CN104048929A - Detection kit for total bilirubin - Google Patents
Detection kit for total bilirubin Download PDFInfo
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- CN104048929A CN104048929A CN201310075760.4A CN201310075760A CN104048929A CN 104048929 A CN104048929 A CN 104048929A CN 201310075760 A CN201310075760 A CN 201310075760A CN 104048929 A CN104048929 A CN 104048929A
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- total bilirubin
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Abstract
The invention discloses a detection kit for total bilirubin through a comprehensive oxidizing agent method. The kit is a liquid-type dual reagent which is composed of a reagent 1 and a reagent 2 with a ratio being 4:1, wherein the reagent 1 comprises 10-100 mmol/L of a Tris-HCl buffer solution and 0.1-5 mmol/L, and the reagent 2 comprises 5-40 mmol/L of a phosphatic buffer solution and 0.2-6 mmol/L of a chemical oxidizing agent. The detection kit for total bilirubin has advantages of a high oxidizing reaction speed, a large signal, an accurate result and a strong anti-interference capability.
Description
Technical field
The present invention relates to a kind of kit of measuring total bilirubin in serum, is specifically the kit that a kind of complex oxidant method is measured total bilirubin.
Background technology
Total bilirubin is combination and the bilirubinic general name of non-binding type being produced by protoheme metabolism in human body, and whether whether be mainly used to diagnosis has liver diseases or biliary tract to occur extremely, and the reason that total bilirubin is on the low side is likely because hypoferric anemia.Therefore, the mensuration to total bilirubin in the serum of human body, blood plasma and urine, is one of the most conventional project of medical test.
Total bilirubin determination is an important test item in liver, courage functional check.The method of measuring at present serum bilirubin mainly contains diazonium method, enzyme process.Diazonium method reagent is unstable, must be prepared by sodium nitrite and sulfaminic acid, and the holding time is shorter temporarily; The 2nd, diazo reagent is not good with the specificity that indirect bilirubin and bilirubin direct (DBIL) react, and while measuring DBIL under certain condition, has a small amount of indirect bilirubin to participate in reaction, if the reaction time extends, participation more, cause measured value inaccurate.Enzyme process price is more expensive, and active decline also easily occurs for bilirubin oxidase wherein, the accuracy that impact is measured.
Chemical oxidization method has vanadate oxidizing process and nitrite-oxidizing method.Vanadic acid oxidizing process has the features such as stable reagent, stable reaction, anti-interference (Hb) ability are strong, but Vanadium hydrochlorate method is subject to the isoionic interference of ALT AST, and Vanadium ion is heavy metal, easily contaminated environment.Nitrite-oxidizing method more can reduce the interference producing because of oxidation reaction compared with vanadic acid oxidizing process.But vanadate method and nitrite method are subject to a courage, and false positive, easily appears in interference as unconjugated bilirubin, δ-cholerythrin etc. clinically.
Summary of the invention
The object of the invention is to overcome the defect that prior art exists, provide a kinds of oxidation reaction speed fast, signal is large, and result is accurate, the total bilirubin detection kit that antijamming capability is stronger.
The technical scheme that realizes the object of the invention is: a kind of total bilirubin detection kit is provided, and the liquid double reagent that this kit is comprised of reagent 1 and reagent 2, wherein each component of reagent 1 and concentration range are:
Tris-HCl damping fluid 10 ~ 100mmol/L
Surfactant 0.1 ~ 5mmmol/L
Each component and the concentration range thereof of reagent 2 are:
Phosphate buffer 5 ~ 40mmol/L
Chemical oxidizing agent 0.2 ~ 6mmol/L
As preferably, total bilirubin detection kit of the present invention, wherein the each component of reagent 1 and concentration range are:
Tris-HCl damping fluid 30 ~ 80mmol/L
Surfactant 0.8-3.1g/L
Wherein each component of reagent 2 and concentration range thereof are:
Phosphate buffer 12 ~ 35mmol/L
Chemical oxidizing agent 0.8 ~ 3.2mmol/L
As most preferably, total bilirubin detection kit of the present invention, wherein each component of reagent 1 and concentration range are:
Tris-HCl damping fluid 50mmol/L
Surfactant 2mmol/L
Wherein each component of reagent 2 and concentration range thereof are:
Phosphate buffer 1 8mmol/L
Chemical oxidizing agent 2mmol/L
The detection kit of total bilirubin of the present invention, the volume ratio of described reagent 1 and reagent 2 is: 4:1.
