CN106282312A - A kind of dry chemistry bigeminy reagent strip for detecting glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase simultaneously - Google Patents
A kind of dry chemistry bigeminy reagent strip for detecting glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase simultaneously Download PDFInfo
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- CN106282312A CN106282312A CN201610692719.5A CN201610692719A CN106282312A CN 106282312 A CN106282312 A CN 106282312A CN 201610692719 A CN201610692719 A CN 201610692719A CN 106282312 A CN106282312 A CN 106282312A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
The invention discloses a kind of dry chemistry bigeminy reagent strip for detecting glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase simultaneously.It is characterized in that this bigeminy reagent strip comprises two sections of test paper pads, each test paper pad is formed by four tunics, is top layer diffusion barrier, second layer rapid glass fibrous filter membrane, third layer composite fibre film and bottom reaction film respectively.It is adsorbed with corresponding reactant on reaction film, after adding detected material in sample holes, by diffusion into the surface film so that detected material evenly spreads on two panels test paper pad, eventually arrives at bottom reaction film and react.Meanwhile, reagent strip lower supporting layer has two instrument connections, is connected with two reaction films of bottom respectively, in inverse process, absorption dihydrocoenzyme I (NADH) on reaction film is oxidized to NAD+, the fall of the absorptance at test 340nm, thus obtains the content of corresponding detected material.This reagent strip can be used for quickly measuring the activity of glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase in human whole blood, serum or blood plasma, thus the beneficially clinical monitoring of hepatic disease and auxiliary treatment.
Description
Technical field
The present invention relates to clinical vitro detection technical field, be specifically related to detect glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase simultaneously
Method.
Background technology
Glutamic oxaloacetic transaminase, GOT (AST) and glutamate pyruvate transaminase (ALT), be primarily present in various cell, especially with hepatocyte for,
Time normal, it is present in the most on a small quantity in blood.In the acute stage of various viral hepatitis, during drug intoxication hepatic necrosis, AST
Being released in a large number in blood with ALT, therefore both transaminases are the important indicators of Diagnosis Virus Hepatitis, toxic hepatitis.With
Time be also the highstrung index of hepatitis patient, transaminase is the highest, and the general reflection impaired degree of hepatocyte is the most serious, and from certain
Point out in kind of degree the state of an illness weight and more after.AST/ALT ratio is determined with certain clinic to hepatic fibrosis degree and refers to
Lead meaning.Medical findings shows, AST/ALT ratio is gradually increasing along with increasing the weight of of degree of hepatic fibrosis, works as AST/ALT
During ratio >=1, diagnosis early stage liver cirrhosis had stronger directive significance.
At present, the method for external test AST and ALT mainly uses continuous monitoring method, the method be international clinical chemistry and
The recommendation method of laboratory medicine alliance (IFCC).Concrete mensuration process is as follows:
1. continuous monitoring method detection glutamic oxaloacetic transaminase, GOT (AST)
2. continuous monitoring method detection glutamate pyruvate transaminase (ALT)
Above-mentioned two groups of reactions all cause the minimizing of NADH, cause absorbance to decline at wavelength 340nm, fall off rate and two
Plant transferring enzyme (AST and ALT) vigor to be directly proportional.
The reagent being used for detecting AST and ALT clinically can be divided into liquid reagent and drying chemical reagent paper two class.Same liquid reagent
Compare, chemical reagent bar has conveniently, flexibly, pollute the advantages such as little, easy and simple to handle.
Now mostly armarium is to separate two kinds of indexs of AST and ALT to detect, the most time-consuming but also waste resource,
It is desirable to provide one detects the dry chemistry bigeminy reagent strip of glutamic oxaloacetic transaminase, GOT (AST) and glutamate pyruvate transaminase (ALT) simultaneously.
Summary of the invention
Present disclosure be to provide a kind of easy to use, highly sensitive can Simultaneous Determination human whole blood, blood
Glutamic oxaloacetic transaminase, GOT (AST) clearly or in blood plasma and the dry chemistry bigeminy reagent strip of glutamate pyruvate transaminase (ALT).
