CN114058676B - Factor Xa resisting activity determination kit - Google Patents
Factor Xa resisting activity determination kit Download PDFInfo
- Publication number
- CN114058676B CN114058676B CN202110748583.6A CN202110748583A CN114058676B CN 114058676 B CN114058676 B CN 114058676B CN 202110748583 A CN202110748583 A CN 202110748583A CN 114058676 B CN114058676 B CN 114058676B
- Authority
- CN
- China
- Prior art keywords
- factor
- reagent
- kit
- heparin
- buffer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000000694 effects Effects 0.000 title claims abstract description 31
- 108010074860 Factor Xa Proteins 0.000 title abstract description 26
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 38
- 239000007788 liquid Substances 0.000 claims abstract description 5
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 abstract description 19
- 229960002897 heparin Drugs 0.000 abstract description 19
- 229920000669 heparin Polymers 0.000 abstract description 19
- 239000000872 buffer Substances 0.000 abstract description 13
- 239000003593 chromogenic compound Substances 0.000 abstract description 9
- 239000003755 preservative agent Substances 0.000 abstract description 7
- 230000002335 preservative effect Effects 0.000 abstract description 7
- 239000004094 surface-active agent Substances 0.000 abstract description 6
- 229910017053 inorganic salt Inorganic materials 0.000 abstract description 5
- 238000003149 assay kit Methods 0.000 abstract description 2
- 229940051841 polyoxyethylene ether Drugs 0.000 description 13
- 229920000056 polyoxyethylene ether Polymers 0.000 description 13
- 239000003814 drug Substances 0.000 description 10
- -1 pentosan Chemical compound 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 229940079593 drug Drugs 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 229910052708 sodium Inorganic materials 0.000 description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 239000011734 sodium Substances 0.000 description 6
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 5
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 5
- 230000010100 anticoagulation Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 5
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical compound Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 230000015271 coagulation Effects 0.000 description 4
- 238000005345 coagulation Methods 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- TYMLOMAKGOJONV-UHFFFAOYSA-N 4-nitroaniline Chemical compound NC1=CC=C([N+]([O-])=O)C=C1 TYMLOMAKGOJONV-UHFFFAOYSA-N 0.000 description 3
- 229940123583 Factor Xa inhibitor Drugs 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 3
- AZFNGPAYDKGCRB-XCPIVNJJSA-M [(1s,2s)-2-amino-1,2-diphenylethyl]-(4-methylphenyl)sulfonylazanide;chlororuthenium(1+);1-methyl-4-propan-2-ylbenzene Chemical compound [Ru+]Cl.CC(C)C1=CC=C(C)C=C1.C1=CC(C)=CC=C1S(=O)(=O)[N-][C@@H](C=1C=CC=CC=1)[C@@H](N)C1=CC=CC=C1 AZFNGPAYDKGCRB-XCPIVNJJSA-M 0.000 description 3
- 230000001133 acceleration Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000003055 low molecular weight heparin Substances 0.000 description 3
- 229940127215 low-molecular weight heparin Drugs 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 235000010289 potassium nitrite Nutrition 0.000 description 3
- 239000004304 potassium nitrite Substances 0.000 description 3
- 229960001148 rivaroxaban Drugs 0.000 description 3
- KGFYHTZWPPHNLQ-AWEZNQCLSA-N rivaroxaban Chemical compound S1C(Cl)=CC=C1C(=O)NC[C@@H]1OC(=O)N(C=2C=CC(=CC=2)N2C(COCC2)=O)C1 KGFYHTZWPPHNLQ-AWEZNQCLSA-N 0.000 description 3
- 235000010288 sodium nitrite Nutrition 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- ZTQSAGDEMFDKMZ-UHFFFAOYSA-N Butyraldehyde Chemical compound CCCC=O ZTQSAGDEMFDKMZ-UHFFFAOYSA-N 0.000 description 2
- 206010053567 Coagulopathies Diseases 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- 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 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- 229920001499 Heparinoid Polymers 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 150000005215 alkyl ethers Chemical class 0.000 description 2