The detection kit reagent 1 of total bilirubin of the present invention and the raw material in reagent 2 are selected according to being:
In above component, Tris-HCl damping fluid and phosphate buffer are the damping fluid of reagent 1 and reagent 2, are beneficial to the dispersion of various compositions in reagent, and it improves the stability of reagent.Surfactant, is the promoter of reaction, and the impact that can remove other lipid impurity, improves the sensitivity detecting.Chemical oxidizing agent, is that multiple oxygenant combines, and has added corresponding composition, it makes the electronic match ability of oxidation reaction stronger, and sensitivity is higher.
The collocation method of kit reagent 1 of the present invention and reagent 2 is conventional method, and reagent 1 and reagent 2 mix and stir evenly separately after adding respectively distilled water.
With kit of the present invention, by following method, measure total bilirubin, total bilirubin, under surfactant exists, is oxidized by chemical oxidizing agent, generates dehydrobilirubin, measures thus the reduction of 450nm place absorbance, in the hope of the concentration of total bilirubin in sample.
The assay method of applying total bilirubin in total bilirubin detection kit mensuration sample of the present invention is as follows: sample reagent adding 1 mixes, and after 30 ~ 40 ℃ of reaction 3 ~ 5min, reads absorbance A
1, add reagent 2 to mix, after 30 ~ 40 ℃ of reaction 3 ~ 5min, read absorbance A
2, Δ A=A
2-A
1.Amount of samples 8 μ L wherein, reagent 1 consumption 200-300 μ L, reagent 2 consumption 30-110 μ L.
The cubage formula of applying total bilirubin in total bilirubin detection kit mensuration sample of the present invention is as follows:
Total bilirubin=sample absorbance Δ A * concentration of standard solution/titer absorbance Δ A
0
The present invention has positive effect: compare with existing total bilirubin detection kit, specificity of the present invention is better, and antijamming capability is stronger, and linearity is advantages of higher more, is applicable to automatic clinical chemistry analyzer, has larger clinical value.
Accompanying drawing explanation
For content of the present invention is more easily expressly understood, according to specific embodiment also by reference to the accompanying drawings, the present invention is further detailed explanation, wherein below
Fig. 1 is the real time reaction curve map that total bilirubin detection kit of the present invention is measured total bilirubin.
Fig. 2 is the real time reaction curve map that contrast agent box is measured total bilirubin.
In Fig. 1, Fig. 2, horizontal ordinate is the moment of Hitachi's 7060 automatic clinical chemistry analyzer reaction monitorings, by 10 minutes, is divided into 31 points, and each point has the time point of absorbance reaction, and ordinate is absorbance A.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail, but the present invention is far not limited only to following examples.
Embodiment 1
Reagent 1(R1) each component and concentration range thereof:
R1 Tris-HCl damping fluid 10mmol/L
Surfactant 0.1mmol/L
Reagent 2(R2) each component and concentration range thereof are as follows:
R2 phosphate buffer 5mmol/L
Chemical oxidizing agent 0.2mmol/L
The collocation method of reagent 1 and reagent 2 is conventional method, and the component described in reagent 1 and reagent 2 is mixed and stirred evenly separately after adding respectively distilled water.
Embodiment 2
Reagent 1(R1) each component and concentration range thereof:
R1 Tris-HCl damping fluid 100mmol/L
Surfactant 5mmol/L
Reagent 2(R2) each component and concentration range thereof are as follows:
R2 phosphate buffer 40mmol/L
Chemical oxidizing agent 6mmol/L
The preparation method of embodiment 2 kits is with embodiment 1.
Embodiment 3
Reagent 1(R1) each component and concentration range thereof:
R1 Tris-HCl damping fluid 30mmol/L
Surfactant 0.8mmol/L
Reagent 2(R2) each component and concentration range thereof are as follows:
R2 phosphate buffer 1 2mmol/L
Chemical oxidizing agent 0.8mmol/L
The preparation method of embodiment 3 kits is with embodiment 1.
Embodiment 4
Reagent 1(R1) each component and concentration range thereof:
R1 Tris-HCl damping fluid 80mmol/L
Surfactant 3.1mmol/L
Reagent 2(R2) each component and concentration range thereof are as follows:
R2 phosphate buffer 1 2mmol/L
Chemical oxidizing agent 3.2mmol/L
The preparation method of embodiment 4 kits is with embodiment 1.
Embodiment 5
Reagent 1(R1) each component and concentration range thereof:
R1 Tris-HCl damping fluid 50mmol/L
Surfactant 2mmol/L
Reagent 2(R2 wherein) each component and concentration range thereof:
R2 phosphate buffer 1 8mmol/L
Chemical oxidizing agent 2mmol/L
The preparation method of embodiment 5 kits is with embodiment 1.
The assay method of applying total bilirubin in total bilirubin detection kit mensuration sample of the present invention is as follows: sample reagent adding 1 mixes, and after 30 ~ 40 ℃ of reaction 3 ~ 5min, reads absorbance A
1, add reagent 2 to mix, after 30 ~ 40 ℃ of reaction 3 ~ 5min, read absorbance A
2, Δ A=A
2-A
1.Wherein, amount of samples 8 μ L, reagent 1 consumption 200-300 μ L, reagent 2 consumption 30-110 μ L.
Finally, computation formula:
Formula total bilirubin=sample absorbance Δ A * concentration of standard solution/titer absorbance Δ A
0
Obtain total bilirubin detection kit of the present invention and measure the content of total bilirubin in sample.
Below in conjunction with Fig. 1, Fig. 2 and form, the performance of the kit of comparative example and kit of the present invention is compared.These two kinds of kits are compared in impact by response curve, sensitivity, the range of linearity, accuracy and various chaff interferences etc.
Its composition of comparative example kit and formula are:
Reagent 1:Good's acid buffer 150mmol/L
Reaction promoter, anti-interference agent
Reagent 2: oxygenant 10.0mmol/L
Non-specific responding inhibitor 30.0mmol/L
Kit of the present invention: reagent 1 and reagent 2.
The comparison of the range of linearity of two kinds of kits:
Preparation 0,50,100,150,200,250,300,350,400 μ mol/L total bilirubin standard solution, measure its concentration by total bilirubin detection kit of the present invention and contrast agent box respectively, and result is as follows:
Table 1 kit of the present invention and contrast agent box are measured range of linearity contrast
The kit that the range of linearity of kit of the present invention contrasts is high by 100.0%.When total bilirubin concentration is greater than 250 μ mol/L, contrast agent box can not detect, and the range of linearity is too small, illustrates that kit of the present invention has linear test specification large.
2, the comparison of the sensitivity of two kinds of kits:
With the quality-control product of existing kit and kit measurement of the present invention 75.2 μ mol/L, poor (A) reading of absorbance is as follows:
| Absorbance | A 1 | A 2 | ?A=A 2-A 1 |
| Kit of the present invention | -0 | -1480 | -1480 |
| Contrast agent box | -0.01 | -1150 | -1149.99 |
As can be seen from the above table, when kit of the present invention is measured same sample compared with contrast agent box, absorbance is higher, shows that the present invention has higher sensitivity.
3, the accuracy comparison of two kinds of kits:
With the calibration of Roche calibration object, with kit of the present invention and contrast agent box, measure the high and low value Quality Control of Roche respectively.Result is as follows:
Table 2 kit of the present invention and the contrast of contrast agent box measurement result
The accuracy of measurement of the kit of kit of the present invention and contrast is all in 2SD, and kit accuracy of the present invention is higher.
4, the comparison of the anti-interference of two kinds of kits
In definite value pooled serum (13.2 μ mol/L), add haemoglobin, vitamin C, Intralipos, four kinds of chaff interferences of triglyceride, measure respectively with kit of the present invention and contrast agent box, measurement result is as follows:
Table 3 kit of the present invention and contrast agent box are measured the contrast of antijamming capability result
To contrast total bilirubin detection kit antijamming capability stronger for kit of the present invention as can be seen from the above table.
From the known kit of the present invention of above-described embodiment, have oxidation reaction speed fast, signal is large, and specificity is better, and antijamming capability is strong, highly sensitive, the advantage that the range of linearity is large.
Above-described specific embodiment; object of the present invention, technical scheme and beneficial effect are further described; institute is understood that; the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any modification of making, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (4)
1. a total bilirubin detection kit, is characterized in that: the liquid-type double reagent that this kit is comprised of reagent 1 and reagent 2, and wherein each component of reagent 1 and concentration range are:
Tris-HCl damping fluid 10 ~ 100mmol/L
Surfactant 0.1 ~ 5mmmol/L
Each component and the concentration range thereof of reagent 2 are:
Phosphate buffer 5 ~ 40mmol/L
Chemical oxidizing agent 0.2 ~ 6mmol/L
2. total bilirubin detection kit according to claim 1, is characterized in that: each component and the concentration range of described reagent 1 are:
Tris-HCl damping fluid 30 ~ 80mmol/L
Surfactant 0.8-3.1g/L
Each component and the concentration range thereof of described reagent 2 are:
Phosphate buffer 12 ~ 35mmol/L
Chemical oxidizing agent 0.8 ~ 3.2mmol/L
3. total bilirubin detection kit according to claim 2, is characterized in that: each component and the concentration range of described reagent 1 are:
Tris-HCl damping fluid 50mmol/L
Surfactant 2mmol/L
Each component and the concentration range thereof of described reagent 2 are:
Phosphate buffer 1 8mmol/L
Chemical oxidizing agent 2mmol/L
4. according to the total bilirubin detection kit described in claim 1,2 or 3, it is characterized in that: the volume ratio of described reagent 1 and reagent 2 is: 4:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310075760.4A CN104048929A (en) | 2013-03-11 | 2013-03-11 | Detection kit for total bilirubin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310075760.4A CN104048929A (en) | 2013-03-11 | 2013-03-11 | Detection kit for total bilirubin |
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|---|---|
| CN104048929A true CN104048929A (en) | 2014-09-17 |
Family
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|---|---|---|---|
| CN201310075760.4A Pending CN104048929A (en) | 2013-03-11 | 2013-03-11 | Detection kit for total bilirubin |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN105092573A (en) * | 2015-09-02 | 2015-11-25 | 郁东 | Novel direct bilirubin detection kit |
| CN105586384A (en) * | 2016-03-11 | 2016-05-18 | 威尚生物技术（合肥）有限公司 | Total bilirubin detection kit |
| 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 |
| CN106896101A (en) * | 2015-12-21 | 2017-06-27 | 徐淼 | A kind of TBA detection reagent |
| CN111424070A (en) * | 2020-03-03 | 2020-07-17 | 天津大学 | Total bilirubin detection kit containing bacillus subtilis laccase |
| CN114277088A (en) * | 2021-12-02 | 2022-04-05 | 深圳市锦瑞生物科技股份有限公司 | Total bilirubin determination reagent, preparation method of reagent ball and determination chip |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0087150A1 (en) * | 1982-02-22 | 1983-08-31 | Seragen Diagnostics, Inc. | Process, reagent and kit for the assay of total bilirubin |
| CN1959415A (en) * | 2006-10-30 | 2007-05-09 | 宁波美康生物科技有限公司 | Kit for mensurating total bilirubin through chemistry oxidation process |
| CN101144824A (en) * | 2007-10-10 | 2008-03-19 | 宁波美康生物科技有限公司 | Total bilirubin determination reagent kit |
| CN101776692A (en) * | 2009-01-09 | 2010-07-14 | 上海复星医药（集团）股份有限公司 | Kit for detecting total bilirubin by using vanadate oxidation method |
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2013
- 2013-03-11 CN CN201310075760.4A patent/CN104048929A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| EP0087150A1 (en) * | 1982-02-22 | 1983-08-31 | Seragen Diagnostics, Inc. | Process, reagent and kit for the assay of total bilirubin |
| CN1959415A (en) * | 2006-10-30 | 2007-05-09 | 宁波美康生物科技有限公司 | Kit for mensurating total bilirubin through chemistry oxidation process |
| CN101144824A (en) * | 2007-10-10 | 2008-03-19 | 宁波美康生物科技有限公司 | Total bilirubin determination reagent kit |
| CN101776692A (en) * | 2009-01-09 | 2010-07-14 | 上海复星医药（集团）股份有限公司 | Kit for detecting total bilirubin by using vanadate oxidation method |
Non-Patent Citations (2)
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| 白峰等: "胆红素氧化酶法测定血清总胆红素试剂盒在Monarch自动生化分析仪上的应用评价", 《陕西医学检验》 * |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN105092573A (en) * | 2015-09-02 | 2015-11-25 | 郁东 | Novel direct bilirubin detection kit |
| CN106896101A (en) * | 2015-12-21 | 2017-06-27 | 徐淼 | A kind of TBA detection reagent |
| CN105586384A (en) * | 2016-03-11 | 2016-05-18 | 威尚生物技术（合肥）有限公司 | Total bilirubin detection kit |
| 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 |
| CN111424070A (en) * | 2020-03-03 | 2020-07-17 | 天津大学 | Total bilirubin detection kit containing bacillus subtilis laccase |
| CN114277088A (en) * | 2021-12-02 | 2022-04-05 | 深圳市锦瑞生物科技股份有限公司 | Total bilirubin determination reagent, preparation method of reagent ball and determination chip |
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