CN103333945A - Direct bilirubin detection reagent - Google Patents
Direct bilirubin detection reagent Download PDFInfo
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- CN103333945A CN103333945A CN2013102026622A CN201310202662A CN103333945A CN 103333945 A CN103333945 A CN 103333945A CN 2013102026622 A CN2013102026622 A CN 2013102026622A CN 201310202662 A CN201310202662 A CN 201310202662A CN 103333945 A CN103333945 A CN 103333945A
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- damping fluid
- acid
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- direct detection
- sodium
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- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 57
- 238000001514 detection method Methods 0.000 title claims abstract description 31
- 238000008789 Direct Bilirubin Methods 0.000 title abstract description 5
- 238000006243 chemical reaction Methods 0.000 claims abstract description 24
- 239000003085 diluting agent Substances 0.000 claims abstract description 20
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 14
- LCPVQAHEFVXVKT-UHFFFAOYSA-N 2-(2,4-difluorophenoxy)pyridin-3-amine Chemical compound NC1=CC=CN=C1OC1=CC=C(F)C=C1F LCPVQAHEFVXVKT-UHFFFAOYSA-N 0.000 claims abstract description 8
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Substances [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 claims abstract description 8
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims abstract description 7
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims abstract description 7
- DBVJJBKOTRCVKF-UHFFFAOYSA-N Etidronic acid Chemical compound OP(=O)(O)C(O)(C)P(O)(O)=O DBVJJBKOTRCVKF-UHFFFAOYSA-N 0.000 claims abstract description 7
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims abstract description 7
- 239000000872 buffer Substances 0.000 claims abstract description 5
- 239000012530 fluid Substances 0.000 claims description 50
- 238000013016 damping Methods 0.000 claims description 46
- 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 claims description 38
- 239000000463 material Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 6
- VGYYSIDKAKXZEE-UHFFFAOYSA-L hydroxylammonium sulfate Chemical compound O[NH3+].O[NH3+].[O-]S([O-])(=O)=O VGYYSIDKAKXZEE-UHFFFAOYSA-L 0.000 claims description 6
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical class COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 claims description 6
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Natural products OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 5
- 102000004316 Oxidoreductases Human genes 0.000 claims description 5
- 108090000854 Oxidoreductases Proteins 0.000 claims description 5
- 235000010323 ascorbic acid Nutrition 0.000 claims description 5
- 239000011668 ascorbic acid Substances 0.000 claims description 5
- 229960005070 ascorbic acid Drugs 0.000 claims description 5
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical class [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 claims description 5
- IHPYMWDTONKSCO-UHFFFAOYSA-N 2,2'-piperazine-1,4-diylbisethanesulfonic acid Chemical compound OS(=O)(=O)CCN1CCN(CCS(O)(=O)=O)CC1 IHPYMWDTONKSCO-UHFFFAOYSA-N 0.000 claims description 4
- 239000013543 active substance Substances 0.000 claims description 4
- SUMDYPCJJOFFON-UHFFFAOYSA-N isethionic acid Chemical compound OCCS(O)(=O)=O SUMDYPCJJOFFON-UHFFFAOYSA-N 0.000 claims description 4
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- 229920000136 polysorbate Polymers 0.000 claims description 4
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- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 4
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 claims description 4
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 4
- 229960005486 vaccine Drugs 0.000 claims description 4
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 3
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 claims description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 3
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- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
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- 239000007853 buffer solution Substances 0.000 claims description 3