- 239000003146 anticoagulant agent Substances 0.000 description 2
- 229940127219 anticoagulant drug Drugs 0.000 description 2
- 239000004019 antithrombin Substances 0.000 description 2
- 208000034158 bleeding Diseases 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 150000002191 fatty alcohols Chemical class 0.000 description 2
- 239000002554 heparinoid Substances 0.000 description 2
- 229940025770 heparinoids Drugs 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000002452 interceptive effect Effects 0.000 description 2
- GKQWYZBANWAFMQ-UHFFFAOYSA-M lithium;2-hydroxypropanoate Chemical compound [Li+].CC(O)C([O-])=O GKQWYZBANWAFMQ-UHFFFAOYSA-M 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- PGOHTUIFYSHAQG-LJSDBVFPSA-N (2S)-6-amino-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-1-[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-4-methylsulfanylbutanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-carbamimidamidopentanoyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]-3-methylbutanoyl]amino]-4-methylpentanoyl]amino]-4-methylpentanoyl]amino]acetyl]amino]-3-hydroxypropanoyl]amino]-4-methylpentanoyl]amino]-3-sulfanylpropanoyl]amino]-4-methylsulfanylbutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-hydroxybutanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-hydroxypropanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1H-imidazol-5-yl)propanoyl]amino]-4-methylpentanoyl]amino]-3-hydroxybutanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-oxopentanoyl]amino]-3-hydroxybutanoyl]amino]-3-hydroxypropanoyl]amino]-3-carboxypropanoyl]amino]-3-hydroxypropanoyl]amino]-5-oxopentanoyl]amino]-5-oxopentanoyl]amino]-3-phenylpropanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-methylbutanoyl]amino]-4-methylpentanoyl]amino]-4-oxobutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-4-carboxybutanoyl]amino]-5-oxopentanoyl]amino]hexanoic acid Chemical compound CSCC[C@H](N)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O PGOHTUIFYSHAQG-LJSDBVFPSA-N 0.000 description 1
- ZYECEUZMVSGTMR-LBDWYMBGSA-N (4s)-4-[[(2s,3s)-2-benzamido-3-methylpentanoyl]amino]-5-[[2-[[(2s)-5-(diaminomethylideneamino)-1-(4-nitroanilino)-1-oxopentan-2-yl]amino]-2-oxoethyl]amino]-5-oxopentanoic acid Chemical compound N([C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CCCN=C(N)N)C(=O)NC=1C=CC(=CC=1)[N+]([O-])=O)C(=O)C1=CC=CC=C1 ZYECEUZMVSGTMR-LBDWYMBGSA-N 0.000 description 1
- 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
- 125000000954 2-hydroxyethyl group Chemical group [H]C([*])([H])C([H])([H])O[H] 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- ZNBNBTIDJSKEAM-UHFFFAOYSA-N 4-[7-hydroxy-2-[5-[5-[6-hydroxy-6-(hydroxymethyl)-3,5-dimethyloxan-2-yl]-3-methyloxolan-2-yl]-5-methyloxolan-2-yl]-2,8-dimethyl-1,10-dioxaspiro[4.5]decan-9-yl]-2-methyl-3-propanoyloxypentanoic acid Chemical compound C1C(O)C(C)C(C(C)C(OC(=O)CC)C(C)C(O)=O)OC11OC(C)(C2OC(C)(CC2)C2C(CC(O2)C2C(CC(C)C(O)(CO)O2)C)C)CC1 ZNBNBTIDJSKEAM-UHFFFAOYSA-N 0.000 description 1
- QNZCBYKSOIHPEH-UHFFFAOYSA-N Apixaban Chemical compound C1=CC(OC)=CC=C1N1C(C(=O)N(CC2)C=3C=CC(=CC=3)N3C(CCCC3)=O)=C2C(C(N)=O)=N1 QNZCBYKSOIHPEH-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- HGVDHZBSSITLCT-JLJPHGGASA-N Edoxaban Chemical compound N([C@H]1CC[C@@H](C[C@H]1NC(=O)C=1SC=2CN(C)CCC=2N=1)C(=O)N(C)C)C(=O)C(=O)NC1=CC=C(Cl)C=N1 HGVDHZBSSITLCT-JLJPHGGASA-N 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- YHSHAHLOHLZILM-UHFFFAOYSA-L S(=S)(=O)([O-])[O-].[Na+].C(C)[Hg].[Na+] Chemical compound S(=S)(=O)([O-])[O-].[Na+].C(C)[Hg].[Na+] YHSHAHLOHLZILM-UHFFFAOYSA-L 0.000 description 1
- 108010022999 Serine Proteases Proteins 0.000 description 1
- 102000012479 Serine Proteases Human genes 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 108010000499 Thromboplastin Proteins 0.000 description 1
- 102000002262 Thromboplastin Human genes 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 206010047249 Venous thrombosis Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 150000003973 alkyl amines Chemical class 0.000 description 1
- 230000001858 anti-Xa Effects 0.000 description 1
- 229960003886 apixaban Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000007982 barbital buffer Substances 0.000 description 1
- 108010031969 benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide Proteins 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 208000015294 blood coagulation disease Diseases 0.000 description 1
- 230000002612 cardiopulmonary effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 230000035602 clotting Effects 0.000 description 1
- 230000009852 coagulant defect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- XEKSTYNIJLDDAZ-JASSWCPGSA-D decasodium;(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5r,6r)-6-[(2r,3s,4s,5r,6r)-2-carboxylato-4-hydroxy-6-[(2r,3s,4r,5r,6s)-4-hydroxy-6-methoxy-5-(sulfonatoamino)-2-(sulfonatooxymethyl)oxan-3-yl]oxy-5-sulfonatooxyoxan-3-yl]oxy-5-(sulfonatoamino)-4-sulfonatooxy-2-(sul Chemical compound [Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].O[C@@H]1[C@@H](NS([O-])(=O)=O)[C@@H](OC)O[C@H](COS([O-])(=O)=O)[C@H]1O[C@H]1[C@H](OS([O-])(=O)=O)[C@@H](O)[C@H](O[C@@H]2[C@@H]([C@@H](OS([O-])(=O)=O)[C@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O[C@@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](COS([O-])(=O)=O)O4)NS([O-])(=O)=O)[C@H](O3)C([O-])=O)O)[C@@H](COS([O-])(=O)=O)O2)NS([O-])(=O)=O)[C@H](C([O-])=O)O1 XEKSTYNIJLDDAZ-JASSWCPGSA-D 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 229960000622 edoxaban Drugs 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229960003661 fondaparinux sodium Drugs 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- 229940049906 glutamate Drugs 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 229910000378 hydroxylammonium sulfate Inorganic materials 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229940028435 intralipid Drugs 0.000 description 1
- 239000002960 lipid emulsion Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 235000013923 monosodium glutamate Nutrition 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- KDFYOQXKHPVLKX-UHFFFAOYSA-M sodium 3-[bis(2-hydroxyethyl)amino]-2-hydroxypropane-1-sulfonic acid hydroxide Chemical compound [OH-].[Na+].OCCN(CCO)CC(CS(=O)(=O)O)O KDFYOQXKHPVLKX-UHFFFAOYSA-M 0.000 description 1
- 229940073490 sodium glutamate Drugs 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- QBQVXXQXZXDEHE-UHFFFAOYSA-M sodium;propane-1-sulfonate;hydrate Chemical compound O.[Na+].CCCS([O-])(=O)=O QBQVXXQXZXDEHE-UHFFFAOYSA-M 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 229940033663 thimerosal Drugs 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 206010043554 thrombocytopenia Diseases 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/56—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Neurosurgery (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The application relates to an anti-factor Xa activity assay kit. The kit comprises a first reagent and a second reagent; the first reagent comprises a chromogenic substrate, a buffer, a surfactant, a preservative, and the second reagent comprises factor Xa, a buffer, an inorganic salt, a preservative. Compared with the existing heparin determination reagent, the reagent kit can ensure the stability and anti-interference performance of the liquid reagent.
Description
Technical Field
The application relates to the fields of medicine, immunity and in-vitro diagnosis. Specifically, the application provides a kit for detecting anti-factor Xa activity in a sample.
Background
Factor Xa is a serine protease that is located upstream of the cascade of blood coagulation reactions. Factor Xa is well centered in the intrinsic and extrinsic coagulation pathways. Factor Xa inhibitors therefore can block both endogenous and exogenous coagulation.
Factor Xa is known to be inhibited by various drugs such as heparin, pentosan, rivaroxaban Rivaroxaban, apixaban, edoxaban. The same feature of these drugs is that they all lead to inhibition of factor Xa.
Heparin is used as the most commonly used factor Xa inhibitor, and the main mechanism of action is to enhance the inhibition of thrombin by antithrombin (about 2000-fold improvement).
