type 71 infected patient serum, and black is circular, white is circular and sample that is star point has response.It should be noted that, instrument records signal value from low to high, and corresponding signaling point color is by black-red-white gradual change.
Sequence table
<110> Zhao Ya duckweed
<120> polypeptide, comprise the detection means of this polypeptide and comprise the detection kit of this device
<130> E04-044
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 20
<212> PRT
<213> artificial sequence
<220>
<221> misc_feature
<223> Enterovirus 71 antigen polypeptide
<400> 1
Gln Val Val Thr Val Met Asp Asp Leu Cys Gln Asn Pro Asp Gly Lys
1 5 10 15
Asp Met Ser Leu
20
Claims (9)
1. a peptide species, its aminoacid sequence as shown in SEQ ID NO:1, that is: QVVTVMDDLCQNPDGKDMSL.
2. a detection means, it comprises:
Solid carrier, and
Be connected to the polypeptide according to claim 1 on this solid carrier.
3. detection means according to claim 2, wherein, described solid carrier is SJ modified silica-gel.
4. detection means according to claim 2, it is for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
5. a detection kit, it comprises the detection means according to any one of claim 2 ~ 4.
6. detection kit according to claim 5, it is for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
7. the purposes of polypeptide according to claim 1 in preparation detection kit.
8. purposes according to claim 7, wherein, described detection kit is used for the clinical assistant diagnosis of enterovirns type 71 IgG antibody.
9. purposes according to claim 7, wherein, described detection kit comprises the detection means according to any one of claim 2 ~ 4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410848156.5A CN104447959A (en) | 2014-12-31 | 2014-12-31 | Polypeptide, detection device containing polypeptide and detection reagent kit provided with device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410848156.5A CN104447959A (en) | 2014-12-31 | 2014-12-31 | Polypeptide, detection device containing polypeptide and detection reagent kit provided with device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104447959A true CN104447959A (en) | 2015-03-25 |
Family
ID=52894766
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410848156.5A Pending CN104447959A (en) | 2014-12-31 | 2014-12-31 | Polypeptide, detection device containing polypeptide and detection reagent kit provided with device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104447959A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004104189A1 (en) * | 2003-05-21 | 2004-12-02 | Amsterdam Institute Of Viral Genomics B.V. | New enterovirus, vaccines, medicaments and diagnostic kits |
| CN102539776A (en) * | 2010-12-10 | 2012-07-04 | 浙江普康生物技术股份有限公司 | Enzyme-linked immunosorbent assay method for enterovirus 71-specific antibody |
| CN102558313A (en) * | 2012-01-12 | 2012-07-11 | 东南大学 | Enterovirus 71 type specific recombinant protein antigen and application thereof |
-
2014
- 2014-12-31 CN CN201410848156.5A patent/CN104447959A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004104189A1 (en) * | 2003-05-21 | 2004-12-02 | Amsterdam Institute Of Viral Genomics B.V. | New enterovirus, vaccines, medicaments and diagnostic kits |
| CN102539776A (en) * | 2010-12-10 | 2012-07-04 | 浙江普康生物技术股份有限公司 | Enzyme-linked immunosorbent assay method for enterovirus 71-specific antibody |
| CN102558313A (en) * | 2012-01-12 | 2012-07-11 | 东南大学 | Enterovirus 71 type specific recombinant protein antigen and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| KOIKE,S.: "accession BAG82822.1", 《GENBANK》 * |
| 李玉娇等: "抗原优势表位的筛选方法", 《中国病原生物学杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105732772A (en) | Polypeptide, detection device containing polypeptide and detection kit containing device | |
| CN105732775A (en) | Polypeptide, detection device containing polypeptide and detection kit containing device | |
| CN104945481A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN105732771A (en) | Polypeptide, detection device containing polypeptide and detection kit containing device | |
| CN105732776A (en) | Polypeptide, detection device containing polypeptide and detection kit containing device | |
| CN105367626A (en) | Polypeptide, detection device and detection kit for detecting cervical cancer | |
| CN104945472A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN104945474A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN104945476A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN104945477A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN105732774A (en) | Polypeptide, detection device containing polypeptide and detection kit containing device | |
| CN105732769A (en) | Polypeptides, detection device comprising the polypeptides, and detection kit comprising the detection device | |
| CN105732773A (en) | Polypeptide, detection device containing polypeptide and detection kit containing device | |
| CN105732770A (en) | Polypeptides, a detection device comprising the polypeptides, and a detection kit comprising the detection device | |
| CN104447959A (en) | Polypeptide, detection device containing polypeptide and detection reagent kit provided with device | |
| CN104945480A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN104945473A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN104945475A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN103880923A (en) | Polypeptide, detector containing the polypeptide, and detection kit containing the polypeptide | |
| CN104725479A (en) | Polypeptide, detection device comprising polypeptide, and detection kit | |
| CN103880922B (en) | Polypeptide, the detection means comprising this polypeptide and detection kit | |
| CN104945482A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN103965313B (en) | Polypeptide, the detection means comprising this polypeptide and detection kit | |
| CN104945478A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device | |
| CN104945479A (en) | Polypeptide, detection device comprising polypeptide, and detection kit comprising device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150325 |
|
| WD01 | Invention patent application deemed withdrawn after publication |