- QDPMLKBAQOZXEF-UHFFFAOYSA-N ethanesulfonic acid;sodium Chemical compound [Na].CCS(O)(=O)=O QDPMLKBAQOZXEF-UHFFFAOYSA-N 0.000 claims description 3
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 claims description 3
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- 229940069338 potassium sorbate Drugs 0.000 claims description 3
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 claims description 3
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- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 3
- -1 (2-hydroxyethyl) amino Chemical group 0.000 claims description 2
- CKQPAVCUJDZLLE-UHFFFAOYSA-N 1,3-dihydroxy-2-(methylamino)butane-2-sulfonic acid Chemical compound CNC(CO)(C(C)O)S(O)(=O)=O CKQPAVCUJDZLLE-UHFFFAOYSA-N 0.000 claims description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims description 2
- UPMBDJWMINKVIJ-UHFFFAOYSA-N 2-amino-3-hydroxy-2-methylpropane-1-sulfonic acid Chemical compound OCC(N)(C)CS(O)(=O)=O UPMBDJWMINKVIJ-UHFFFAOYSA-N 0.000 claims description 2
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 claims description 2
- QFOHBWFCKVYLES-UHFFFAOYSA-N Butylparaben Chemical compound CCCCOC(=O)C1=CC=C(O)C=C1 QFOHBWFCKVYLES-UHFFFAOYSA-N 0.000 claims description 2
- OWXMKDGYPWMGEB-UHFFFAOYSA-N HEPPS Chemical compound OCCN1CCN(CCCS(O)(=O)=O)CC1 OWXMKDGYPWMGEB-UHFFFAOYSA-N 0.000 claims description 2
- 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 claims description 2
- XPJVKCRENWUEJH-UHFFFAOYSA-N Isobutylparaben Chemical compound CC(C)COC(=O)C1=CC=C(O)C=C1 XPJVKCRENWUEJH-UHFFFAOYSA-N 0.000 claims description 2
- CMHMMKSPYOOVGI-UHFFFAOYSA-N Isopropylparaben Chemical compound CC(C)OC(=O)C1=CC=C(O)C=C1 CMHMMKSPYOOVGI-UHFFFAOYSA-N 0.000 claims description 2
- 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 claims description 2
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- 239000002253 acid Substances 0.000 claims description 2
- CCIVGXIOQKPBKL-UHFFFAOYSA-N ethanesulfonic acid Chemical compound CCS(O)(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-N 0.000 claims description 2
- 235000010228 ethyl p-hydroxybenzoate Nutrition 0.000 claims description 2
- 239000004403 ethyl p-hydroxybenzoate Substances 0.000 claims description 2
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- NUVBSKCKDOMJSU-UHFFFAOYSA-N ethylparaben Chemical compound CCOC(=O)C1=CC=C(O)C=C1 NUVBSKCKDOMJSU-UHFFFAOYSA-N 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 claims description 2
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 claims description 2
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- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
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- CIJQGPVMMRXSQW-UHFFFAOYSA-M sodium;2-aminoacetic acid;hydroxide Chemical compound O.[Na+].NCC([O-])=O CIJQGPVMMRXSQW-UHFFFAOYSA-M 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical class OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims 2
- NHQAUSOKYWFWHO-UHFFFAOYSA-N 4-aminobutane-2-sulfonic acid Chemical compound OS(=O)(=O)C(C)CCN NHQAUSOKYWFWHO-UHFFFAOYSA-N 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 7
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- 108010024957 Ascorbate Oxidase Proteins 0.000 abstract 1
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- NXPHCVPFHOVZBC-UHFFFAOYSA-N hydroxylamine;sulfuric acid Chemical compound ON.OS(O)(=O)=O NXPHCVPFHOVZBC-UHFFFAOYSA-N 0.000 abstract 1
- 239000003223 protective agent Substances 0.000 abstract 1
- 235000002639 sodium chloride Nutrition 0.000 abstract 1
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Abstract
The invention discloses a direct bilirubin detection reagent. The direct bilirubin detection reagent comprises diluents and reaction reagents. The diluents comprise a buffer 1, a surfactant, an antiseptic and ascorbate oxidase. The reaction reagents comprise a buffer 2, common salt, disodium ethylene diamine tetraacetate, hydroxylamine sulphate, etidronic acid, sodium persulfate, sulfuric acid, an antiseptic and a freeze-drying protective agent. The direct bilirubin detection reagent has good sensitivity, accuracy, precision and linearity and can satisfy clinical examination requirements.