According to different molecular weights, heparin drugs can be classified into common heparin, low molecular weight heparin and synthetic heparin (such as fondaparinux sodium, rivaroxaban, etc.). Common heparin can inhibit factors Xa and IIa with the same efficacy to play an anticoagulant role; the low molecular weight heparin inhibits factor Xa more strongly; the artificially synthesized heparinoids and oral anticoagulation heparinoids only have the effect of resisting factor Xa, so that anticoagulation is controlled, and side reactions such as bleeding and the like can be avoided.
Monitoring of the drug dose of factor Xa inhibitors (e.g. heparin) is necessary. Especially, when treating acute venous thrombosis, the treatment effect of heparin is ensured, and the risk of bleeding is reduced. Common heparin treatment is typically performed in hospitals and monitored by Activated Partial Thromboplastin Time (APTT) or anti-Xa factor. High dose plain heparin therapy is commonly used in surgery (e.g., cardiopulmonary bypass) and efficacy is monitored by Activated Clotting Time (ACT). Low molecular weight heparin treatment is typically outpatient or inpatient, and conventional treatment does not require monitoring, and if desired, heparin efficacy can be observed by anti-factor Xa.
Currently, coagulation and chromogenic substrate methods are known to detect the activity against factor Xa. The detection of the anti-factor Xa by the chromogenic substrate method directly judges the effect of inhibiting the factor Xa by the medicament, has accurate result, and reflects the real anticoagulation effect of the anticoagulation medicament (directly or indirectly inhibiting the factor Xa) in the patient. The principle is as follows: the complex formed by the anticoagulant drug and AT in the sample or the factor Xa which is excessively added in the direct inhibition factor Xa drug inhibitor. The residual activity of the residual factor Xa is measured, and the result of the residual activity is inversely proportional to the effective concentration of the drug in the sample, so that the actual anticoagulation effect of the drug is estimated and expressed in the unit of anti-factor Xa. The method is more specific than the traditional ACT and APTT experimental means, and is not interfered by the coagulation disorder, thrombocytopenia or blood dilution of the patient.
At present, the heparin activity is mostly measured by detecting the anti-factor Xa activity by using a chromogenic substrate method at home and abroad. Heparin (or other similar substances) in the plasma forms a complex with antithrombin, inhibiting the excessive addition of factor Xa; the remaining factor Xa acts with its specific substrate, and the absorbance of p-nitroaniline (pNA) produced is detected at 405nm, the pNA chromogenic amount being inversely proportional to the heparin (or other similar substance) concentration in the plasma to be measured. For example, CN103063592a discloses a method for measuring heparin activity, and a method for measuring heparin activity with high sensitivity and accuracy is established.
However, the liquid reagents of the chromogenic substrate method are poor in stability and only freeze-dried preservation reagents are available; or the reagent has poor anti-interference performance, and is inaccurate for the measurement of hemolysis, lipidemia and hyperbilirubin samples with high frequency in clinical test samples.
In view of this, CN108195783a discloses a heparin activity assay kit, which finds that adding a reducing agent (such as hydroxylamine hydrochloride, polyoxyethylene alkyl ether) to a first reagent of the reagent can improve the anti-hemoglobin interference effect, while polyoxyethylene dodecyl ether can improve the anti-chyle interference effect, and a stabilizing agent (such as mannitol, sorbitol, polyethylene glycol) can effectively improve the stability of a chromogenic substrate. The combination of glycine, lithium lactate and sodium glutamate can effectively stabilize the potency of Xa factor.
Achieving uniformity of stability and tamper resistance of liquid reagents is a known problem in the art.
Disclosure of Invention
According to some embodiments, a kit for determining anti-factor Xa activity is provided comprising a first reagent and a second reagent.
In the present application, "first" and "second" do not limit the order or position of elements, compositions, elements, features, and are used only to distinguish between different elements, compositions, elements, features.
In some embodiments, the first agent comprises a chromogenic substrate, a buffer, a surfactant, a preservative.
In some embodiments, the second agent comprises factor Xa, a buffer, an inorganic salt, a preservative.
In some embodiments, the buffers in the first and second reagents are the same or different (in composition and/or concentration).
In some embodiments, the preservative in the first and second agents are the same or different (in composition and/or concentration).
In some embodiments, the buffer is selected from any one or a combination of the following: 3- [ N, N-bis (hydroxyethyl) amino ] -2-hydroxypropanesulfonic acid-sodium hydroxide buffer, 4- (2-hydroxyethyl) -1-piperazine propanesulfonic acid-sodium hydroxide buffer, tris-hydroxymethyl aminomethane-HCl buffer, phosphate buffer, glycine buffer, barbital buffer.
