CN1778949A - Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase - Google Patents
Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase Download PDFInfo
- Publication number
- CN1778949A CN1778949A CN 200410065564 CN200410065564A CN1778949A CN 1778949 A CN1778949 A CN 1778949A CN 200410065564 CN200410065564 CN 200410065564 CN 200410065564 A CN200410065564 A CN 200410065564A CN 1778949 A CN1778949 A CN 1778949A
- Authority
- CN
- China
- Prior art keywords
- phosphonuclease
- inosine
- reagent
- ribose
- nucleotidase
- 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
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 58
- 230000000694 effects Effects 0.000 title claims description 21
- 102000004008 5'-Nucleotidase Human genes 0.000 title claims description 15
- 108010043671 prostatic acid phosphatase Proteins 0.000 title claims description 15
- 239000005515 coenzyme Substances 0.000 claims abstract description 34
- 238000000034 method Methods 0.000 claims abstract description 28
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 52
- 229930010555 Inosine Natural products 0.000 claims description 43
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 claims description 43
- 229960003786 inosine Drugs 0.000 claims description 43
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 claims description 33
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 claims description 33
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 claims description 33
- 238000006243 chemical reaction Methods 0.000 claims description 28
- 238000013016 damping Methods 0.000 claims description 27
- 239000012530 fluid Substances 0.000 claims description 27
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 26
- 108010058987 inosine nucleosidase Proteins 0.000 claims description 17
- 239000000872 buffer Substances 0.000 claims description 12
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 238000009007 Diagnostic Kit Methods 0.000 claims description 11
- 239000002253 acid Substances 0.000 claims description 9
- 230000001186 cumulative effect Effects 0.000 claims description 9
- 230000035484 reaction time Effects 0.000 claims description 9
- RDNLUQQRUBTKQB-VEGRVEBRSA-N (2r,3r,4r)-2,3,4,5-tetrahydroxypentanal;hydrate Chemical compound O.OC[C@@H](O)[C@@H](O)[C@@H](O)C=O RDNLUQQRUBTKQB-VEGRVEBRSA-N 0.000 claims description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 4
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 claims description 4
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 3
- 229960004418 trolamine Drugs 0.000 claims description 3
- CXONXVMMINSQBV-NNYOXOHSSA-N (2r,3r,4s,5r)-5-[[[[(2r,3s,4r,5r)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl]oxymethyl]-2-(3-carbamothioylpyridin-1-ium-1-yl)-4-hydroxyoxolan-3-olate Chemical compound NC(=S)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)[O-])=C1 CXONXVMMINSQBV-NNYOXOHSSA-N 0.000 claims description 2
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical group NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 claims description 2
- 150000004676 glycans Chemical class 0.000 claims description 2
- 235000011187 glycerol Nutrition 0.000 claims description 2
- 229960003966 nicotinamide Drugs 0.000 claims description 2
- 235000005152 nicotinamide Nutrition 0.000 claims description 2
- 239000011570 nicotinamide Substances 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 150000005846 sugar alcohols Polymers 0.000 claims description 2
- YMAWOPBAYDPSLA-UHFFFAOYSA-N glycylglycine Chemical compound [NH3+]CC(=O)NCC([O-])=O YMAWOPBAYDPSLA-UHFFFAOYSA-N 0.000 claims 2
- 229940058020 2-amino-2-methyl-1-propanol Drugs 0.000 claims 1
- PDLNHDSYGLTYDS-UHFFFAOYSA-N 3-aminopropanoic acid;hydrochloride Chemical compound Cl.NCCC(O)=O PDLNHDSYGLTYDS-UHFFFAOYSA-N 0.000 claims 1
- 108010008488 Glycylglycine Proteins 0.000 claims 1
- CBTVGIZVANVGBH-UHFFFAOYSA-N aminomethyl propanol Chemical compound CC(C)(N)CO CBTVGIZVANVGBH-UHFFFAOYSA-N 0.000 claims 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims 1
- 235000011130 ammonium sulphate Nutrition 0.000 claims 1
- 229940043257 glycylglycine Drugs 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 14
- 239000007853 buffer solution Substances 0.000 abstract description 6
- 238000005859 coupling reaction Methods 0.000 abstract description 5
- 102000004190 Enzymes Human genes 0.000 abstract description 3
- 108090000790 Enzymes Proteins 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000010339 medical test Methods 0.000 abstract description 2
- 238000002156 mixing Methods 0.000 abstract description 2
- 238000002835 absorbance Methods 0.000 abstract 1
- 230000004913 activation Effects 0.000 abstract 1
- 150000004712 monophosphates Chemical class 0.000 abstract 1
- 239000002773 nucleotide Substances 0.000 abstract 1
- 125000003729 nucleotide group Chemical group 0.000 abstract 1
- 239000000376 reactant Substances 0.000 abstract 1
- 239000003381 stabilizer Substances 0.000 abstract 1
- 239000000203 mixture Substances 0.000 description 9
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 230000008878 coupling Effects 0.000 description 4
- 238000010168 coupling process Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000011574 phosphorus Substances 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 3
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 3
- 238000001976 enzyme digestion Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 description 2
- FTOAOBMCPZCFFF-UHFFFAOYSA-N 5,5-diethylbarbituric acid Chemical compound CCC1(CC)C(=O)NC(=O)NC1=O FTOAOBMCPZCFFF-UHFFFAOYSA-N 0.000 description 2
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 description 2
- 206010008635 Cholestasis Diseases 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 231100000359 cholestasis Toxicity 0.000 description 2
- 230000007870 cholestasis Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 208000006454 hepatitis Diseases 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 150000002460 imidazoles Chemical class 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- VXEQGQXRKQSAMW-UHFFFAOYSA-N 2-amino-2-methylpropan-1-ol Chemical compound CC(C)(N)CO.CC(C)(N)CO VXEQGQXRKQSAMW-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 206010008909 Chronic Hepatitis Diseases 0.000 description 1
- UDMBCSSLTHHNCD-UHFFFAOYSA-N Coenzym Q(11) Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(O)=O)C(O)C1O UDMBCSSLTHHNCD-UHFFFAOYSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- YTNIXZGTHTVJBW-SCRDCRAPSA-N FMNH2 Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2NC2=C1NC(=O)NC2=O YTNIXZGTHTVJBW-SCRDCRAPSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 208000024815 Granulomatous liver disease Diseases 0.000 description 1
- 206010019695 Hepatic neoplasm Diseases 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 206010023129 Jaundice cholestatic Diseases 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- ZQYUHRVUJCLGRX-UHFFFAOYSA-N NCC(=O)NCC(O)=O.NCC(=O)NCC(O)=O Chemical compound NCC(=O)NCC(O)=O.NCC(=O)NCC(O)=O ZQYUHRVUJCLGRX-UHFFFAOYSA-N 0.000 description 1
- 201000005267 Obstructive Jaundice Diseases 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 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
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 1
- UDMBCSSLTHHNCD-KQYNXXCUSA-N adenosine 5'-monophosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H]1O UDMBCSSLTHHNCD-KQYNXXCUSA-N 0.000 description 1
- 229950006790 adenosine phosphate Drugs 0.000 description 1
- 229960002319 barbital Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 1
- GHXRKGHKMRZBJH-UHFFFAOYSA-N boric acid Chemical compound OB(O)O.OB(O)O GHXRKGHKMRZBJH-UHFFFAOYSA-N 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- UETQVDZZPKAQIC-UHFFFAOYSA-N chlorane Chemical compound Cl.Cl.Cl.Cl UETQVDZZPKAQIC-UHFFFAOYSA-N 0.000 description 1
- 208000003167 cholangitis Diseases 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 229940111685 dibasic potassium phosphate Drugs 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- VWWQXMAJTJZDQX-UYBVJOGSSA-N flavin adenine dinucleotide Chemical compound C1=NC2=C(N)N=CN=C2N1[C@@H]([C@H](O)[C@@H]1O)O[C@@H]1CO[P@](O)(=O)O[P@@](O)(=O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C2=NC(=O)NC(=O)C2=NC2=C1C=C(C)C(C)=C2 VWWQXMAJTJZDQX-UYBVJOGSSA-N 0.000 description 1
- 235000019162 flavin adenine dinucleotide Nutrition 0.000 description 1
- 239000011714 flavin adenine dinucleotide Substances 0.000 description 1
- 229940093632 flavin-adenine dinucleotide Drugs 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229960001031 glucose Drugs 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-M glutaminate Chemical compound [O-]C(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-M 0.000 description 1
- 208000017694 hepatic granuloma Diseases 0.000 description 1
- 231100000843 hepatic granuloma Toxicity 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 230000000155 isotopic effect Effects 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- QVLTXCYWHPZMCA-UHFFFAOYSA-N po4-po4 Chemical compound OP(O)(O)=O.OP(O)(O)=O QVLTXCYWHPZMCA-UHFFFAOYSA-N 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229950001574 riboflavin phosphate Drugs 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Substances [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- WUWHFEHKUQVYLF-UHFFFAOYSA-M sodium;2-aminoacetate Chemical compound [Na+].NCC([O-])=O WUWHFEHKUQVYLF-UHFFFAOYSA-M 0.000 description 1
- OTNVGWMVOULBFZ-UHFFFAOYSA-N sodium;hydrochloride Chemical compound [Na].Cl OTNVGWMVOULBFZ-UHFFFAOYSA-N 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- -1 sulfuric acid amine Chemical class 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 description 1
- YNIRKEZIDLCCMC-UHFFFAOYSA-K trisodium;phosphate;hydrate Chemical compound [OH-].[Na+].[Na+].[Na+].OP([O-])([O-])=O YNIRKEZIDLCCMC-UHFFFAOYSA-K 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention is about a method of measuring the activation of 5'-phosphonuclease, and it also concerns the reagent box of 5'-phosphonucleasediagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, carnine monophosphate, oxidized coenzyme, carnine nucleotide, ribose1-dehyddrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get theactivation of 5'-phosphonuclease. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.
Description
Technical field
The present invention relates to a kind ofly to measure 5 '-method of activity of 5 '-nucleotidase, the invention still further relates to simultaneously be used to realize 5 of this method '-the phosphonuclease diagnostic kit, belong to medical test determination techniques field.
Background technology
Medical research shows, 5 '-phosphonuclease (5NT) increases and is mainly seen in the obstructive jaundice, also is found in liver cancer and hepatitis.When the concurrent cholangitis of cholestasis, primary and Secondary cases cholehepatocirrhosis and chronic hepatitis, the 5NT rate of rise is higher than alkaline phosphatase; The susceptibility that 5NT raises when liver tumor and hepatic granuloma is higher than alkaline phosphatase.Because 5 '-activity of 5 '-nucleotidase do not have physiological and raise, and is not only responsive than alkaline phosphatase for diagnosis infant's hepatopathy and gestational liver function cholestasis, and specificity is arranged.Therefore measure 5 '-activity of phosphonuclease is significant for the diagnosis of disease.
5 '-activity determination method of phosphonuclease is a lot, mainly contains isotope substrate method, chemical phosphorus acid test method (nineteen twenty-five).Understand according to the applicant, generally adopt the isotropic substance Substrate test method at present in the world, method is: 5 '-phosphonuclease acts on H
3-dUMP, after the termination reaction,, try to achieve 5 by the ion exchange column analytical results '-activity of 5 '-nucleotidase.Perhaps utilize 5 '-phosphonuclease produces inorganic phosphorus after acting on single adenosine phosphate, analyze content of inorganic phosphorus with Chemical acid method again and calculated 5 '-activity of phosphonuclease.
Method of isotope substrate is complicated to also have isotopic contamination, and needs Isotope analyzer, therefore is difficult to apply conscientiously.Determination of inorganic phosphorus is the method for nineteen twenty-five invention, and accuracy is bad, and strong acid contaminate environment or the like also is not suitable for applying.
Summary of the invention
Propose a kind of can overcome 5 of above prior art shortcoming '-measuring method of activity of 5 '-nucleotidase, provide simultaneously in order to realize 5 of this method '-the phosphonuclease diagnostic kit.Adopt reagent in this test kit not only can on ultraviolet analyser or half, automatic clinical chemistry analyzer, carry out 5 '-activity of 5 '-nucleotidase is measured, and finding speed is fast, accuracy is high, thereby can obtain practical applying.
The present invention measures 5 '-step of activity of 5 '-nucleotidase is as follows:
1), with sample and the reagent mix of mainly forming by inosine list phosphoric acid, oxidized coenzyme, Inosine nucleosidase, ribose 1-desaturase, make it to take place the reaction of following principle:
Inosine list phosphoric acid+water
5 '-phosphonucleaseInosine+single phosphoric acid
Inosine+water
Inosine nucleosidaseRibose+uridylic
Ribose+oxidized coenzyme+water
Ribose 1-desaturaseRibonic acid+reduced coenzyme
2), detect the end reaction thing in the speed that predominant wavelength 340nm absorbancy rises, calculate 5 '-the active size of phosphonuclease.
Usually step 2) the end reaction thing is placed under ultraviolet analyser or half, the automatic clinical chemistry analyzer device, detect the speed that predominant wavelength 340nm absorbancy rises, calculate 5 '-the active size of phosphonuclease.
The blending ratio of above sample and reagent by volume is 1/10 to 1/250, and the mensuration temperature of above process is controlled at 20 ℃ to 50 ℃ conventional scopes, and the reaction times was controlled at conventional 2-30 minute.
The present invention uses 5 '-phosphonuclease coupling Inosine nucleosidase (EC 3.2.2.2), ribose 1-desaturase (EC 1.1.1.115) reaction continuous monitoring method.5 '-phosphonuclease hydrolysis inosine list phosphoric acid generation inosine, Inosine nucleosidase catalysis inosine produces ribose then, ribose is the effect by coupling ribose 1-desaturase again, with oxidized coenzyme (not having absorption peak) reduction becoming dihydrocoenzyme (absorption peak being arranged) at the 340nm place at the 340nm place, thereby measured the speed that reduced coenzyme rises in 340nm place absorbancy, by measuring the speed that 340nm place absorbancy rises, can calculate 5 '-phosphonuclease active big or small.
5 of realization the inventive method '-the phosphonuclease diagnostic kit can be single agent, comprising:
Damping fluid 40--200mmol/l
Inosine list phosphoric acid 0.5--50mmol/l
Oxidized coenzyme 0.5--10mmo/l
Inosine nucleosidase 500--50000U/l
Ribose 1-desaturase 500--50000U/l
Stablizer 10--80% (cumulative volume)
Also above single agent following pair of agent be can be made into, inside and outside source inosine, ribose pollution more helped eliminating:
Reagent I
Damping fluid 40--200mmol/l
Oxidized coenzyme 0.5--10mmo/l
Inosine nucleosidase 500--50000U/l
Ribose 1-desaturase 500--50000U/l
Stablizer 10--80% (cumulative volume)
Reagent II
Damping fluid 40--200mmol/l
Inosine list phosphoric acid 0.5--50mmol/l
Stablizer 10--50mmol/l
The prescription of two agent is not limited only to above-mentioned prescription, the composition of reagent I wherein, and oxidized coenzyme, Inosine nucleosidase, ribose 1-desaturase etc. can be placed on reagent II, reagent II composition wherein, inosine list phosphoric acid also can be placed on reagent I, so can form multiple formulations, does not describe in detail one by one at this.
Reagent can also be made into following three reagent, not only more help eliminating inside and outside source inosine, ribose pollution, also more help the stable of reagent:
Reagent I
Damping fluid 40--200mmol/l
Oxidized coenzyme 0.5--10mmo/l
Stablizer 10--50mmol/l
Reagent II
Damping fluid 40--200mmol/l
Inosine nucleosidase 500--50000U/l
Ribose 1-desaturase 500--50000U/l
Stablizer 10--80% (cumulative volume)
Reagent III
Damping fluid 40--200mmol/l
Inosine list phosphoric acid 0.5--50mmol/l
Stablizer 10--50mmol/l
Three doses prescription is not limited only to above-mentioned prescription, and the composition of reagent I wherein, oxidized coenzyme can be placed among reagent II or the reagent III.Reagent II composition wherein, Inosine nucleosidase, ribose 1-desaturase etc. also can be placed among reagent I or the reagent III.Reagent III composition wherein, inosine list phosphoric acid also can be placed among reagent I or the reagent II, so can form multiple formulations, does not describe in detail one by one.
The pH scope of buffer reagent can be 6.0～11.0.Buffer reagent can be three (carboxymethyl) aminomethane one hydrochloric acid (Tris-HCl) damping fluid, phosphoric acid salt (Phosphate) damping fluid, trolamine (Triethanolamine) damping fluid, 2-amino-2-methyl-1-propanol (2-Amino-2-methyl-1-propanol) damping fluid, imidazoles (Imidazole) damping fluid or glycylglycine (Glycylglycine) damping fluid etc., (for example can also be imidazoles/hydrochloride buffer 6.2-7.8 but be not limited only to these damping fluids; Sodium phosphate dibasic citrate buffer solution 5.0-8.0; Citric acid sodium citrate buffer solution 5.0-6.6; Phosphate buffered saline buffer 6.0-8.0; Sodium phosphate dibasic-potassium phosphate buffer 6.0-8.0; Potassium primary phosphate sodium hydrate buffer solution 6.0-8.0; Veronal sodium-hydrochloride buffer 6.8-9.6; Tris hydrochloride buffer 7.0-9.0; Boric acid borate buffer solution 7.4-9.0; Glycine-sodium hydrate buffer solution 8.6-10.6; Sand-sodium hydrate buffer solution 9.2-10.0; Yellow soda ash-sodium bicarbonate buffer liquid 8.8-10.6 (Ca
2+, Mg
2+Must not use when existing); PBS damping fluid 7.0-7.6 etc.).
Above oxidized coenzyme can be NADP
+, NAD
+Or thio-NAD
+Deng oxidized form nicotinamide coenzyme or derivatives thereof.
In addition, in order to reduce the cross influence between each reagent composition, the stability that keeps reagent, so that standing storage, above single agent, the reagent I of two agent, the reagent I of reagent II or three doses, reagent II, usually add stablizer 10～80% or 10～50mmol/l among the reagent III, stablizer can be an ethylene glycol, propylene glycol, glycerine, glycan, polyalcohols, in sulfuric acid amine or the salt etc. one or more can also be at thioglycol, adenosine diphosphate (ADP), bovine serum albumin, carbonate, cholate, dextran, ethylenediamine tetraacetic acid (EDTA), flavin adenine dinucleotide, vitamin B2 phosphate, glucose, glutaminate, lactose, N.F,USP MANNITOL, sucrose, sodium-chlor, select in the succinate etc.
Experiment shows, takes all factors into consideration from the accuracy of measurement result and economy two aspects of preparation cost, no matter is single agent, two agent or three doses, the present invention 5 of following system component relation '-the phosphonuclease diagnostic kit is comparatively desirable:
Damping fluid 80--120mmol/l
Inosine list phosphoric acid 2--10mmol/l
Oxidized coenzyme 2--6mmo/l
Inosine nucleosidase 5000--10000U/1
Ribose 1-desaturase 5000--10000U/l
Stablizer 20--50% (cumulative volume)
Because the present invention utilizes Enzymology method, enzyme digestion reaction to have the high characteristics of specificity fully, is not vulnerable to the interference of other materials of inside and outside source.Easy, the easy handling of enzyme process method.The specificity of enzyme digestion reaction impels test result accurate.Enzyme digestion reaction all is to carry out under buffer conditions, does not have environmental pollution problem.The enzyme process method does not need special, extra instrument, and testing cost is cheap.Therefore can guarantee to have higher test accuracy, be more convenient for applying.
In addition, one of striking features of the present invention is the pollution that can eliminate inside and outside source inosine, ribose, the effect of eliminating inside and outside source inosine, ribose occurs in the first half of entire reaction time period, be consumed totally at time second half section contaminated inside and outside source inosine, ribose, and second half section time test 5 '-the needed inosine of activity of 5 '-nucleotidase, ribose all be result from 5 '-activity of phosphonuclease.
Embodiment
The present invention is further illustrated below in conjunction with examples of implementation.
Embodiment one (single agent)
5 of present embodiment '-the phosphonuclease diagnostic kit comprises:
Damping fluid 80mmol/l
Inosine list phosphoric acid 2mmol/l
Oxidized coenzyme 2mmo/l
Inosine nucleosidase 5000U/l
Ribose 1-desaturase 5000U/l
Stablizer 50% (cumulative volume)
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction times, test predominant wavelength 340nm, more than the test commplementary wave length 405nm, tested 5 '-volume ratio of phosphonuclease sample and reagent is 1/25, the Direction of Reaction is positive reaction, 1 minute time of lag, 2 minutes detection times, theoretical k value-4180.
After adding sample and reagent, make it to mix, following reaction take place:
Inosine list phosphoric acid+water
5 '-phosphonucleaseInosine+single phosphoric acid
Inosine+water
Inosine nucleosidaseRibose+uridylic
Ribose+oxidized coenzyme+water
Ribose 1-desaturaseRibonic acid+reduced coenzyme
The end reaction thing is placed under the biochemical analyzer, detect the speed that predominant wavelength 340nm absorbancy rises, thereby calculate 5 '-the active size of phosphonuclease.
Present embodiment uses 5 '-phosphonuclease coupling Inosine nucleosidase (EC 3.2.2.2), ribose 1-desaturase (EC 1.1.1.115) reaction continuous monitoring method.5 '-phosphonuclease hydrolysis inosine list phosphoric acid generation inosine, Inosine nucleosidase catalysis inosine produces ribose then, ribose is the effect by coupling ribose 1-desaturase again, with oxidized coenzyme (not having absorption peak) reduction becoming dihydrocoenzyme (absorption peak being arranged) at the 340nm place at the 340nm place, thereby measured the speed that reduced coenzyme rises in 340nm place absorbancy, by measuring the speed that 340nm place absorbancy rises, can calculate 5 '-phosphonuclease active big or small.
Embodiment two (two agent)
5 of present embodiment '-the phosphonuclease diagnostic reagent has:
Reagent I
Damping fluid 100mmol/l
Oxidized coenzyme 4mmo/l
Inosine nucleosidase 8000U/l
Ribose 1-dehydrogenase 8 000U/l
Stablizer 50% (cumulative volume)
Reagent II
Damping fluid 100mmol/l
Inosine list phosphoric acid 6mmol/l
Stablizer 20mmol/l
Measure 5 '-during activity of 5 '-nucleotidase, temperature is controlled at 30 ℃, 15 minutes reaction times, test predominant wavelength 340nm, more than the test commplementary wave length 405nm, tested 5 '-volume ratio of phosphonuclease sample and reagent is 1/25, the Direction of Reaction is positive reaction, 1 minute time of lag, 2 minutes detection times, theoretical k value-4180.
Concrete determination step is:
Inosine list phosphoric acid+water
5 '-phosphonucleaseInosine+single phosphoric acid
Inosine+water
Inosine nucleosidaseRibose+uridylic
Ribose+oxidized coenzyme+water
Ribose 1-desaturaseRibonic acid+reduced coenzyme
The end reaction thing is placed under the biochemical analyzer, detect the speed that predominant wavelength 340nm absorbancy rises, thereby calculate 5 '-the active size of phosphonuclease.
The reaction times of each reactions steps was controlled at 15 minutes.
Embodiment three (three doses)
5 of present embodiment '-the phosphonuclease diagnostic reagent is three doses, has:
Reagent I
Damping fluid 120mmol/l
Oxidized coenzyme 6mmo/l
Stablizer 20mmol/l
Reagent II
Damping fluid 120mmol/l
Inosine nucleosidase 10000U/l
Ribose 1-dehydrogenase 10 000U/l
Stablizer 50% (cumulative volume)
Reagent III
Damping fluid 120mmol/l
Inosine list phosphatase 11 0mmol/l
Stablizer 20mmol/l
On automatic clinical chemistry analyzer, set: 25 ℃ of temperature, 20 minutes reaction times, test predominant wavelength 340nm, more than the test commplementary wave length 405nm, tested 5 '-volume ratio of phosphonuclease sample and reagent is 1/25, the Direction of Reaction is positive reaction, 1 minute time of lag, 2 minutes detection times, theoretical k value-4180.
Concrete determination step is:
Inosine list phosphoric acid+water
5 '-phosphonucleaseInosine+single phosphoric acid
Inosine+water
Inosine nucleosidaseRibose+uridylic
Ribose+oxidized coenzyme+water
Ribose 1-desaturaseRibonic acid+reduced coenzyme
The end reaction thing is placed under the biochemical analyzer, detect the speed that predominant wavelength 340nm absorbancy rises, thereby calculate 5 '-the active size of phosphonuclease.
The reaction times of each reactions steps was controlled at 20 minutes.
Embodiment four
5 of present embodiment '-the phosphonuclease diagnostic reagent has:
Reagent I
Damping fluid 100mmol/l
Oxidized coenzyme 2mmo/l
Stablizer 20mmol/l
Reagent II
Damping fluid 100mmol/l
Inosine list phosphoric acid 3mmol/l
Inosine nucleosidase 6000U/l
Ribose 1-desaturase 6000U/l
Stablizer 50% (cumulative volume)
On Biochemical Analyzer, set: 37 ℃ of temperature, 10 minutes reaction times, test predominant wavelength 340nm, more than the test commplementary wave length 405nm, tested 5 '-volume ratio of phosphonuclease sample and reagent is 1/25, the Direction of Reaction is positive reaction, 1 minute time of lag, 2 minutes detection times, theoretical k value-4180.
After adding sample and reagent, make it to mix, following reaction take place:
Inosine list phosphoric acid+water
5 '-phosphonucleaseInosine+single phosphoric acid
Inosine+water
Inosine nucleosidaseRibose+uridylic
Ribose+oxidized coenzyme+water
Ribose 1-desaturaseRibonic acid+reduced coenzyme
The end reaction thing is placed under the biochemical analyzer, detect the speed that predominant wavelength 340nm absorbancy rises, thereby calculate 5 '-the active size of phosphonuclease.
In a word, experiment showed, and adopt measuring method of the present invention can draw required measurement result by ultraviolet analyser or half, automatic clinical chemistry analyzer device fully, and highly sensitive, tolerance range good, is not subjected to the pollution of inside and outside source material.
Claims (9)
1. one kind 5 '-the activity of 5 '-nucleotidase measuring method, step is as follows:
1), with sample and the reagent mix of mainly forming by inosine list phosphoric acid, oxidized coenzyme, Inosine nucleosidase, ribose 1-desaturase, make it to take place the reaction of following principle:
Inosine list phosphoric acid+water
5 '-phosphonucleaseInosine+single phosphoric acid
Inosine+water
Inosine nucleosidaseRibose+uridylic
Ribose+oxidized coenzyme+water
Ribose 1-desaturaseRibonic acid+reduced coenzyme
2), detect the end reaction thing in the speed that predominant wavelength 340nm absorbancy rises, calculate 5 '-the active size of phosphonuclease.
According to claim 1 described 5 '-the activity of 5 '-nucleotidase measuring method, it is characterized in that: described step 2) be, the end reaction thing is placed under ultraviolet analyser or half, the automatic clinical chemistry analyzer, detect the speed that predominant wavelength 340nm absorbancy rises, calculate 5 '-the active size of phosphonuclease.
3, according to claim 1 or 2 described mensuration 5 '-method of activity of 5 '-nucleotidase, it is characterized in that: temperature is controlled at 20 ℃ to 50 ℃ scopes, and the reaction times was controlled at 2-30 minute.
4, according to claim 1 or 2 described mensuration 5 '-method of activity of 5 '-nucleotidase, it is characterized in that: tested 5 '-ratio control of phosphonuclease sample and reagent is 1/10 to 1/250.
5. one kind 5 '-the phosphonuclease diagnostic kit, be grouped into by following one-tenth:
Damping fluid 40--200mmol/l
Inosine list phosphoric acid 0.5--50mmol/l
Oxidized coenzyme 0.5--10mmo/l
Inosine nucleosidase 500--50000U/l
Ribose 1-desaturase 500--50000U/l
Stablizer 10--80% (cumulative volume)
According to claim 5 described 5 '-the phosphonuclease diagnostic kit, it is characterized in that: described reagent is made into single agent, two agent or three doses.
According to claim 5 or 6 described 5 '-the phosphonuclease diagnostic kit, it is characterized in that: described stablizer is at least a in ethylene glycol, propylene glycol, glycerine, glycan, polyalcohols, ammonium sulfate or the salt.
According to claim 5 or 6 described 5 '-the phosphonuclease diagnostic kit, it is characterized in that: the pH scope of described buffer reagent is 6.0～11.0, and described buffer reagent is a kind of in three (carboxymethyl) aminomethane-hydrochloride buffer, trolamine damping fluid, 2-amino-2-methyl-1-propanol damping fluid, imidazole buffer or the glycylglycine damping fluid.
9, according to claim 5 or 6 described 5 '-the phosphonuclease diagnostic kit, it is characterized in that: described oxidized coenzyme is NADP
+, NAD
+Or thio-NAD
+A kind of in the oxidized form nicotinamide coenzyme or derivatives thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410065564 CN1778949A (en) | 2004-11-23 | 2004-11-23 | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410065564 CN1778949A (en) | 2004-11-23 | 2004-11-23 | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1778949A true CN1778949A (en) | 2006-05-31 |
Family
ID=36769404
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410065564 Pending CN1778949A (en) | 2004-11-23 | 2004-11-23 | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1778949A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109112181A (en) * | 2017-06-24 | 2019-01-01 | 浙江亚培生物技术有限公司 | A kind of 5`- activity of 5 '-nucleotidase measuring method and 5`- nucleotidase detection kit |
-
2004
- 2004-11-23 CN CN 200410065564 patent/CN1778949A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109112181A (en) * | 2017-06-24 | 2019-01-01 | 浙江亚培生物技术有限公司 | A kind of 5`- activity of 5 '-nucleotidase measuring method and 5`- nucleotidase detection kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1769481A (en) | Blood ammonia content determination method and blood ammonia diagnosis kit | |
| CN1778963A (en) | Determination of blood ammonia content and blood ammonia diagnostic reagent kit | |
| CN1778949A (en) | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase | |
| CN1778951A (en) | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'nucleotidase | |
| CN1778950A (en) | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase | |
| CN1778948A (en) | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase | |
| CN1302119C (en) | Method for measuring 5'-nucleotidase activity and diagnostic reagent kit of 5'-nucleotidase | |
| CN1778964A (en) | Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase | |
| CN1786692A (en) | Process for determining content of potassium ion by enzyme method and kit for diagnosing potassium ion thereof | |
| CN1778953A (en) | Determination of inorganic phosphorus and its diagnostic reagent kit | |
| CN1778937A (en) | Determination of blood ammonia content and blood ammonia diagnostic reagent kit | |
| CN1778943A (en) | Determination of inorganic phosphorus and its diagnostic reagent kit | |
| CN1778965A (en) | Determination of 5'-nucleotidase acitivity and its diagnostic reagent kit of 5'-nucleotidase | |
| CN1769473A (en) | Adenosine deaminase activity determination method and adenosine deaminase diagnosi kit | |
| CN1778955A (en) | Determination of inorganic phosphorus and its diagnostic reagent kit | |
| CN1778939A (en) | Creatinine content determination and creatinine diagnostic reagent kit | |
| CN1641041A (en) | Adenosin-deaminase diagnostic kit and adenosine deaminease activity determining method | |
| CN1778958A (en) | Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit | |
| CN1786187A (en) | Enzymatic method for determining potassium ion content and potassium ion diagnosis kit | |
| CN1778957A (en) | Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit | |
| CN1778941A (en) | Determination of inorganic phosphorus and its diagnostic reagent kit | |
| CN1778962A (en) | Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit | |
| CN1778956A (en) | Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit | |
| CN1778954A (en) | Determination of inorganic phosphorus and its diagnostic reagent kit | |
| CN1778959A (en) | Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |