CN107741491A - A kind of kit for determining urine galactose and its preparation application method - Google Patents
A kind of kit for determining urine galactose and its preparation application method Download PDFInfo
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- CN107741491A CN107741491A CN201710917819.8A CN201710917819A CN107741491A CN 107741491 A CN107741491 A CN 107741491A CN 201710917819 A CN201710917819 A CN 201710917819A CN 107741491 A CN107741491 A CN 107741491A
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The invention discloses a kind of kit for determining urine galactose, including reagent R1 independent of each other and reagent R2 biliquid components, including composition and corresponding content be：Reagent R1：KH2PO44 6g/L, DETA.2Na0.5 1.0g/L, BSA0.5 1.0g/L, the 3.0g/L of P-hydroxybenzoic acid sodium 2.0, solvent is purified water；Reagent R2：MOPSO 5 8g/L, MOPSO.Na 6 8g/L, POD 20 80U L, the 1.0g/L of 4 ATP 0.3, BSA0.2 0.8g/L, the 400U of GAD 100, solvent is purified water.The invention also discloses a kind of preparation application method for the kit for determining urine galactose.Advantage of the present invention is：Detection speed is fast, efficiency high, suitable for automatic clinical chemistry analyzer.
Description
Technical field
The present invention relates to medical science and technological field of biochemistry, more particularly to a kind of survey suitable for full-automatic biochemical analysis
Determine the kit of urine galactose and its prepare application method.
Background technology
Lactose intolerance is due to lactose digestion and malabsorption caused by the shortage lactase of small intestinal mucosa surface, and with
A series of clinical symptoms such as abdominal distension, diarrhoea, stomachache.The reason for according to occurring, lactose intolerance can be divided into：Congenital lactase
Shortage, adult type alactasia and secondary lactase deficiency.Wherein adult type alactasia is the master of lactose intolerance
Type is wanted, it may is that eating habit causes the expression of lactase gene to be progressively closed off with the extension of time, performance
To be gradually reduced with the growth at age, lactase activity, until disappearing.
The test in laboratory method of lactose intolerance has many kinds, currently used to have hydrogen breath test, stool normal
Rule chemical examination, reducing sugar test and urine galactose experiment.Wherein, urine galactose experiment is by determining the galactose concentration in urine
Reflect the lactose tolerance digested and assimilated situation, judge subject of lactose indirectly.Using urine galactose level as index, human body is most
Suitable concentration is 41-116 μm of ol/24h.
Presently, urine galactose is determined mainly to carry out using traditional gold label test strip method.The method operation is more multiple
It is miscellaneous, and human factor has a great influence to experimental result, can not be applied to automatic clinical chemistry analyzer, needs manual operations, test is delayed
Slowly, the qualification of operating personnel has a great influence to result, can not carry out and promote on a large scale.
Therefore, it is badly in need of that a kind of detection speed is fast, efficiency high at present, urinates gala suitable for the measure of automatic clinical chemistry analyzer
The kit and its preparation application method of sugar.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of detection speed is fast, efficiency high, it is applied to
The kit and its preparation application method of the measure urine galactose of automatic clinical chemistry analyzer.
The present invention is achieved by the following technical solutions：A kind of kit for determining urine galactose, including it is independent of one another
Reagent R1 and reagent R2 biliquid components, including composition and corresponding content be：
Reagent R1：
Reagent R2：
One of preferred embodiment as the present invention, including reagent R1 independent of each other and reagent R2 biliquid components, including
Composition and corresponding content be：
Reagent R1：
Reagent R2：
One of preferred embodiment as the present invention, including reagent R1 independent of each other and reagent R2 biliquid components, including
Composition and corresponding content be：
Reagent R1：
Reagent R2：
A kind of preparation method for the kit for determining urine galactose, comprises the following steps：
(1) reagent preparation R1
A. according to reagent R1 constituent content, by KH2PO4It is dissolved in purified water, after stirring, is configured to R1 buffer solutions；
B. according to reagent R1 constituent content, BSA is slowly dissolved in obtained R1 buffer solutions, after stirring, then
DETA.2Na and P-hydroxybenzoic acid sodium are dissolved in wherein, that is, reagent R1 is made；
(2) reagent preparation R2
A. according to reagent R2 constituent content, MOPSO and MOPSO.Na are dissolved in purified water, after stirring, prepared
Into R2 buffer solutions；
B. according to reagent R2 constituent content, BSA is slowly dissolved in obtained R2 buffer solutions, after stirring, then
POD, 4-ATP, GAD are dissolved in wherein, mixed liquor stirs, that is, reagent R2 is made.
A kind of application method for the kit for determining urine galactose, comprises the following steps：
(1) testing sample is mixed with reagent R1,37 DEG C of incubation 5min；
(2) mixed again with reagent R2, it is fully reacted；
(3) reacted absorbance difference is determined with automatic clinical chemistry analyzer；
(4) concentration of the urine galactose in sample is calculated according to absorbance change value.