The dry chemistry bigeminy reagent strip of quantitative determination AST and ALT that the present invention provides, for strip, one end is holding area,
The other end is test section, its structure such as Fig. 1 and 2, and it is by upper supporting layer 1, lower supporting layer 2, diffusion barrier 3, hemofiltration film 4,5 and reaction
Film 6 forms.Have well 7 in the test section of upper supporting layer 1, the correspondence position of lower supporting layer 2 has instrument connection 8 and 9.
In the present invention, reagent strip shell upper supporting layer 1 and lower supporting layer 2 use macromolecule polymeric material to make, and thickness is
0.5mm, upper strata size is 22 × 10mm, and lower floor's size is 45.5 × 10mm.Well and instrument connection on upper lower supporting layer are
Circle, internal diameter is identical, for 4mm.Well 7 is used for dripping sample, and instrument connection 8 and 9 is for measuring the change of reaction zone absorbance.
In the present invention, diffusion barrier 3 is used for accepting sample and spreading downwards, and preferably reaction zone area can accommodate about 10-30 μ
L blood, by two layers of blood separating films, and can be transported to bottom reaction film by test sample product by about 6-20 μ L.
In the present invention, the reaction scheme of employing is enzyme coupling continuous monitoring method, in this coupling reaction, and the oxidation speed of NADH
Rate with specimen enzymatic activity to be measured proportional, at 340nm, monitor the rate of descent of absorbance, thus draw the activity of AST and ALT
Unit.
In the present invention, comprise two groups of reaction systems, be respectively used to detect AST and ALT.For detecting the reaction system of AST
Including: TRIS buffer, concentration 280mM, pH is 7.8；Substrate is L-Aspartic acid salt and sodium alpha-ketoglutarate, wherein L-Radix Asparagi
The concentration of propylhomoserin salt is 0.8M-1.5M；The concentration of sodium alpha-ketoglutarate is 90mM-120mM；The concentration of lactic acid dehydrogenase (LDH)
For > 3.0KU/L；The concentration of malic dehydrogenase (MDH) is > 2.5KU/L；The concentration of NADH > 2mM；The stabilizer of enzyme is Sanguis Bovis seu Bubali
Pure albumen weight concentration in system is 0.1%-0.25%；Triton X-100 and Hydrazoic acid,sodium salt weight concentration are
0.01%.Including for detecting the reaction system of ALT: TRIS buffer, concentration 250mM, pH is 7.5；Substrate is ALANINE
And sodium alpha-ketoglutarate, wherein the concentration of ALANINE is 1M-1.5M；The concentration of sodium alpha-ketoglutarate is 50mM-150mM；Breast
The concentration of acidohydrogenase (LDH) is > 2KU/L；The concentration of NADH > 2mM；The stabilizer of enzyme is that bovine serum albumin is in system
Weight concentration is 0.1%-0.25%；Triton X-100 and Hydrazoic acid,sodium salt weight concentration are 0.01%.
For quantitative determination AST and ALT activity, inventor's unit still further developed and the two matching used reflective light splitting of bracing
Photometer (separate case is applied for a patent).30-60 μ L sample of blood drips in reagent strip from well, and reaction temperature controls in room temperature, surveys
Examination completed in 3-5 minute, instrument automatically report and store experimental result.
The invention has the beneficial effects as follows: the present invention uses the technology of dry type reagent strip to achieve simultaneously and detects glutamic oxaloacetic transaminase, GOT
With the sensitive indicator of glutamate pyruvate transaminase the two liver, by detection that the index with clinical meaning is combined, greatly
Simplify the formality of seeking medical advice of patient, shorten the time that doctor makes a definite diagnosis, significantly improve diagnosis efficiency；The bigeminy of present invention design
The enzyme coupling continuous detecting method that the universal degree of recognition of strip adoption is higher, reacts rapid sensitive, and diagnostic result is the most relatively reliable；The present invention
With the addition of bovine serum albumin in detectable, the active group of protection LDH is not destroyed, it is possible to be effectively improved LDH raw
Change the stability of reagent, thus ensure that the stability of two bracings.
