CN109724933A - A kind of reproducible aspartate amino transferase detection kit - Google Patents
A kind of reproducible aspartate amino transferase detection kit Download PDFInfo
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- CN109724933A CN109724933A CN201811653698.1A CN201811653698A CN109724933A CN 109724933 A CN109724933 A CN 109724933A CN 201811653698 A CN201811653698 A CN 201811653698A CN 109724933 A CN109724933 A CN 109724933A
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Abstract
The present invention relates to biochemistry detection technical field, in particular to a kind of reproducible aspartate amino transferase detection kit and its preparation, application method.It includes reagent R1 and reagent R2, and wherein reagent R1 includes buffer, pH value adjustment agent, ASPARTIC ACID, potassium ferrocyanide, AP5A, EGTA, lithium chloride, surfactant, left-handed glucosides, lactic dehydrogenase, malic dehydrogenase, preservative component, reagent R2 include buffer, pH value adjustment agent, BSA, preservative, a-ketoglutaric acid, NADH component.The present invention is optimized from removal reflection interference, the activity for improving the enzyme reacted, the homogeneity for improving reaction system and preferably excellent commplementary wave length etc., the present invention significantly improves the repeatability of reagent, the especially repeatability of low value sample, to be conducive to improve the accuracy of clinical diagnosis result, there is very big value for clinical application.
Description
Technical field
The invention belongs to biochemistry detection technical fields, and in particular to a kind of reproducible aspartate amino transferase
Detection kit and its preparation, application method.
Background technique
Aspartate amino transferase (aspartate aminotransferase, AST), is present in various groups of human body
It is most abundant with levels and myocardial contents in knitting, followed by liver.Under normal circumstances, content of the AST in serum is very low, works as linked groups
When organ damages, concentration of the AST in serum can occur to change accordingly.
Liver blood detection is the most common blood testing, can be used for assessing liver function or hepatic injury.Chronic hepatitis is suffered from
The most common laboratory of person checks that abnormal is that serum transaminase increases, common serum aspartate transaminase (AST).AST master
It is distributed in liver cell, sub-fraction is present in muscle cell.Transaminase if liver damage or damage, in liver cell
Just enter blood, AST level increases in blood, prompts liver diseases signal.Wherein AST is most sensitive, it is in hepatic tissue
Activity is 100 times in serum, as long as there is 1% necrosis of liver cells, the AST in serum can be made to increase by 1 times.Therefore, most of
In the case of AST elevated-levels it is consistent with the liver cell extent of damage, be most common Liver function grade index.
The measurement of serum aspartate amino transferase facilitates the judgement heart and liver cell, and whether there is or not necrosis and degree of injury.
AST increases: being common in acute and chronic serious hepatitis, cirrhosis, myocarditis, myocardial infarction, ephritis, cholangitis, dermatomyositis, pancreas
The illnesss such as inflammation.AST is reduced: over fatigue, the diseases such as malnutritive and serious later period of hepatocarcinoma.
The detection method of AST uses ultraviolet spectrophotometry mainly in combination with automatic clinical chemistry analyzer device in clinical application
To the content of the AST in human serum or blood plasma by way of continuously detecting, testing principle is main are as follows:
ASPARTIC ACID and α-ketoglutaric acid generate oxaloacetic acid and Pidolidone, oxalyl second under AST enzymatic reaction
Acid and reduced coenzyme Ⅰ (NADH) generate L MALIC ACID and cozymase, reduction under the catalytic action of malic dehydrogenase (MDH)
The rate of type cozymase (NADH) consumption and the activity of sample Mid-Heaven Gate aspartic acid aminopherase (AST) are directly proportional, pass through measurement
The activity of sample Mid-Heaven Gate aspartic acid aminopherase (AST) can be obtained in the change rate of absorbance at 340nm.
The detection method has at low cost, the advantages such as mass detection speed is fast, accuracy is good, at present in clinic
It is widely applied, but this method is often repeated very undesirable for the low value sample occurred in clinic, in some precision
In slightly worse instrument and equipment, the problem of poor repeatability of performance, can more be protruded, and the sample of normal person is generally concentrated at 12-
In the range of 40U/L, and tool is diagnosed lower than patient of the patient's sample of normal reference range for liver tumour advanced stage in clinic
There is important reference significance, and the sample in normal reference range is relatively low, therefore improves this method product
Repeatability also has very important practical value.
For this problem, the reason that examen causes repeatability undesirable is invented a kind of with good reproducibility
Aspartate amino transferase (AST) detection kit and application method, utilize clinical popularization and use.
Summary of the invention
To solve the above problems, the purpose of the present invention is to provide a kind of reproducible aspartate amino transferases
Detection kit, using the method continuously detected based on uv-spectrophotometric, by the interfering substance of removal interference repeatability,
It improves the homogeneity of reaction system and improves the methods of the sensitivity of enzyme reaction, invent a kind of reproducible asparatate
Transamination enzyme detection kit and its preparation, application method.
To achieve the above object, the present invention is achieved by the following technical solutions: a kind of reproducible asparagine
Sour transamination enzyme detection kit comprising reagent R1 and reagent R2,
The reagent R1 includes the component of following content:
The reagent R2 includes the component of following content:
A kind of reproducible aspartate amino transferase detection kit, it is characterised in that: buffered in reagent R1
Liquid is 25 DEG C, pH are as follows: 7.4 Tris buffer；Buffer is 25 DEG C in reagent R2, the CAPS buffer that pH is 9.8.
A kind of reproducible aspartate amino transferase detection kit, it is characterised in that: the pH value adjustment
Agent is lithium hydroxide or tartaric acid.
A kind of reproducible aspartate amino transferase detection kit, it is characterised in that: the preservative is
Polylysine.
A kind of reproducible aspartate amino transferase detection kit, it is characterised in that: the surface-active
Agent 1 is that Qula leads to -108 (SIGMA)), surfactant 2 is oleamide propyl betaine (Japanese Kao).
A kind of application method of reproducible aspartate amino transferase detection kit, it is characterised in that: make
Be measured with automatic clinical chemistry analyzer using end-point method, detection dominant wavelength be 340nm, a length of 546nm of complementary wave, R1 reagent and
The ratio of R2 reagent is 4:1.
The beneficial effects of the present invention are:
1) this law is bright according to preferred Tris buffer in reagent R1 the characteristics of enzyme in reagent and substrate, in reagent R2 preferably
CAPS buffer, and the preferred lithium hydroxide of purpose PH required for adjusting and tartaric acid are as regulator, buffer and PH
The activity of value regulator preferably facilitated etc. than improving catalysis reaction, utilizes the raising of repeatability；
2) present invention increase AP5A assists in removing interference of the kinases to reaction in dry blood, and potassium ferrocyanide is added with
Help remove interference of the fluctuation of bilirubin to continuous detection method, the repeatability of detection reagent is improved in terms of removing chaff interferent；
3) two kinds of surfactant Qulas logical -108 and oleamide propyl betaine be advantageously selected for reaction solution is fast
The homogenization of speed, improves the uniformity of detection reaction every time, and reaction solution is promoted more to clarify, and increases the permeability of optical path, from
And improve the repeatability of detection detected every time；
4) the preferred EGTA metal-chelator of property in the present invention also according to key tool enzyme in reagent, reduces heavy metal
The inhibiting effect of ions enzyme, preferably polylysine effectively inhibit as preservative the growth of bacterial micro-organism in reagent,
Increase the protective agents such as left-handed glucosides, bovine serum albumin(BSA) in reagent come property the problem of improving enzyme, guarantees that enough enzymatic activitys participate in
Reaction.
5) present invention preferably 546nm can be interfered as commplementary wave length caused by effective compensation apparatus fluctuation, and to detection
As a result it will not have an impact.
Specific embodiment
In order to be easy to understand the technical means, the creative features, the aims and the efficiencies achieved by the present invention, tie below
Closing specific embodiment, invention is further explained:
Embodiment 1
1) detection reagent of the common aspartate amino transferase of one kind provided by the present embodiment (AST), including
Reagent R1 and reagent R2, concrete composition ingredient:
Reagent R1:
Reagent R2:
2) application method of the present embodiment reagent:
Common aspartate amino transferase (AST) detection kit of one kind of the present embodiment description, when in use
Using the automatic clinical chemistry analyzer with double reagent function, such as 7180 fully-automatic analyzer of Hitachi, Rate A method is utilized
It is measured.R1 and R2 is placed on corresponding reagent position according to the ratio of 4:1, places steaming in the corresponding position of sample disc
Distilled water, standard items and sample, operation such as table 1.
1 reagent test method table of table
It calculates: aspartate amino transferase (AST) content (U/L) with good reproducibility=(Δ A measures ÷ Δ
A standard) × C standard
Embodiment 2
The reproducible aspartate amino transferase detection kit of one kind described in the present embodiment, packet reagent R1
With reagent R2:
1) component of reagent R1 includes:
The reagent R2 includes the component of following content:
2) application method of the present embodiment reagent:
The present embodiment reagent is when in use using the automatic clinical chemistry analyzer with double reagent function, as Hitachi 7180 is complete
Automatic analyzer etc. is measured using Rate A method.R1 and R2 is placed into corresponding reagent position according to the ratio of 4:1
On, distilled water, standard items and sample, operation such as table 2 are placed in the corresponding position of sample disc.
2 reagent test method table of table
It calculates: aspartate amino transferase (AST) content (U/L) with good reproducibility=(Δ A measures ÷ Δ
A standard) × C standard
Embodiment 3
The reproducible aspartate amino transferase detection kit of one kind described in the present embodiment, packet reagent R1
With reagent R2:
1) component of reagent R1 includes:
The reagent R2 includes the component of following content:
2) application method of the present embodiment is the same as embodiment 2.
Repeated performance verification:
Repetitive test: choosing high level, intermediate value, low value, each two groups of clinical sample of four different stages of extremely low value respectively,
Control test is carried out with embodiment 1-3 respectively, 20 detections are carried out to every part of sample, totally 20 testing results will be calculated averagely
Value, standard deviation and the coefficient of variation.It the results are shown in Table 3.
Table 3-1 high level sample repeatability testing result
Table 3-2 intermediate value sample repeatability testing result
Table 3-3 low value sample repeatability testing result
The extremely low value sample repeatability testing result of table 3-4
By the CV% coefficient of variation in table 3 it is found that compared with comparative example 1-3, embodiment 2 and the coefficient of variation implemented in 3 are equal
Smaller, especially in terms of low value sample and extremely low value sample, testing result has preferably repeatability, this illustrates the present invention
By adjusting reaction system, increases the methods of anti-interference agent, optimizes reaction process, greatly improve the repeatability of reagent,
The especially repeatability of low value sample, effect is clearly.
Claims (6)
1. a kind of reproducible aspartate amino transferase detection kit, it is characterised in that: it include reagent R1 and
Reagent R2,
The reagent R1 includes the component of following content:
The reagent R2 includes the component of following content:
2. the reproducible aspartate amino transferase detection kit of one kind according to claim 1, feature
It is: trishydroxymethylaminomethane (Tris) buffer that buffer is 25 DEG C in reagent R1, pH is 7.4；It is buffered in reagent R2
Liquid is 25 DEG C, 3- (cyclohexylamine) -1- propane sulfonic acid (CAPS) buffer that pH is 9.8.
3. the reproducible aspartate amino transferase detection kit of one kind according to claim 1, feature
Be: the pH value adjustment agent is lithium hydroxide or tartaric acid.
4. the reproducible aspartate amino transferase detection kit of one kind according to claim 1, feature
Be: the preservative is polylysine.
5. the reproducible aspartate amino transferase detection kit of one kind according to claim 1, feature
Be: the surfactant 1 is that Qula leads to -108 (SIGMA)), surfactant 2 is oleamide propyl betaine (Japan
Flower king).
6. the reproducible aspartate amino transferase detection reagent of one kind described in -5 any one according to claim 1
The application method of box, it is characterised in that: be measured using automatic clinical chemistry analyzer using end-point method, detection dominant wavelength is
The ratio of 340nm, a length of 546nm of complementary wave, R1 reagent and R2 reagent is 4:1.
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| CN201811653698.1A CN109724933A (en) | 2018-12-30 | 2018-12-30 | A kind of reproducible aspartate amino transferase detection kit |
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| CN201811653698.1A CN109724933A (en) | 2018-12-30 | 2018-12-30 | A kind of reproducible aspartate amino transferase detection kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115655848A (en) * | 2022-12-26 | 2023-01-31 | 河北盛华尔生物医疗科技有限公司 | Stable glutamic-oxaloacetic transaminase determination kit |
| WO2023060818A1 (en) * | 2021-10-12 | 2023-04-20 | 广州达安基因股份有限公司 | In-vitro diagnostic reagent preservative and use thereof |
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