One of preferred embodiment as the present invention, in the step (1) and step (2), reagent R1 and reagent R2 are according to body
Product ratio 4:1 mixing.
One of preferred embodiment as the present invention, in the step (3), surveyed at dominant wavelength 546nm, commplementary wave length 660nm
Determine absorbance A 1, absorbance A 2 is determined after 30s.
One of preferred embodiment as the present invention, in the step (4), absorbance change value is A1 and A2 changing value,
That is Δ A.
Cleaning Principle：Under acid condition, production of the galactolipin by galactolipin carbohydrate oxidase (GAD) hydrolysis generation in urine
Thing develops the color through peroxidase (POD), 4-AA (4-ATP) and P-hydroxybenzoic acid sodium, produces quinochrome, passes through
The increase of the pigment is determined, the concentration of urine galactose can be obtained.
In formula：ΔATThe sample cell absorbance compared with blank tube absorbance
ΔASThe calibration pipe absorbance compared with blank tube absorbance
CSUGA concentration in calibration solution.
The present invention compared with prior art the advantages of be：
(1) urine sample of prior art needs digestion process more, and causing laboratory, the smell is awful；The urine of the present invention
Digestion process need not be carried out, bad smell will not be not only produced, also simplifies experimental procedure；
(2) the detection reaction time of prior art is 20min, of the invention then only need 10min, when greatly have compressed reaction
Between；
(3) prior art is not used on automatic clinical chemistry analyzer, it is of the invention then can be applied to full-automatic biochemical analysis
On instrument, it is adapted to full-automatic testing, can carries out and promote on a large scale.
Embodiment
Embodiments of the invention are elaborated below, the present embodiment is carried out lower premised on technical solution of the present invention
Implement, give detailed embodiment and specific operating process, but protection scope of the present invention is not limited to following implementation
Example.
Embodiment 1
A kind of kit of measure urine galactose of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Reagent R2：
Embodiment 2
A kind of kit of measure urine galactose of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Reagent R2：
Embodiment 3
A kind of kit of measure urine galactose of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Reagent R2：
Embodiment 4
A kind of kit of measure urine galactose of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Reagent R2：
Embodiment 5
A kind of preparation method of the kit of measure urine galactose of the present embodiment, comprises the following steps：
(1) reagent preparation R1
A. according to the constituent content of reagent R1 in above-described embodiment, by KH2PO4It is dissolved in purified water, after stirring, matches somebody with somebody
R1 buffer solutions are made；
B. according to the constituent content of reagent R1 in above-described embodiment, BSA is slowly dissolved in obtained R1 buffer solutions, stirred
After mixing uniformly, then DETA.2Na and P-hydroxybenzoic acid sodium be dissolved in wherein, that is, reagent R1 is made；
(2) reagent preparation R2
A. according to the constituent content of reagent R2 in above-described embodiment, MOPSO and MOPSO.Na are dissolved in purified water, stirred
After uniformly, R2 buffer solutions are configured to；
B. according to the constituent content of reagent R2 in above-described embodiment, BSA is slowly dissolved in obtained R2 buffer solutions, stirred
After mixing uniformly, then POD, 4-ATP, GAD be dissolved in wherein, mixed liquor stirs, that is, reagent R2 is made.
Embodiment 6
A kind of application method of the kit of measure urine galactose of the present embodiment, comprises the following steps：
(1) set blank tube group and sample cell group, each group processing mode as follows：
Sample cell group：
A. 8uL testing samples are mixed with the reagent R1 that 200uL above-described embodiments obtain, 37 DEG C of incubation 5min；
B. mixed again with the reagent R2 that 50uL above-described embodiments obtain, fully reaction；
Blank tube group：Testing sample is replaced with distilled water, other are the same as sample cell group；
(2) automatic clinical chemistry analyzer is used, absorbance A 1 is determined at dominant wavelength 546nm, commplementary wave length 660nm, after 30s
Determine absorbance A 2, Δ A=A2-A1；
(3) concentration of the urine galactose in sample is calculated according to absorbance change value Δ A.
Its Cleaning Principle is：Under acid condition, production of the galactolipin by the hydrolysis generation of galactolipin carbohydrate oxidase in urine
Thing develops the color through peroxidase, 4-AA and P-hydroxybenzoic acid sodium, quinochrome is produced, by determining the pigment
Increase, the concentration of urine galactose can be obtained.
In formula：ΔATThe sample cell absorbance compared with blank tube absorbance
ΔASThe calibration pipe absorbance compared with blank tube absorbance
CSUGA concentration in calibration solution.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should be included in the scope of the protection.
Claims (8)
1. a kind of kit for determining urine galactose, it is characterised in that including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Reagent R2：
2. the kit of measure urine galactose according to claim 1, it is characterised in that including reagent R1 independent of each other
With reagent R2 biliquid components, including composition and corresponding content be：
Reagent R1：
Reagent R2：
3. the kit of measure urine galactose according to claim 1, it is characterised in that including reagent R1 independent of each other
With reagent R2 biliquid components, including composition and corresponding content be：
Reagent R1：
Reagent R2：
A kind of 4. method of the kit of the measure urine galactose prepared as described in claim any one of 1-3, it is characterised in that
Comprise the following steps：
(1) reagent preparation R1
A. according to reagent R1 constituent content, by KH2PO4It is dissolved in purified water, after stirring, is configured to R1 buffer solutions；
B. according to reagent R1 constituent content, BSA is slowly dissolved in obtained R1 buffer solutions, after stirring, then will
DETA.2Na and P-hydroxybenzoic acid sodium are dissolved in wherein, that is, reagent R1 is made；
(2) reagent preparation R2
A. according to reagent R2 constituent content, MOPSO and MOPSO.Na are dissolved in purified water, after stirring, are configured to R2
Buffer solution；
B. according to reagent R2 constituent content, BSA is slowly dissolved in obtained R2 buffer solutions, after stirring, then will
POD, 4-ATP, GAD are dissolved in wherein, and mixed liquor stirs, that is, reagent R2 is made.
A kind of 5. method of the kit of measure urine galactose using as described in claim any one of 1-3, it is characterised in that
Comprise the following steps：
(1) testing sample is mixed with reagent R1,37 DEG C of incubation 5min；
(2) mixed again with reagent R2, it is fully reacted；
(3) reacted absorbance difference is determined with automatic clinical chemistry analyzer；
(4) concentration of the urine galactose in sample is calculated according to absorbance change value.
6. the application method of the kit of measure urine galactose according to claim 5, it is characterised in that the step
(1) and in step (2), reagent R1 and reagent R2 are according to volume ratio 4:1 mixing.
7. the application method of the kit of measure urine galactose according to claim 5, it is characterised in that the step
(3) in, absorbance A 1 is determined at dominant wavelength 546nm, commplementary wave length 660nm, absorbance A 2 is determined after 30s.
8. the application method of the kit of measure urine galactose according to claim 7, it is characterised in that the step
(4) in, absorbance change value be A1 and A2 changing value, i.e. Δ A.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710917819.8A CN107741491A (en) | 2017-09-30 | 2017-09-30 | A kind of kit for determining urine galactose and its preparation application method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710917819.8A CN107741491A (en) | 2017-09-30 | 2017-09-30 | A kind of kit for determining urine galactose and its preparation application method |
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| CN107741491A true CN107741491A (en) | 2018-02-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201710917819.8A Pending CN107741491A (en) | 2017-09-30 | 2017-09-30 | A kind of kit for determining urine galactose and its preparation application method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109557088A (en) * | 2019-01-11 | 2019-04-02 | 四川沃文特生物技术有限公司 | A kind of reagent card for detecting excrement lactose content and the excrement lactose detection method based on this |
| CN109991222A (en) * | 2019-04-29 | 2019-07-09 | 徐詹程 | A kind of infant urine galactolipin instrument and its detection method |
| CN118243646A (en) * | 2024-04-24 | 2024-06-25 | 武汉简为医疗科技有限公司 | Lactose intolerance detection kit and application method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101324581A (en) * | 2007-06-13 | 2008-12-17 | 苏州艾杰生物科技有限公司 | Method for determining galactose concentration and galactose diagnosis/determination reagent kit |
| CN105400862A (en) * | 2015-12-24 | 2016-03-16 | 山东博科生物产业有限公司 | Glucose detection kit |
| CN106093426A (en) * | 2016-05-31 | 2016-11-09 | 安徽伊普诺康生物技术股份有限公司 | A kind of test kit measuring bladder chalone C and preparation method thereof |
-
2017
- 2017-09-30 CN CN201710917819.8A patent/CN107741491A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101324581A (en) * | 2007-06-13 | 2008-12-17 | 苏州艾杰生物科技有限公司 | Method for determining galactose concentration and galactose diagnosis/determination reagent kit |
| CN105400862A (en) * | 2015-12-24 | 2016-03-16 | 山东博科生物产业有限公司 | Glucose detection kit |
| CN106093426A (en) * | 2016-05-31 | 2016-11-09 | 安徽伊普诺康生物技术股份有限公司 | A kind of test kit measuring bladder chalone C and preparation method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109557088A (en) * | 2019-01-11 | 2019-04-02 | 四川沃文特生物技术有限公司 | A kind of reagent card for detecting excrement lactose content and the excrement lactose detection method based on this |
| CN109991222A (en) * | 2019-04-29 | 2019-07-09 | 徐詹程 | A kind of infant urine galactolipin instrument and its detection method |
| CN118243646A (en) * | 2024-04-24 | 2024-06-25 | 武汉简为医疗科技有限公司 | Lactose intolerance detection kit and application method thereof |
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Application publication date: 20180227 |
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