In some embodiments, the concentration of the buffer is 50mM to 500mM; for example, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500mM.
In some embodiments, the buffer in the first reagent and the buffer in the second reagent is 50mM Tris buffer.
In some embodiments, the chromogenic substrate is selected from any one or a combination of the following: bz-Ile-Glu-Gly-Arg-pNA, suc-Ile-Glu (gamma-pip) -Gly-Arg-pNA.HCl, N-alpha-Z-D-Arg-Gly-Arg-pNA.2HCl, Z-D-Arg-Gly-Arg-pNA.2HCl.
In some embodiments, the concentration of the chromogenic substrate is 0.5mM to 2.5mM; for example 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5mM.
In some embodiments, the kit for determining anti-factor Xa activity does not comprise any one or a combination selected from the group consisting of: hydroxylamine hydrochloride, hydroxylamine sulfate, n-butyraldehyde, tartaric acid, ascorbic acid, formic acid, mannitol, sorbitol, polyethylene glycol, glycine, lithium lactate, glutamate.
In some embodiments, the first reagent does not comprise sodium nitrite and/or potassium nitrite.
In some embodiments, the preservative is selected from any one or a combination of the following: sodium azide, thimerosal, phenol, ethyl mercury sodium thiosulfate.
In some embodiments, the concentration of the preservative is w/v 0.05% to 0.1%, e.g., 0.05, 0.06, 0.07, 0.08, 0.09, 0.1%.
In some embodiments, the surfactant is selected from any one or a combination of the following: fatty alcohol polyoxyethylene ether, sodium fatty alcohol polyoxyethylene ether sulfate, polyoxyethylene alkyl ether, polyoxyethylene alkylamine, and polyoxyethylene sorbitan fatty acid ester.
In some embodiments, the first reagent comprises: and a combination of fatty alcohol-polyoxyethylene ether and sodium fatty alcohol-polyoxyethylene ether sulfate.
In some embodiments, the concentration of the surfactant is from 0.2% to 0.6% w/v; for example, 0.2, 0.3, 0.4, 0.5, 0.6%.
In some embodiments, the first reagent comprises a mass ratio of 3:1 and sodium fatty alcohol-polyoxyethylene ether sulfate.
In some embodiments, the inorganic salt is selected from any one or a combination of the following: sodium nitrite and potassium nitrite are preferred.
In some embodiments, the concentration of inorganic salt is from 0.1% to 0.5%, e.g., 0.1, 0.2, 0.3, 0.4, 0.5% w/v.
In some embodiments, factor Xa is selected from any one or a combination of the following: bovine factor Xa, human factor Xa, porcine factor Xa, preferably bovine factor Xa.
In some embodiments, the concentration of factor Xa is from 0.4IU/mL to 1IU/mL, such as 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0IU/mL.
In a specific embodiment, a kit for determining anti-factor Xa activity is provided, consisting of:
1) A first reagent;
2) A second reagent;
the first reagent consists of:
the second reagent consists of:
in a specific embodiment, the first reagent and the second reagent are liquid reagents.
In some embodiments, there is provided the use of a combination of fatty alcohol-polyoxyethylene ether and sodium fatty alcohol-polyoxyethylene ether sulfate to improve the stability or interference resistance of a kit, particularly a kit for quantifying anti-factor Xa activity.
In the present application, anti-factor Xa activity refers to any known or future agent (or inhibitor) that is capable of inhibiting the activity of factor Xa and that can act anywhere in the factor Xa-associated pathway in the endogenous or exogenous coagulation pathway, such an effect being permitted either directly or indirectly.
In some embodiments, there is provided the use of nitrite (preferably sodium/potassium nitrite) to improve the stability or interference resistance of a kit, particularly a kit for quantifying anti-factor Xa activity.
In some embodiments, there is provided the use of a combination of nitrite, fatty alcohol polyoxyethylene ether and sodium fatty alcohol polyoxyethylene ether sulfate to improve the stability or interference resistance of a kit, particularly a kit for quantifying anti-factor Xa activity.
In some embodiments, the nitrite is located in a different container than the fatty alcohol-polyoxyethylene ether and the sodium fatty alcohol-polyoxyethylene ether sulfate during the preparation and storage of the kit.
In some embodiments, the mass ratio of nitrite, fatty alcohol-polyoxyethylene ether, and fatty alcohol-polyoxyethylene ether sodium sulfate is 3:6:2.