Description
Technical field
The present invention relates to the medical test technical field, be specifically related to a kind of bilirubin direct detection reagent for the POCT analyser.
Background technology
Bilirubin direct (Direct bilirubin, DBIL) claim conjugative bilirubin again, unconjugated bilirubin transforms in liver cell, be combined with glucuronic acid and form conjugative bilirubin, measure the type that DBIL is mainly used in differentiating jaundice, serum DBIL raises, and illustrates that bilirubin is from the drainage generation obstacle of biliary tract after liver cell is handled and handles, DBIL is one of project of medical test routine, occupies important status in class medicals diagnosis on disease such as liver and gall.DBIL generally adopts various large automatic Biochemical Analyzers to detect at present, but because large automatic Biochemical Analyzer equipment price height, and complicated operation, operator need have relevant expertise and accept corresponding training, and it is high to use complementary conditions to require, and need to be equipped with voltage stabilized source, water purification machine etc., and floor space is big, the maintenance cost height needs professional's time-based maintenance, so basic medical unit or household person all do not have condition to buy and use.In addition, the large hospital patient is many, detects that formality is loaded down with trivial details, long flow path, waiting time be long, and this has also brought the huge time to bear to the patient, the more important thing is that patient is affected adversely treatment because can not in time diagnosing, even can lose one's life.
(point-of-care testing is an emerging technical field POCT) to real-time test, and principal feature is to obtain a result fast, and is easy and simple to handle, uses easily and miniaturization.Along with diagnosis and the progress of ancillary technique, and people are to the understanding of disease and the requirement for the treatment of level raising, and POCT receives publicity gradually.In fact POCT becomes the important research project of external diagnosis reagent and instrument field gradually in American-European market, existing many products put goods on the market, but this class POCT instrument has a major defect at present, be exactly that analyser and reagent consumptive material are expensive especially, has huge primary care system like this for China, and the reagent usage quantity is country greatly, and instrument and the matched reagent consumptive material of American-European exploitation obviously all face the challenge at present.
The micro-fluidic chip technology is one of of paramount importance cutting edge technology in the 21 century world, it is integrated into basic operation units such as specimen preparation related in the fields such as biological and chemical, reaction, separation, detection and cell cultures, sorting, cracking on the chip of one tens square centimeters (even littler), form network by the microchannel, running through total system with controlled fluid, is a kind of technology in order to the various functions that replace conventional biological or chemical laboratory.The micro-fluidic chip technology is incorporated into POCT equipment, started the new situation of POCT development, can make the complicated whole blood in laboratory in the past quantitatively, blood cell serum separates, serum dilution and step such as mensuration synchronously, finishes in the on-line automatic equalization of chip, reaches multiple mark synchronous detection purpose.POCT analyser in conjunction with the micro-fluidic chip technology has split hair caccuracy, low blood volume, simple to operate, few, the low cost and other advantages of detection reagent consumption of needing concurrently, as protect and give birth to the international POCT instrument of curing company limited of giving birth to, the Piccolo of Abaxis company, the Afinion of Axis-Shield company etc., this type of POCT analyser can realize that all but small amount of sample can be analyzed, easy and simple to handle, no crossed contamination automated job, is highly suitable for China and has huge primary care system and reagent usage quantity country greatly like this.
Along with Eleventh Five-Year Plan plan purchasing and strengthen at three grades and second-grade hospital infrastructure device, middle rank possesses good software and hardware facilities to go to the hospital at present, but basic medical unit particularly its full-automatic biochemical testing instruments facility of unit such as commune hospital, clinic is generally not enough, the check professional who does not also have enough numbers, therefore setting up operation POCT analytical system simple and easy, cheap, real-time report has important meaning to bringing into play the effect of vast basic hospital in disease prevention and diagnosis and treatment.And provide a kind of bilirubin direct detection reagent that can be used for the POCT analyser of above-mentioned introducing micro-fluidic chip technology to become problem demanding prompt solution.
Summary of the invention
The present invention is directed to the above-mentioned deficiency of prior art, provide a kind of DBIL that can be used for introducing the POCT analyser of micro-fluidic chip technology to measure reagent.