Accompanying drawing explanation
Fig. 1 is that the dry chemistry reagent bar overall structure of the present invention resolves diagram.
Fig. 2 is that the dry chemistry reagent bar shell of the present invention launches and folds diagram.
Label in figure: 1 is upper supporting layer, 2 is lower supporting layer, and 3 is diffusion barrier, and 4 is rapid glass fibrous filter membrane, and 5 is mixed
Condensating fiber film, 6 is LP reaction film, and 7 is well, and 8 is glutamic oxaloacetic transaminase, GOT instrument connection, and 9 is glutamate pyruvate transaminase instrument connection.
Detailed description of the invention
On the lower supporting layer 2 of Fig. 1, overlapping diffusion barrier 3, rapid glass fibrous filter membrane 4, composite fibre film 5, LP in order
Reaction film 6, then upper supporting layer 1 is pressed on lower supporting layer 2 fixing.
The size of every tunic controls at about 6 × 6mm, after pressing is fixing, it is ensured that between two sections of each tunics of test strips especially
Being not have adhesive band between bottom reaction film, instrument connection corresponds to reaction film center, must not have deviation, otherwise by impact detection knot
Really.
It is as follows that the present invention carries out the detection method of AST and ALT in blood sample:
1. add sample: inserted by reagent strip in matching used instrument, then by fluid sample or treated
Fluid sample drops in reagent strip well of the present invention；
2. chromatography permeable reactive: stand several minutes and wait that sample chromatography process of osmosis completes and contacts with bottom reaction film,
Carry out enzyme reaction；
3. result interpretation: due to the fact that and fixed by reaction film, thereby through instrument to optics signal change on film
Measure, interpretation can be realized, thus realize the qualitative and quantitative detection to detected material AST and ALT and analysis.
Claims (5)
1. for detection glutamic oxaloacetic transaminase, GOT and the dry chemistry bigeminy reagent strip of glutamate pyruvate transaminase simultaneously, including reagent strip shell and interior
Layer test strips, it is characterised in that reagent strip shell upper supporting layer offers well, and lower supporting layer offers two instrument connections, point
Not corresponding two test events.
Dry chemistry bigeminy reagent for detecting glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase simultaneously the most according to claim 1
Bar, it is characterised in that internal layer test strips is made up of four tunics, respectively is diffusion barrier, hemofiltration film (two-layer) and reaction film, no
With the corresponding different reaction film of detectable substance, on it, the reacting substance of absorption is different.
Internal layer test strips the most according to claim 2, it is characterised in that top layer diffusion layer be one layer uniform each to equal property
Porous diffusion film, wet-film thickness is 100-300 μm；Two layers of blood separating films are rapid glass fibrous filter membrane the most respectively and mix
Condensating fiber film；Bottom reaction film is one layer of LP film, and it is adsorbed with corresponding reaction reagent.
4. according to the bottom reaction film described in claim 2 and 3, it is characterised in that different detection projects is adsorbed with different anti-
Answer reagent.Including buffer system, TRIS buffer, concentration 280mM for detecting the reaction reagent of glutamic oxaloacetic transaminase, GOT, pH is 7.8；
Substrate is L-Aspartic acid salt and sodium alpha-ketoglutarate, and wherein the concentration of L-Aspartic acid salt is 0.8M-1.5M；α-one penta
The concentration of diacid sodium is 90mM-120mM；
The concentration of lactic acid dehydrogenase (LDH) is > 3.0KU/L；
The concentration of malic dehydrogenase (MDH) is > 2.5KU/L；
The concentration of NADH > 2mM；
Bovine serum albumin concentration in system is 0.1%-0.25%；
Triton X-100 and Hydrazoic acid,sodium salt concentration are 0.01%.
Include buffer system, TRIS buffer for detecting the reaction reagent of glutamate pyruvate transaminase, concentration be 250mM, pH be 7.5；
Substrate is ALANINE and sodium alpha-ketoglutarate, and wherein the concentration of ALANINE is 1.0M-1.5M；Sodium alpha-ketoglutarate
Concentration is 50mM-150mM；
The concentration of lactic acid dehydrogenase (LDH) is > 2KU/L；
The concentration of NADH > 2mM；
Bovine serum albumin concentration in system is 0.1%-0.25%；The concentration of Triton X-100 and Hydrazoic acid,sodium salt is
0.01%.
5. according to described in claim 1 or 4 for detection glutamic oxaloacetic transaminase, GOT and dry chemistry two joint-trial of glutamate pyruvate transaminase simultaneously
Agent bar, it is characterised in that bottom reaction film there will be the transformation of absorptance with detected material after contacting, can be divided by instrument connection
Correspondingly do not record, thus reach qualitative and quantitative detection glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase.
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|---|---|---|---|
| CN201610692719.5A CN106282312A (en) | 2016-08-19 | 2016-08-19 | A kind of dry chemistry bigeminy reagent strip for detecting glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase simultaneously |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610692719.5A CN106282312A (en) | 2016-08-19 | 2016-08-19 | A kind of dry chemistry bigeminy reagent strip for detecting glutamic oxaloacetic transaminase, GOT and glutamate pyruvate transaminase simultaneously |
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| CN106282312A true CN106282312A (en) | 2017-01-04 |
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Cited By (8)
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| CN109580927A (en) * | 2019-01-04 | 2019-04-05 | 杭州联晟生物科技有限公司 | A kind of test card of liver function and preparation method thereof |
| CN109900689A (en) * | 2019-03-28 | 2019-06-18 | 长沙中生众捷生物技术有限公司 | Interference rejection membrane and liver function joint test item |
| CN110412027A (en) * | 2019-08-29 | 2019-11-05 | 安徽九陆生物科技有限公司 | A kind of dry chemistry reagent item of simultaneous quantitative detection breast milk multi-parameter |
| CN112305215A (en) * | 2020-09-17 | 2021-02-02 | 上海泽恒生物科技有限公司 | Kit for co-immunoprecipitation recovery and application thereof |
| CN112672642A (en) * | 2018-09-24 | 2021-04-16 | 莱利专利股份有限公司 | Milking system with detection system |
| CN112924446A (en) * | 2021-01-29 | 2021-06-08 | 苏州国科华睿生物医学工程技术有限公司 | Test strip bearing plate, test paper box and analyzer for dry-type chemical detection |
| CN113588747A (en) * | 2021-07-12 | 2021-11-02 | 成都云芯医联科技有限公司 | Electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase |
| CN114292898A (en) * | 2021-12-02 | 2022-04-08 | 深圳市锦瑞生物科技股份有限公司 | Preparation method of glutamic-oxalacetic transaminase determination reagent and reagent ball and determination chip |
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Cited By (11)
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| CN112672642A (en) * | 2018-09-24 | 2021-04-16 | 莱利专利股份有限公司 | Milking system with detection system |
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| CN109900689A (en) * | 2019-03-28 | 2019-06-18 | 长沙中生众捷生物技术有限公司 | Interference rejection membrane and liver function joint test item |
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| CN110412027A (en) * | 2019-08-29 | 2019-11-05 | 安徽九陆生物科技有限公司 | A kind of dry chemistry reagent item of simultaneous quantitative detection breast milk multi-parameter |
| CN112305215A (en) * | 2020-09-17 | 2021-02-02 | 上海泽恒生物科技有限公司 | Kit for co-immunoprecipitation recovery and application thereof |
| CN112924446A (en) * | 2021-01-29 | 2021-06-08 | 苏州国科华睿生物医学工程技术有限公司 | Test strip bearing plate, test paper box and analyzer for dry-type chemical detection |
| CN113588747A (en) * | 2021-07-12 | 2021-11-02 | 成都云芯医联科技有限公司 | Electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase |
| CN113588747B (en) * | 2021-07-12 | 2024-01-19 | 成都云芯医联科技有限公司 | Electrochemical test card for simultaneously measuring glutamic pyruvic transaminase and glutamic oxaloacetic transaminase |
| CN114292898A (en) * | 2021-12-02 | 2022-04-08 | 深圳市锦瑞生物科技股份有限公司 | Preparation method of glutamic-oxalacetic transaminase determination reagent and reagent ball and determination chip |
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