CN108982885A - The kit of alpha-amylase in a kind of measurement serum - Google Patents
The kit of alpha-amylase in a kind of measurement serum Download PDFInfo
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- CN108982885A CN108982885A CN201710412228.5A CN201710412228A CN108982885A CN 108982885 A CN108982885 A CN 108982885A CN 201710412228 A CN201710412228 A CN 201710412228A CN 108982885 A CN108982885 A CN 108982885A
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- G—PHYSICS
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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
- G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
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Abstract
The present invention provides a kind of assay kits of alpha-amylase in measurement serum, this kit is liquid double reagent.It is made of reagent 1 (R1) and reagent 2 (R2), R1 is mainly made of hydroxyethyl piperazine second thiosulfonic acid (HEPES), alpha-glucosidase, sodium chloride, calcium chloride, cerous nitrate；R2 is mainly made of 4,6- ethylidene -4- nitrophenols-α-heptose (E-PNP-G7), cerous nitrate.Cerous nitrate is added in the assay kit of alpha-amylase in serum provided by the invention, the performance for further improving kit, improves the reactivity of alpha-amylase, kit of the present invention and gold-labeled kit testing result correlation are good, this stabilization of kit is good, is able to satisfy the requirement of clinical examination.
Description
Technical field
The invention belongs to medicine vitro diagnostic techniques field, it is specifically related to a kind of measurement reagent of alpha-amylase in serum
Box.
Background technique
Pancreas is included there are many a large amount of digestive ferment, and in clinical position, most important have 3 kinds of pancreatin, i.e., trypsase,
Lipase and amylase.When pancreas function obstacle, one side Secretion obstacle, the amount for causing enzyme to drain to duodenum is reduced,
And the enzyme amount on the other hand escaped into blood increases, and increases the enzyme amount in blood and urine.It is different according to amylorrhexis product
The difference of structure type can be divided into alpha-amylase and beta amylase, and alpha-amylase is present in animal and microorganism, beta amylase master
It inquires in higher plant, but is also reported in bacterium, cow's milk, exists in mould.
In human body alpha-amylase not only pancreas illness, non-pancreas illness, salivary gland illness diagnosis in it is valuable, and
And be abdomen illness, macroamylasemia etc. diagnosis on important indicator.More commonly used in clinical examination is serum alphalise starch
Enzymatic determination.
So far, the method for alpha-amylase is measured up to 200 kinds or more, is mainly had:
1. using starch as total alpha-amylase activity measuring method of substrate
It is divided into viscosimetry, turbidimetry and iodine-starch in color method, wherein viscosimetry is due to technical tired
Difficulty, accuracy and precision are poor, deactivate already.There are substrate unstable the shortcomings that being difficult to marking for turbidimetry.Iodine-starch is in
Color method system semi-quantitative method cannot accurately reflect the height of enzymatic activity, and sensibility is low, easily leads to clinic and fails to pinpoint a disease in diagnosis.The method is by me
The Ministry of Public Health of state explicit order is eliminated.
2. alpha-amylase dry chemical measuring method
The method due to have the characteristics that by fast bed measurement it is easy it is accurate, accurate, practical be suitable for clinic, but due to
The degree of awareness is low, the factors such as instrument price and reagent price height, it is difficult to universal.
3. using Fructus Hordei Germinatus oligose as the alpha-amylase activity measuring method of substrate
Including direct measuring method and using Fructus Hordei Germinatus oligose as the enzyme coupled assay of substrate.Wherein direct measuring method has pollution
The disadvantages of environment.Using Fructus Hordei Germinatus oligose as the enzyme coupled assay of substrate include malt pentose for substrate, maltotetraose be substrate with
And seven sugar of malt is the enzyme coupled assay of substrate, wherein malt pentose is that substrate method application tool enzyme is more, and haemolysis is to survey
Determine result to be affected；Maltotetraose is that the intermediate products such as substrate method endogenous maltose and glucose have interference to reaction；Wheat
The sugar of bud seven is substrate method, its advantage is that it increases percent hydrolysis and closest to the affinity and starch of alpha-amylase isodynamic enzyme, this
There is the reactive problems that may change starch enzyme-to-substrate because of chemical modification for class method, cannot comply fully with IFCC formulation
With reference to method standard, but it is compared with other methods compared with, the method closest to standard.
Therefore, it is substrate method enzyme coupled assay that it is sugared, which to develop a kind of improvement malt seven, by the present invention, solves amylase and bottom
The reactive problem of object makes it meet the standard of the reference method of IFCC formulation as far as possible, is that can solve current clinical application hardly possible
Topic.
Summary of the invention
In order to overcome the drawbacks of the prior art, although patent 201410401270.3 joined dodecyl in kit
Benzene sulfonic acid sodium salt improves kit performance, but the present inventor is it is found through experiment that cerous nitrate can preferably improve amylase
The performance of detection kit improves detection sensitivity, further reduction minimum detection limit concentration, therefore is added in the present invention
Cerous nitrate further improves the performance of alpha-amylase detection kit.
Specific embodiment
Specifically, the present invention relates to following aspect, more particularly it relates to alpha-amylase in a kind of serum
Assay kit, the kit are liquid double reagent, are made of reagent 1 (R1) and reagent 2 (R2), the R1 includes with the following group
Point:
The R2 includes following components:
4,6- ethylidene -4- nitrophenols-α-heptose (E-PNP-G7) 0.1~10mmol/L
0.1~1mmol/L of cerous nitrate
Preferably, the R1 includes following components:
The R2 includes following components:
E-PNP-G7 5mmol/L
Cerous nitrate 0.5mmol/L
Specific embodiment
Below by specific embodiment, the present invention is described in detail, but the purposes of these exemplary embodiments and
Purpose is only used to enumerate the present invention, not constitutes any type of any restriction to real protection scope of the invention, more non-to incite somebody to action
Protection scope of the present invention is confined to this.
Embodiment 1
The assay kit of alpha-amylase in serum, which is liquid double reagent, by reagent 1 (R1) and reagent 2
(R2) it forms, R1 component are as follows:
R2 component are as follows:
E-PNP-G7 5mmol/L
Cerous nitrate 0.5mmol/L
Embodiment 2
The assay kit of alpha-amylase in serum, which is liquid double reagent, by reagent 1 (R1) and reagent 2
(R2) it forms, R1 component are as follows:
R2 component are as follows:
E-PNP-G7 10mmol/L
Cerous nitrate 1.0mmol/L
Kit described in embodiment 1-2, suitable for have respective wavelength 405nm, 37 DEG C of thermostats semi-automatic or
Automatic clinical chemistry analyzer.
Embodiment 3
One, the reactive verification test of starch enzyme-to-substrate can be improved by improving formula
Minimum detection limit: minimum detection limit determines use with lot number reagent to zero-dose calibration object (or sample diluting liquid)
It carries out at least 20 times repetitions to detect, the SD that average value subtracts 2 times adds the lowest detection of the i.e. reagent of 2 times of SD or more in zero point average value
Limit.
1, grinding certainly for formula described in embodiment 1 is matched described in alpha-amylase assay kit and patent 201410401270.3
The alpha-amylase assay kit of side carries out minimum detection limit verification test, compares the reaction of amylase in kit in two inventions
Property.As a result as follows:
2, grinding certainly for formula described in embodiment 2 is matched described in alpha-amylase assay kit and patent 201410401270.3
The alpha-amylase assay kit of side carries out minimum detection limit verification test, compares the reaction of amylase in kit in two inventions
Property.As a result as follows:
It in summary it can be seen, autogamy embodiment 1-2 detection limit of the present invention is respectively 5.061U/L and 5.105U/L, and patent
The 201410401270.3 alpha-amylase assay kit detection limit 11.026U/L for improving formula, the present invention is compared with patent
The 201410401270.3 improvement formulas have more preferably improvement to amylase reactivity, can further improve detection
The sensitivity of kit, therefore the present invention improves the determination of amylase kit of formula, the enhancing of amylase reactivity effect is sensitive
Degree is further enhanced.So that the assay kit detection method of alpha-amylase more accords in serum of the invention
The standard for closing the reference method that IFCC is formulated, solves current clinical application problem, it will has more actual clinical value.
Embodiment 4
Two, measurement Accuracy Verification test
1, using the alpha-amylase of 1 kit of the embodiment of the present invention and Shanghai Kehua Bio-engineering Co., Ltd's production
Kit is tested using the relevance verification that Beckman DxC600 automatic clinical chemistry analyzer carries out the two, takes 50 patient's blood
It is tested clearly, the correlation of both comparisons testing result.
The results show that grind certainly the alpha-amylase assay kit and Shanghai China of section bioengineering share of autogamy embodiment 1 have
The gold-labeled kit consistency of performance of limit company production is good.
2, using the alpha-amylase of 2 kit of the embodiment of the present invention and Shanghai Kehua Bio-engineering Co., Ltd's production
Kit is tested using the relevance verification that Beckman DxC600 automatic clinical chemistry analyzer carries out the two, takes 50 patient's blood
It is tested clearly, the correlation of both comparisons testing result.
The results show that grind certainly the alpha-amylase assay kit and Shanghai China of section bioengineering share of autogamy embodiment 2 have
The gold-labeled kit consistency of performance of limit company production is good.
Embodiment 5
Three, the stability verification test of this kit
1, the stability of the kit of example 1 group point is investigated
The alpha-amylase assay kit of formula described in embodiment 1 is carried out to stability to examine under 2~8 DEG C of conditions of storage
Core, the situation of change of detection assay value, every three months detection is primary, until 13 months.
2, the stability of the kit of 2 component of embodiment is investigated
The alpha-amylase assay kit of formula described in embodiment 2 is carried out to stability to examine under 2~8 DEG C of conditions of storage
Core, the situation of change of detection assay value, every three months detection is primary, until 13 months.
The above result shows that 2 DEG C~8 DEG C are stored 13 months, measured value (measured value and target within the tolerance of target value
The deviation of value should be ± 10%), alpha-amylase assay kit of the present invention has good stability.
In summary information, of the invention alpha-amylase assay kit of grinding in serum certainly on the basis of improving formula,
It is able to maintain good with gold-labeled kit testing result consistency, testing result is accurate and reliable, and kit stores under condition of storage
13 months, performance was stablized, and has more preferably improvement to amylase reactivity, can further improve detection kit
Sensitivity, therefore the determination of amylase kit of improvement formula of the invention, the enhancing of amylase reactivity effect, sensitivity into
One step is improved.So that the assay kit detection method of alpha-amylase more meets in serum of the invention
The standard for the reference method that IFCC is formulated, solves current clinical application problem.Actual clinical value will be had more.
It should be appreciated that the purposes of these embodiments is merely to illustrate the present invention and is not intended to limit protection model of the invention
It encloses.In addition, it should also be understood that, after reading the technical contents of the present invention, those skilled in the art can make the present invention each
Kind change, modification and/or variation, all these equivalent forms equally fall within guarantor defined by the application the appended claims
Within the scope of shield.
Claims (2)
1. the assay kit of alpha-amylase in a kind of serum, which is characterized in that the kit is liquid double reagent, by reagent 1
(R1) it is formed with reagent 2 (R2), the R1 includes following components:
The R2 includes following components:
4,6- ethylidene -4- nitrophenols-α-heptose (E-PNP-G7) 0.1~10mmol/L
0.1~1mmol/L of cerous nitrate.
2. the assay kit of alpha-amylase in serum as described in claim 1, which is characterized in that the kit is that liquid is double
Reagent is made of reagent 1 (R1) and reagent 2 (R2), and the R1 includes following components:
The R2 includes following components:
E-PNP-G7 5mmol/L
Cerous nitrate 0.5mmol/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710412228.5A CN108982885B (en) | 2017-06-05 | 2017-06-05 | Kit for measuring alpha-amylase in serum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710412228.5A CN108982885B (en) | 2017-06-05 | 2017-06-05 | Kit for measuring alpha-amylase in serum |
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| Publication Number | Publication Date |
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| CN108982885A true CN108982885A (en) | 2018-12-11 |
| CN108982885B CN108982885B (en) | 2021-11-16 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1912580A (en) * | 2005-08-08 | 2007-02-14 | 苏作军 | Serum starch enzyme reagent kit and preparation method thereof |
| EP1342790B1 (en) * | 2002-03-07 | 2009-06-24 | Toyama University | Method for measuring amylase activity |
| CN104198691A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Stable alpha-amylase detection kit |
| CN105021543A (en) * | 2015-06-30 | 2015-11-04 | 广州金域医学检验中心有限公司 | Alpha-amylase detection reagent and application thereof |
-
2017
- 2017-06-05 CN CN201710412228.5A patent/CN108982885B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1342790B1 (en) * | 2002-03-07 | 2009-06-24 | Toyama University | Method for measuring amylase activity |
| CN1912580A (en) * | 2005-08-08 | 2007-02-14 | 苏作军 | Serum starch enzyme reagent kit and preparation method thereof |
| CN104198691A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Stable alpha-amylase detection kit |
| CN105021543A (en) * | 2015-06-30 | 2015-11-04 | 广州金域医学检验中心有限公司 | Alpha-amylase detection reagent and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 苏国均等: "Ce3+对大鼠体内淀粉酶同工酶的影响", 《中山大学学报（自然科学版）》 * |
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| CN108982885B (en) | 2021-11-16 |
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Inventor after: Zhang Chuanchun Inventor after: Qu Xingcui Inventor after: Bai Lijuan Inventor after: Tan Zaili Inventor after: Luo Yuxiang Inventor before: Zhang Chuanchun Inventor before: Qu Xingcui |
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