The present invention can be used for the DBIL mensuration reagent of above-mentioned POCT analyser (introducing the micro-fluidic chip technology) and realizes by following technical scheme: a kind of bilirubin direct detection reagent of the POCT of can be used for analyser, comprise diluent and reaction reagent, wherein diluent is to be grouped into by following one-tenth:
Damping fluid 1(pH6.5-7.5) 0.01-1.0mol/L,
Tensio-active agent 0.1-10.0%(mass percent),
Sanitas 0.1-10.0%(mass percent),
Ascorbic acid oxidase 1-100KU/L;
Wherein said reaction reagent is to be grouped into by following one-tenth:
Damping fluid 2(pH3.0-5.0) 0.1-1.0mol/L,
Sodium-chlor 45g/L,
Disodium ethylene diamine tetraacetate 2-10g/L,
Oxammonium sulfate 2-10g/L,
Etidronic acid 2-10g/L,
Sodium Persulfate 100-200g/L,
Sulfuric acid 0.1-2ml/L,
Sanitas 0.1-1g/L,
Lyophilized vaccine 10-30g/L.
Wherein said damping fluid 1 can be the amino damping fluid of trishydroxymethyl, glycine-NaOH damping fluid, N-2-hydroxyethyl piperazine-N '-2-ethanesulfonic acid damping fluid, N-three (methylol) methylamino--2-hydroxy-propanesulfonic acid damping fluid, N-three (methylol) methyl-2-aminoethyl sulfonic acid damping fluid, piperazine-N, two (2-hydroxyethanesulfonic acid) damping fluids of N-, 3-morpholine-2-hydroxypropionate sodium damping fluid, 3-(N-morpholine) ethyl sulfonic acid sodium damping fluid, the 4-(2-hydroxyethyl) piperazine-1-2-hydroxy-propanesulfonic acid damping fluid, N-(2-hydroxyethyl) piperazine-N '-4-fourth sulfonic acid damping fluid, two (2-hydroxyethyl) amino of 3--2-hydroxy-propanesulfonic acid damping fluid, 3-(encircling amine)-2-hydroxyl-1-propanesulfonic acid damping fluid, 4-(2-hydroxyethyl)-1-piperazine propanesulfonic acid damping fluid, 3-(encircling amine)-1-propanesulfonic acid damping fluid, 3-morpholine propanesulfonic acid damping fluid, N-three (methylol) methyl-3-aminopropanesulfonicacid acid damping fluid a kind of.
Wherein said damping fluid 2 can be a kind of in 2-(N-morpholine) ethyl sulfonic acid damping fluid, acetate buffer, sodium citrate buffer solution, the phosphoric acid buffer.
Described tensio-active agent is nonionogenic tenside, as be selected from TWEEN series (as tween (TWEEN) 20, polysorbate40, polysorbate60, tween 80), SPAN series is (as (SPAN) 20 of class of department, class of department 40, class of department 60, class of department 80), TRITON series is (as Triton X-100, Triton X-114, Triton X-405) one or more of concrete material (namely can be TWEEN series in, a kind of concrete material in SPAN series or the TRITON series, or more than one concrete material in every kind of series, or a kind of concrete material in two kinds or the above series).
Described sanitas (comprising sanitas in the diluent and the sanitas in the reaction reagent) is selected from potassium sorbate, Sodium Benzoate, Sodium Nitrite, the proclin series sanitas (as Proclin300) a kind of in a kind of or nipagin esters (as methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propylparaben, butyl p-hydroxybenzoate, p-Hydroxybenzoic acid isopropyl ester, p-Hydroxybenzoic acid isobutyl ester).
Described lyophilized vaccine selects one or several in trehalose, sucrose, bovine serum albumin, tween 80, triton x-100, the fatty alcohol-polyoxyethylene ether (Brij-35).
Diluent is liquid state in the described DBIL mensuration reagent, and reaction reagent is dry powder.
The preparation method of the diluent of described DBIL mensuration reagent is as follows: stir evenly mixing behind the described reagent composition adding distilled water.
The preparation method of reaction reagent that described DBIL measures reagent is as follows: described reagent composition is added to mix behind the distilled water stir evenly, 0.5-10 μ l reaction reagent is joined the reaction detection groove, volatilize 24-72h through freeze-drying (industry routine techniques) or 2-8 ℃.
The test condition that described DBIL measures reagent is as follows: temperature: 37 ℃; Detect predominant wavelength 450nm, commplementary wave length 750nm.
Micro-fluidic chip technology of the present invention is that basic operation unit is integrated on the chip of one tens square centimeters (even littler), forms network by the microchannel, runs through a kind of technology of total system with controlled fluid.It is two-layer up and down that its characteristics are that chip generally is divided into, the upper strata is useful on the through hole of application of sample, and lower floor comprises the respond difform fluid channel etc. of the reaction detection groove of reagent, one group of self check groove that is used for system compensation, one group of overflow groove, many group control fluid flow of sample cell, dilution liquid bath, sample quantitative slot, diluent quantitative slot, reservoir, a plurality of prepackage.Its detection method generally comprises following steps: (1) is injected into sample solution and diluent in described sample cell and the dilution liquid bath through through hole separately; (2) starter motor rotates described chip; (3) sample solution is realized solid-liquid separation and quantitative under centrifugal action, and diluent enters the diluent quantitative slot simultaneously; (4) quantified sample is mixed with diluent inflow tempering tank; (5) mixed liquid enters the reaction detection groove and reaction reagent reacts; (6) by in the reaction detection groove, carrying out in situ detection with the supporting test set of chip.
The measuring method of described DBIL mensuration reagent is as follows: 5-20 μ l sample joined sample cell, 20-100 μ l diluent joined the dilution liquid bath, and starter motor, record absorbance A 1 behind 37 ℃ of reaction 1min continues to record absorbance A 2 behind the reaction 5-9min.
The reaction principle that described DBIL measures reagent is: sample mixes with diluent earlier hatches, with interfering substances such as the vitamins C in the removal sample and blood fat, after mixed solution enters the reaction detection groove, in the presence of sodium-chlor, disodium ethylene diamine tetraacetate, oxammonium sulfate, etidronic acid and sulfuric acid, oxidized dose of Sodium Persulfate of the bilirubin direct in the sample is oxidized to uteroverdine.Bilirubinic yellow specificity absorbancy descends, and by measuring the variation of Sodium Persulfate oxidation front and back absorbancy, calculates the content of DBIL in the sample.
Advantage of the present invention and beneficial effect: reagent of the present invention has good sensitivity, accuracy, precision and linearity, can satisfy the Clinical Laboratory requirement fully.Reagent of the present invention can be used for introducing the POCT analyser of micro-fluidic chip technology, thereby realizes that operation is simple and easy, cheap, the foundation of the POCT analytical system of real-time report.
Description of drawings
Fig. 1 detects with the analyte that the high density serum specimen of collecting from hospital is diluted to different concns, the linear figure as a result of gained DBIL.
The DBIL end value of measuring on Fig. 2 and the automatic clinical chemistry analyzer compares the figure as a result of gained, and wherein X-axis represents determination data in the automatic clinical chemistry analyzer Hitachi 7180, and Y-axis represents determination data on the POCT analyser.
Embodiment
To further specify the present invention by following non-limiting example below, as well known to those skilled in the art, without departing from the spirit of the invention, can make many modifications to the present invention, such modification also falls into scope of the present invention.
Following experimental technique is ordinary method if no special instructions, and employed experiment material all can easily be obtained from commercial company if no special instructions.
Diluent:
Amino damping fluid (pH7.0) 0.1mol/L of trishydroxymethyl
TWEEN80 5%
Proclin300 0.1%
Ascorbic acid oxidase 1KU/L
Reaction reagent:
Phosphoric acid buffer (pH4.0) 0.1mol/L
Sodium-chlor 45g/L
Disodium ethylene diamine tetraacetate 2g/L
Oxammonium sulfate 2g/L
Etidronic acid 2g/L
Sodium Persulfate 100g/L
Sulfuric acid 0.1ml/L
Potassium sorbate 0.1g/L
Bovine serum albumin 5g/L.
Diluent:
Amino damping fluid (pH7.5) 0.5mol/L of trishydroxymethyl
TRITONX-100 5%
Methyl p-hydroxybenzoate 1%
Ascorbic acid oxidase 2KU/L
Reaction reagent:
2-(N-morpholine) ethyl sulfonic acid sodium damping fluid (pH3.0) 0.5mol/L
Sodium-chlor 45g/L
Disodium ethylene diamine tetraacetate 10g/L
Oxammonium sulfate 5g/L
Etidronic acid 5g/L
Sodium Persulfate 150g/L
Sulfuric acid 1ml/L
Proclin300 1g/L
Trehalose 10g/L.
Embodiment 3
Diluent:
3-morpholine propanesulfonic acid damping fluid (pH6.5) 1.0mol/L
SPAN20 10%
Ascorbic acid oxidase 10KU/L
Reaction reagent:
Sodium citrate buffer solution (pH5.0) 1mol/L
Sodium-chlor 45g/L
Disodium ethylene diamine tetraacetate 5g/L
Oxammonium sulfate 10g/L
Etidronic acid 10g/L
Sodium Persulfate 50g/L
Sulfuric acid 1ml/L
Proclin300 1g/L
Fatty alcohol-polyoxyethylene ether 10g/L.
Describe below in conjunction with the performance of form to the embodiment of the invention 1 gained reagent.
1, precision
Table 1, precision assessment result
2, linearity
The analyte that is diluted to different concns with the high density serum specimen of collecting from hospital detects, and gained DBIL linearity the results are shown in Fig. 1.
3, methodology comparison test
Compare with the DBIL end value of measuring on the automatic clinical chemistry analyzer, result such as Fig. 2, wherein X-axis represents determination data in the automatic clinical chemistry analyzer Hitachi 7180, POCT analyser (the living world of Taiwan guarantor of micro-fluidic chip technology is introduced in the Y-axis representative, Amishield TMO-100) goes up determination data.
4, detection sensitivity
The appraisal procedure of the detection sensitivity standard deviation of 10-20 dummy signal strength, and the standard deviation of 3-15 least significant non-zero sample signal strength are calculated gained with the EP Evaluator release6 of statistical software.Experimental result shows that reagent of the present invention has good sensitivity, can meet the improvement of U.S. clinical labororatory fully and amend legislation.
Table 2 reagent detection sensitivity
From above-mentioned detected result as can be known, reagent of the present invention has good sensitivity, accuracy, precision and linearity, can satisfy the Clinical Laboratory requirement fully.
Claims (9)
1. bilirubin direct detection reagent, it is characterized in that: this reagent comprises diluent and reaction reagent, wherein diluent is to be grouped into by following one-tenth:
Damping fluid 1(pH6.5-7.5) 0.01-1.0mol/L,
Tensio-active agent 0.1-10.0%(mass percent),
Sanitas 0.1-10.0%(mass percent),
Ascorbic acid oxidase 1-100KU/L;
Wherein said reaction reagent is to be grouped into by following one-tenth:
Damping fluid 2(pH3.0-5.0) 0.1-1.0mol/L,
Sodium-chlor 45g/L,
Disodium ethylene diamine tetraacetate 2-10g/L,
Oxammonium sulfate 2-10g/L,
Etidronic acid 2-10g/L,
Sodium Persulfate 100-200g/L,
Sulfuric acid 0.1-2ml/L,
Sanitas 0.1-1g/L,
Lyophilized vaccine 10-30g/L.
2. bilirubin direct detection reagent according to claim 1, it is characterized in that: described damping fluid 1 is the amino damping fluid of trishydroxymethyl, glycine-NaOH damping fluid, N-2-hydroxyethyl piperazine-N '-2-ethanesulfonic acid damping fluid, N-three (methylol) methylamino--2-hydroxy-propanesulfonic acid damping fluid, N-three (methylol) methyl-2-aminoethyl sulfonic acid damping fluid, piperazine-N, two (2-hydroxyethanesulfonic acid) damping fluids of N-, 3-morpholine-2-hydroxypropionate sodium damping fluid, 3-(N-morpholine) ethyl sulfonic acid sodium damping fluid, the 4-(2-hydroxyethyl) piperazine-1-2-hydroxy-propanesulfonic acid damping fluid, N-(2-hydroxyethyl) piperazine-N '-4-fourth sulfonic acid damping fluid, two (2-hydroxyethyl) amino of 3--2-hydroxy-propanesulfonic acid damping fluid, 3-(encircling amine)-2-hydroxyl-1-propanesulfonic acid damping fluid, 4-(2-hydroxyethyl)-1-piperazine propanesulfonic acid damping fluid, 3-(encircling amine)-1-propanesulfonic acid damping fluid, 3-morpholine propanesulfonic acid damping fluid, a kind of in N-three (methylol) the methyl-3-aminopropanesulfonicacid acid damping fluid.
3. bilirubin direct detection reagent according to claim 1 is characterized in that: a kind of in 2-(N-morpholine) ethyl sulfonic acid damping fluid, acetate buffer, sodium citrate buffer solution, the phosphoric acid buffer of described described damping fluid 2.
4. bilirubin direct detection reagent according to claim 1 is characterized in that: described tensio-active agent is to be selected from one or more of concrete material in TWEEN series, SPAN series, the TRITON series.
5. bilirubin direct detection reagent according to claim 1 is characterized in that: described sanitas is a kind of in a kind of or nipagin esters in potassium sorbate, Sodium Benzoate, Sodium Nitrite, the proclin series sanitas.
6. bilirubin direct detection reagent according to claim 5 is characterized in that: described proclin series sanitas is Proclin300.
7. bilirubin direct detection reagent according to claim 5 is characterized in that: described nipagin esters is a kind of in methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propylparaben, butyl p-hydroxybenzoate, p-Hydroxybenzoic acid isopropyl ester, the p-Hydroxybenzoic acid isobutyl ester.
8. bilirubin direct detection reagent according to claim 1, it is characterized in that: described lyophilized vaccine is one or several in trehalose, sucrose, bovine serum albumin, tween 80, triton x-100, the fatty alcohol-polyoxyethylene ether.
9. bilirubin direct detection reagent according to claim 1, it is characterized in that: the diluent in the described bilirubin direct detection reagent is liquid state, reaction reagent is dry powder.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104406971A (en) * | 2014-11-28 | 2015-03-11 | 山东博科生物产业有限公司 | Direct bilirubin detection reagent |
| CN104777116A (en) * | 2015-04-14 | 2015-07-15 | 山东博科生物产业有限公司 | Total bilirubin detection kit with strong anti-interference performance |
| CN106404686A (en) * | 2016-08-27 | 2017-02-15 | 山东博科生物产业有限公司 | Antiheparin serum total bilirubin (vanadate oxidation method) detection 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 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104406971A (en) * | 2014-11-28 | 2015-03-11 | 山东博科生物产业有限公司 | Direct bilirubin detection reagent |
| CN104406971B (en) * | 2014-11-28 | 2017-03-08 | 山东博科生物产业有限公司 | A kind of bilirubin direct detectable |
| CN104777116A (en) * | 2015-04-14 | 2015-07-15 | 山东博科生物产业有限公司 | Total bilirubin detection kit with strong anti-interference performance |
| CN106404686A (en) * | 2016-08-27 | 2017-02-15 | 山东博科生物产业有限公司 | Antiheparin serum total bilirubin (vanadate oxidation method) detection kit |
| CN106404686B (en) * | 2016-08-27 | 2019-03-29 | 山东博科生物产业有限公司 | A kind of serum total bilirubin of antiheparin (Vanadic acid oxidation) detection 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 |
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