Detailed Description
Example 1: preparation of the kit and the control kit of the application
In order to verify the stability and anti-interference effect of the kit, the kit is prepared with kits with different surfactant and inorganic salt adding modes, and the formula is as follows:
TABLE 1 preparation of the kit (A) according to the application
TABLE 2 preparation of control kit (B)
TABLE 3 preparation of control kit (C)
TABLE 4 preparation of control kit (D)
TABLE 5 preparation of control kit (E)
TABLE 6 preparation of control kit (F)
Example 2: stability test of the inventive kit and control kit
The inventive kit a and the control kits B to F prepared according to example 1 were placed at 37 degrees for different days for thermal acceleration destruction and then taken out, and absorbance change rate detection was performed with the conventional cryopreserved kit. The detection sample is heparin calibrator with theoretical concentration of 0, 0.8 and 2.0 IU/ml.
The detection instrument may be selected from various types of coagulometer such as STA R-MAX instrument of Stago, CS5100 instrument of hssenmeikang, TOP750 instrument of us Wo Fen, etc. The specific parameters can be adjusted appropriately according to different instruments, and the detection data of the instrument of the Hissenkang CS5100 are as follows:
TABLE 7 acceleration stability results
It can be seen from the comparison that changing the nitrite in the second reagent significantly affects the stability of the reagent.
Example 3: anti-interference test of the kit and the control kit
The recovery rate was calculated by comparing samples of the present application (containing 100mg/dL, 300mg/dL, 500mg/dL human hemoglobin Hb, 5mg/dL, 10mg/dL, 15mg/dL, 20mg/dL bilirubin DB, 100mg/dL, 200mg/dL, 300mg/dL, 400mg/dL fat emulsion Intralipid) prepared in example 1 and samples containing no interfering substance at different concentrations with the samples containing no interfering substance, and the detection results were as follows:
TABLE 8 anti-interference test
As can be seen by comparison, the surfactant replacement of the first reagent results in a poor tamper resistance.
In summary, the specific composition of the kit a of the present application is superior in both stability and anti-interference properties in combination with data of acceleration stability and anti-interference.
Claims (2)
1. A kit for determining anti-factor Xa activity, consisting of:
1) A first reagent;
2) A second reagent;
the first reagent consists of:
the second reagent consists of:
2. the kit for determining anti-factor Xa activity according to claim 1, wherein:
the first reagent and the second reagent are liquid reagents.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2020107580041 | 2020-07-31 | ||
| CN202010758004 | 2020-07-31 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114058676A CN114058676A (en) | 2022-02-18 |
| CN114058676B true CN114058676B (en) | 2023-12-01 |
Family
ID=80233283
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110748583.6A Active CN114058676B (en) | 2020-07-31 | 2021-07-02 | Factor Xa resisting activity determination kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114058676B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102507957A (en) * | 2011-11-01 | 2012-06-20 | 北京利德曼生化股份有限公司 | Method and kit for determining apolipoprotein A2 by using immunity transmission turbidimetric method |
| CN103063592A (en) * | 2012-12-31 | 2013-04-24 | 中国医学科学院输血研究所 | Determination method for heparin activity |
| CN104048931A (en) * | 2014-04-21 | 2014-09-17 | 上海贞元诊断用品科技有限公司 | Heparin content detection method |
| CN104062243A (en) * | 2014-04-21 | 2014-09-24 | 上海贞元诊断用品科技有限公司 | FXa activity detection reagent, preparation method and application of FXa activity detection reagent |
| CN105548574A (en) * | 2016-02-02 | 2016-05-04 | 潍坊三维生物工程集团有限公司 | Kit and method for detecting content of C1 inhibitor and application |
| AU2016203992A1 (en) * | 2009-05-11 | 2016-06-30 | Berg Llc | Methods for the diagnosis of metabolic disorders using epimetabolic shifters, multidimensional intracellular molecules, or environmental influencers |
| CN108195783A (en) * | 2018-01-30 | 2018-06-22 | 迈克生物股份有限公司 | Heparin activity determination kit |
| CN109991177A (en) * | 2017-12-30 | 2019-07-09 | 济南宇鑫生物科技有限公司 | A kind of stabilization, bilirubin direct (enzymatic measurement) detection reagent of strong antijamming capability and detection method |
| CN110261625A (en) * | 2019-07-15 | 2019-09-20 | 三诺生物传感股份有限公司 | A kind of bilirubin multiplexed detection reagents box and its application method |
-
2021
- 2021-07-02 CN CN202110748583.6A patent/CN114058676B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2016203992A1 (en) * | 2009-05-11 | 2016-06-30 | Berg Llc | Methods for the diagnosis of metabolic disorders using epimetabolic shifters, multidimensional intracellular molecules, or environmental influencers |
| CN102507957A (en) * | 2011-11-01 | 2012-06-20 | 北京利德曼生化股份有限公司 | Method and kit for determining apolipoprotein A2 by using immunity transmission turbidimetric method |
| CN103063592A (en) * | 2012-12-31 | 2013-04-24 | 中国医学科学院输血研究所 | Determination method for heparin activity |
| CN104048931A (en) * | 2014-04-21 | 2014-09-17 | 上海贞元诊断用品科技有限公司 | Heparin content detection method |
| CN104062243A (en) * | 2014-04-21 | 2014-09-24 | 上海贞元诊断用品科技有限公司 | FXa activity detection reagent, preparation method and application of FXa activity detection reagent |
| CN105548574A (en) * | 2016-02-02 | 2016-05-04 | 潍坊三维生物工程集团有限公司 | Kit and method for detecting content of C1 inhibitor and application |
| CN109991177A (en) * | 2017-12-30 | 2019-07-09 | 济南宇鑫生物科技有限公司 | A kind of stabilization, bilirubin direct (enzymatic measurement) detection reagent of strong antijamming capability and detection method |
| CN108195783A (en) * | 2018-01-30 | 2018-06-22 | 迈克生物股份有限公司 | Heparin activity determination kit |
| CN110261625A (en) * | 2019-07-15 | 2019-09-20 | 三诺生物传感股份有限公司 | A kind of bilirubin multiplexed detection reagents box and its application method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114058676A (en) | 2022-02-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Haynes et al. | Elevated fibrin (ogen) degradation products in the adult respiratory distress syndrome | |
| Esposito et al. | The role of the activated clotting time in heparin administration and neutralization for cardiopulmonary bypass | |
| EP0525035B1 (en) | Coagulation assays and reagents | |
| Kumada et al. | Comparative study on heparin and a synthetic thrombin inhibitor No. 805 (MD-805) in experimental antithrombin III-deficient animals | |
| CA2392350A1 (en) | Hematological assay and reagent | |
| CN108195783A (en) | Heparin activity determination kit | |
| JP2002214238A (en) | Method for testing activated clotting time low in sensitivity to presence of aprotinin and method for evaluating adrotinin sensitivity | |
| Martindale et al. | The activated coagulation time: suitability for monitoring heparin effect and neutralization during pediatric cardiac surgery | |
| CN114277089A (en) | Dabigatran detection reagent and kit | |
| EP0927767A2 (en) | Improved thrombin-based assay for antithrombin-III | |
| CN109082458B (en) | Kit for quantitatively detecting oral blood coagulation factor Xa inhibitor by using thrombus elastography method and preparation method of kit | |
| CN114058676B (en) | Factor Xa resisting activity determination kit | |
| HOLM et al. | The antithrombotic effect of heparin in deep venous thrombosis: relation to four heparin assays | |
| US20150185236A1 (en) | Blood coagulation time prolonging agent | |
| EP0576038B1 (en) | Method for measuring tissue plasminogen activator, antithrombin III and soluble fibrin | |
| EP0259463B1 (en) | Heparin assay | |
| US20140155498A1 (en) | Method of Manufacture of Stable Liquid Coagulation Factors | |
| CN115586179A (en) | Kit for determining activity of antithrombin III and application thereof | |
| Danielsson et al. | Is there a need for antithrombin III substitution early after burn injury? | |
| US20190302132A1 (en) | Identification of anticoagulants in a sample | |
| Šaboviã et al. | The effect of long term, low-dose tranexamic acid treatment on platelet dysfunction and haemoglobin levels in haemodialysis patients | |
| JP6278454B2 (en) | Reagent and method for measuring direct thrombin inhibitor in biological sample | |
| Hoffmann et al. | Plasma heparin: a simple, one-sample determination of both effect and concentration based on the activated partial thromboplastin time | |
| US5114845A (en) | Assays for plasminogen activator inhibitor and soluble fibrin | |
| Scully | The use of an automated analyzer in the evaluation of antithrombin III and heparin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |