CN113075139B - Stable double-reagent blood ammonia determination kit - Google Patents
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Abstract
The invention relates to a stable double-reagent blood ammonia determination kit, and belongs to the field of medical in-vitro diagnosis. According to the stable double-reagent blood ammonia determination kit, the L-glutamine synthetase (L-GS) and the glutaric acid disodium salt are applied to a reaction system, and the stability of the reagent storage process is ensured under the condition that the main reaction is not influenced. The stable double-reagent blood ammonia determination kit can eliminate the interference of impurity AMM, has good stability and is based on an enzyme dynamics method. The kit has the advantages of simple preparation process, high accuracy and good repeatability, and is suitable for in vitro determination of the ammonia content in serum and plasma samples.
Description
Technical Field
The invention belongs to the technical field of medical in-vitro diagnosis, and particularly relates to a stable double-reagent blood ammonia determination kit.
Background
Ammonia (Ammonia, AMM/NH) 3 ) Is an amino donor in human body and is an important metabolite. The normal value of blood ammonia is 18-72 μmol/L, which is synthesized mainly in the intestine and converted to urea and glutamine in the liver, which then passes through the kidneys to remove urea and ammonia. Therefore, the blood ammonia concentration is closely related to the functional status of three organs of intestine, liver and kidney. High concentration ammonia has strong toxicity to human central nervous system, and is a typical sign of liver diseases such as hepatitis and liver cirrhosis. In addition, the blood ammonia determination has important significance for the detection, curative effect and post-cure monitoring of hepatic encephalopathy coma.
Currently, ion exchange methods, diffusion methods, indophenol dye methods, dry chemical methods, enzymatic methods, and the like are used to test the ammonia content in blood or body fluid samples. The method has the advantages of high speed, high specificity and simple operation, and can be applied to various medical detection mechanisms in cooperation with a full-automatic biochemical analyzer. Currently, the main reactions for determining blood ammonia concentration by the enzyme kinetic method are that glutamate dehydrogenase (GLDH), alpha-ketoglutarate (alpha-KG), reduced Nicotinamide Adenine Dinucleotide (NADH) participate:
the ammonia content of the sample was calculated using the rate of change of absorbance at 340 nm.
In a few years ago, how to ensure the activity of GLDH by the system is a main technical difficulty, and the commonly used stable substances are as follows: (1) chelating agents: metal chelating agents such as disodium Ethylene Diamine Tetraacetate (EDTA), cyclohexanediamine tetraacetate (CDTA) and the like can be chelated with divalent and tetravalent cations to ensure that the enzyme is not interfered by the cations; (2) proteins: such as Bovine Serum Albumin (BSA), human Serum Albumin (HSA). The reagent with lower enzyme content can be added with protein protective agent to form higher concentration, so that the enzyme has higher anti-interference capability; (3) macromolecular species: such as trehalose, polyvinyl ether and the like, can form a protective film with protein molecules to achieve the effect of keeping enzyme activity. Fortunately, the GLDH extracted from animal tissues widely used in recent years has good stability (pH 6-9), and researchers can ensure the stability of the activity of the GLDH enzyme in the system only by simple treatment.
However, the reagent contains trace amount of AMM impurity, and is difficult to eliminate, and the reagent generates interference of blank AMM during the placing process, so that the positive error of the measured value is caused. At present, no enzyme kinetic method can effectively eliminate the positive error, so that an ammonia determination kit of an enzyme kinetic method, which can eliminate impurities and ammonia generated in the placement process and has good stability, is urgently needed.
Disclosure of Invention
The invention aims to solve the technical problems in the prior art and provides a stable double-reagent blood ammonia determination kit which can eliminate the interference of impurity AMM, has good stability and is based on an enzyme kinetics method.
In order to solve the technical problems, the technical scheme of the invention is as follows:
the invention provides a stable double-reagent blood ammonia determination kit, which comprises a reagent R1 and a reagent R2;
the reagent R1 consists of:
buffer solution with pH of 1, 8.0-9.0 and concentration of 50-200mmol/L;
alpha-ketoglutaric acid, the concentration is 1-10mmol/L;
glutaric acid disodium salt, the concentration is 1-10mmol/L;
adenosine Triphosphate (ATP) with a concentration of 1-10mmol/L;
l-glutamine synthetase (L-GS) with the concentration of 1-100U/L;
NADH with a concentration of 0.45mmol/L;
lactate Dehydrogenase (LDH) at a concentration of 1-10KU/L;
1% of surfactant, the concentration of which is 0.01% -0.5%;
1 percent of preservative with the concentration of 0.01 percent to 0.5 percent;
reagent R2 consists of:
buffer solution 2, pH6.0-7.0, concentration is 10-100mmol/L;
glutaric acid disodium salt, the concentration is 1-10mmol/L;
adenosine Triphosphate (ATP) with a concentration of 1-10mmol/L;
L-GS with the concentration of 1-100U/L;
GLDH, concentration 10-100KU/L;
enzyme protective agent with concentration of 0.2% -2%;
surfactant 2 with concentration of 0.01-0.5%;
preservative 2 with concentration of 0.01-0.5%.
In the above technical solution, preferably, the buffer solution 1 is tris, carbonate buffer solution (Na) 2 CO 3 /NaHCO 3 ) One or more of AMPSO and glycine/sodium hydroxide.
In the above technical scheme, preferably, the buffer solution 2 is bis-tris, phosphate buffer solution (NaH) 2 PO 4 /Na 2 HPO 4 ) One or more of MES and PIPES.
In the technical scheme, the surfactant 1 is preferably one or more of TritonX-100, tween-20, tween-80 and dodecyl betaine.
In the technical scheme, the surfactant 2 is preferably one or more of brij-35, OP-10, NP-10 and tridecyl polyether-12.
In the above technical solution, it is preferable that the preservative 1 and the preservative 2 are MIT, naN 3 One or more of Chloroacetamide (CAA) and imidazolinyl urea (IZU).
In the above technical solution, preferably, the enzyme protecting agent is one or more of sucrose, trehalose, mannitol, BSA, and HSA.
In the above technical solution, it is preferable that the stable dual reagent blood ammonia assay kit comprises:
the reagent R1 consists of:
tris, pH8.5, concentration 200mmol/L;
alpha-ketoglutaric acid, the concentration is 3mmol/L;
glutaric acid disodium salt, the concentration is 5mmol/L;
ATP with the concentration of 2mmol/L;
L-GS with the concentration of 5U/L;
NADH with a concentration of 0.45mmol/L;
LDH with the concentration of 10KU/L;
TritonX-100 with a concentration of 0.05%;
MIT, concentration 0.1%;
the reagent R2 consists of:
bis-tris, pH6.0, concentration 20mmol/L;
glutaric acid disodium salt, the concentration is 5mmol/L;
ATP with the concentration of 2mmol/L;
L-GS, the concentration is 5U/L;
GLDH, concentration 20KU/L;
mannitol at a concentration of 1%;
brij-35, concentration 0.1%;
NaN 3 the concentration was 0.05%.
In the above technical solution, it is preferable that the stable dual reagent blood ammonia assay kit comprises:
the composition of reagent R1 is as follows:
glycine/sodium hydroxide, concentration pH8.5, 150mmol/L;
alpha-ketoglutaric acid, the concentration is 10mmol/L;
glutaric acid disodium salt, the concentration is 3mmol/L;
ATP with the concentration of 1mmol/L;
L-GS, the concentration is 3U/L;
NADH with a concentration of 0.45mmol/L;
LDH, the concentration is 5KU/L;
tween-20, concentration 0.1%;
CAA, concentration 0.1%;
reagent R2 consists of:
PIPES, pH6.5, concentration of 50mmol/L;
glutaric acid disodium salt, the concentration is 10mmol/L;
ATP with the concentration of 1mmol/L;
L-GS, the concentration is 3U/L;
GLDH, concentration 60KU/L;
BSA, concentration 2%;
OP-10 with the concentration of 0.05 percent;
CAA, concentration 0.1%.
In the above technical solution, it is preferable that the stable dual reagent blood ammonia assay kit comprises:
the composition of reagent R1 is as follows:
AMPSO, pH8.6, concentration of 100mmol/L,
alpha-ketoglutaric acid, the concentration is 5mmol/L;
glutaric acid disodium salt, the concentration is 8mmol/L;
ATP with the concentration of 5mmol/L;
L-GS, the concentration is 6U/L;
NADH with a concentration of 0.45mmol/L;
LDH, the concentration is 1KU/L;
tween-80, concentration 0.02%;
IZU with the concentration of 0.2%;
the reagent R2 consists of:
MES, pH6.0, concentration of 100mmol/L;
glutaric acid disodium salt, the concentration is 8mmol/L;
ATP with the concentration of 5mmol/L;
L-GS with the concentration of 6U/L;
GLDH with a concentration of 30KU/L;
BSA, concentration 2%;
brij-35, concentration 0.02%;
IZU, concentration 0.2%.
The invention has the beneficial effects that:
according to the stable dual-reagent blood ammonia determination kit, the L-glutamine synthetase (L-GS) and the glutaric acid disodium salt are applied to a reaction system, and the stability of the reagent storage process is ensured under the condition that the main reaction is not influenced.
The stable dual-reagent blood ammonia determination kit can eliminate the interference of impurity AMM, has good stability and is based on an enzyme kinetics method. The kit has the advantages of simple preparation process, high accuracy and good repeatability, and is suitable for in vitro determination of the ammonia content in serum and plasma samples.
Drawings
The invention is described in further detail below with reference to the drawings and the detailed description.
FIG. 1 is a zero time calibration graph of the kit of example 2 of the present invention;
FIG. 2 is a graph of a 14-day high temperature calibration of the kit of example 2 of the present invention;
FIG. 3 is a graph of an onboard 35 day calibration of the kit of example 2 of the present invention;
FIG. 4 is a graph showing 13-month calibration of the expiration date of the kit of example 2 of the present invention.
Detailed Description
The invention idea of the invention is as follows: the invention aims to provide a blood ammonia determination kit which can eliminate the interference of impurity AMM, has good stability and is based on an enzyme kinetics method. The kit has the advantages of simple preparation process, high accuracy and good repeatability, and is suitable for in vitro determination of the ammonia content in serum and plasma samples.
The reaction for eliminating the interference of the impurity AMM is as follows:
the double-reagent blood ammonia determination kit (glutamate dehydrogenase method) comprises a reagent R1 and a reagent R2.
R1 consists of:
r2 consists of:
it is preferable that: the buffer solution 1 is tris and carbonate buffer solution (Na) 2 CO 3 /NaHCO 3 ) One or more of AMPSO and glycine/sodium hydroxide; the buffer solution 2 is bis-tris, phosphate buffer solution (NaH) 2 PO 4 /Na 2 HPO 4 ) One or more of MES and PIPES.
It is preferable that: the surfactant 1 is one or more of TritonX-100, tween-20, tween-80 and dodecyl betaine; the surfactant 2 is one or more of brij-35, OP-10, NP-10 and tridecyl polyether-12.
It is preferable that: the preservatives 1 and 2 are MIT and NaN 3 One or more of Chloroacetamide (CAA) and imidazolinyl urea (IZU).
It is preferable that: the enzyme protective agent is one or more of sucrose, trehalose, mannitol, BSA and HSA.
The key point of the invention is to eliminate the interference of impurities and AMM generated in the storage process without affecting the main reaction.
This requires two things to work:
(1) The appropriate L-GS use concentration (1-100U/L) is obtained through experiments. FumihikoYamaguchi et al use L-GS at greater concentrations (1000U/L) and use AMP to inhibit the reaction of L-GS with AMM, adding cost and uncertainty to the reaction; the concentration of L-GS used in the invention can ensure that redundant AMM is removed in time, and the main reaction in the test is not influenced.
(2) No material affecting the main reaction is introduced. Glutamate is also used as a substrate for L-GS:
however, glutamic acid is a product of the main reaction, and if glutamic acid is used as a substrate for L-GS, the sensitivity of the main reaction is greatly reduced, and therefore, the most suitable substrate is glutaric acid disodium salt.
The terms used in the present invention generally have meanings commonly understood by those of ordinary skill in the art, unless otherwise specified. The present invention is described in further detail below with reference to specific examples and with reference to the data. It will be understood that this example is intended to illustrate the invention and not to limit the scope of the invention in any way.
In the following examples, various procedures and methods not described in detail are conventional methods well known in the art. Materials, reagents, devices, instruments, apparatuses and the like used in the following examples are commercially available unless otherwise specified.
Example 1: preparation of blood ammonia determination kit
R1 consists of:
preparation of reagent R1: adding a full batch of purified water into a container, sequentially adding a full batch of tris, alpha-KG, disodium glutarate, ATP, tritonX-100 and MIT into the container in sequence, stirring the mixture until the mixture is completely dissolved, adjusting the pH to 8.5 +/-0.05 (25 +/-0.5 ℃), sequentially adding a full batch of L-GS, NADH and LDH into the mixture in sequence, stirring the mixture until the mixture is completely dissolved, and filtering the mixture by using a 0.22 mu M filter membrane.
R2 consists of:
| name (R) | Concentration of |
| bis-tris，pH6.0 | 20mmol/L |
| Glutaric acid disodium salt | 5mmol/L |
| ATP | 2mmol/L |
| L-GS | 5U/L |
| GLDH | 20KU/L |
| Mannitol | 1％ |
| brij-35 | 0.1％ |
| NaN 3 | 0.05％ |
Preparation of reagent R2: adding a full batch of purified water into a container, and sequentially adding a full batch of bis-tris, glutaric acid disodium salt, ATP, mannitol, brij-35 and NaN into the container in sequence 3 Sequentially stirring until completely dissolved, adjusting pH to 6.0 (25 + -0.5 deg.C), sequentially adding L-GS and GLDH in the same amount, and sequentially stirringAfter stirring to complete dissolution, the mixture was filtered through a 0.22 μ M filter.
Example 2: preparation of blood ammonia determination kit
R1 consists of:
preparation of reagent R1: adding a whole batch of purified water into a container, sequentially adding a whole batch of glycine/sodium hydroxide, alpha-KG, disodium glutarate, ATP, tween-20 and CAA into the container, sequentially stirring until the glycine/sodium hydroxide, the alpha-KG, the disodium glutarate, the ATP, the Tween-20 and the CAA are completely dissolved, adjusting the pH to 8.5 +/-0.05 (25 +/-0.5 ℃), sequentially adding a whole batch of L-GS, NADH and LDH, sequentially stirring until the L-GS, the NADH and the LDH are completely dissolved, and filtering by using a 0.22 mu M filter membrane.
R2 consists of:
| name (R) | Concentration of |
| PIPES，pH6.5 | 50mmol/L |
| Glutaric acid disodium salt | 10mmol/L |
| ATP | 1mmol/L |
| L-GS | 3U/L |
| GLDH | 60KU/L |
| BSA | 2％ |
| OP-10 | 0.05％ |
| CAA | 0.1％ |
Preparation of reagent R2: adding a full batch of purified water into a container, sequentially adding a full batch of PIPES, disodium glutarate, ATP, BSA, OP-10 and CAA into the container in sequence, stirring until the mixture is completely dissolved, adjusting the pH to 6.5 (25 +/-0.5 ℃), sequentially adding a full batch of L-GS and GLDH into the container in sequence, stirring until the mixture is completely dissolved, and filtering with a 0.22 mu M filter membrane.
Example 3: preparation of blood ammonia determination kit
R1 consists of:
| name(s) | Concentration of |
| AMPSO，pH8.6 | 100mmol/L |
| Alpha-ketoglutaric acid | 5mmol/L |
| Glutaric acid disodium salt | 8mmol/L |
| ATP | 5mmol/L |
| L-GS | 6U/L |
| NADH | 0.45mmol/L |
| LDH | 1KU/L |
| Tween-80 | 0.02％ |
| IZU | 0.2％ |
Preparation of reagent R1: adding a whole batch of purified water into a container, sequentially adding a whole batch of AMPSO, alpha-KG, disodium glutarate, ATP, tween-80 and IZU into the container in sequence, stirring the mixture until the mixture is completely dissolved, adjusting the pH to 8.6 +/-0.05 (25 +/-0.5 ℃), sequentially adding a whole batch of L-GS, NADH and LDH into the mixture in sequence, stirring the mixture until the mixture is completely dissolved, and filtering the mixture by using a 0.22 mu M filter membrane.
R2 consists of:
| name(s) | Concentration of |
| MES，pH6.0 | 100mmol/L |
| Glutaric acid disodium salt | 8mmol/L |
| ATP | 5mmol/L |
| L-GS | 6U/L |
| GLDH | 30KU/L |
| BSA | 2％ |
| brij-35 | 0.02％ |
| IZU | 0.2％ |
Preparation of reagent R2: adding a full batch of purified water into a container, sequentially adding a full batch of MES, disodium glutarate, ATP, BSA, brij-35 and IZU into the container in sequence, stirring the mixture until the mixture is completely dissolved, adjusting the pH to 6.0 (25 +/-0.5 ℃), sequentially adding a full batch of L-GS and GLDH into the mixture in sequence, stirring the mixture until the mixture is completely dissolved, and filtering the mixture by using a 0.22 mu M filter membrane.
Example 4: performance inspection
1. Evaluation of high-temperature accelerated stability
The reagents of examples 1 to 3 were placed in a 37 ℃ incubator and a 2-8 ℃ refrigerator, and the reagents were taken out on days 7 and 14 while testing the quality control low/high values as shown in table 1. The test results were compared with 2-8 ℃ cold storage reagent and the relative deviation (%) was calculated and required to be less than 15%.
TABLE 1 high temperature stability test results
As can be seen from the results in Table 1, the reagents of examples 1 to 3 according to the invention were stable at 37 ℃ for 14 days, with example 2 having the best stability and all being better than the comparative reagent (Shaoxing Shengkang: ammonia assay kit, batch No. 200113K 36).
2. Evaluation of on-board stability
After the reagents of examples 1 to 3 were decapped, they were placed in a fully automatic biochemical analyzer, and the decapped reagents and 2-8 ℃ cold storage reagents were tested for quality control low/high values at the same time on days 7, 14, 21, 30, and 35, as shown in table 2. The test results are compared with the refrigerated reagent at the temperature of 2-8 ℃, and the relative deviation (%) is calculated and is required to be less than 15%.
TABLE 2 airborne stability test results
As can be seen from the results in Table 2, the reagents of examples 1 to 3 of the present invention were stable for 30 days under the decap condition, and the reagent of example 2 was the most stable.
3. Evaluation of shelf-Life stability
The evaluation of stability during expiration was performed using the reagent prepared in example 2, and the dispensed reagents were stored in an environment of 2 to 8 ℃, and the reagents were tested every 3 months for 13 months in succession, as shown in table 3 and fig. 1 to 4. The test results were compared to the zero time reagent test results and the relative deviation (%) calculated, requiring less than 15%.
TABLE 3 Effect stability test results
As can be seen from the results in Table 3, the reagent of example 2 of the present invention was stable for at least 12 months without change in performance.
4. Repeatability survey
The low/high value of the quality control material is determined by using the same batch number reagent in example 2, and the test is repeated for 20 times, and the result is shown in table 4.
TABLE 4 results of reproducibility measurement
As can be seen from Table 4, example 2 has a repeatability index CV of no more than 4.0%.
The above description is only of the preferred embodiments of the present invention, and it should be noted that: it will be appreciated by those skilled in the art that the present invention is not limited by the embodiments illustrated above, which are merely illustrative of the principles and preferred embodiments of the invention, but that various modifications and adaptations of the invention are possible within the scope of the invention as claimed.
Claims (10)
1. A stable double-reagent blood ammonia determination reagent kit is characterized by comprising a reagent R1 and a reagent R2;
the composition of reagent R1 is as follows:
buffer solution 1, pH8.0-9.0, concentration 50-200mmol/L;
alpha-ketoglutaric acid, the concentration is 1-10mmol/L;
glutaric acid disodium salt, the concentration is 1-10mmol/L;
ATP with the concentration of 1-10mmol/L;
L-GS with the concentration of 1-100U/L;
NADH with a concentration of 0.45mmol/L;
LDH, the concentration is 1-10KU/L;
1% of surfactant, the concentration of which is 0.01% -0.5%;
preservative 1, the concentration is 0.01% -0.5%;
the reagent R2 consists of:
buffer solution 2, pH6.0-7.0, concentration 10-100mmol/L;
glutaric acid disodium salt, the concentration is 1-10mmol/L;
ATP with concentration of 1-10mmol/L;
L-GS with the concentration of 1-100U/L;
GLDH with the concentration of 10-100KU/L;
enzyme protective agent with concentration of 0.2% -2%;
surfactant 2 with concentration of 0.01-0.5%;
preservative 2 with concentration of 0.01-0.5%.
2. The stable dual reagent blood ammonia assay kit of claim 1, wherein said buffer 1 is one of tris, carbonate buffer, AMPSO and glycine/sodium hydroxide.
3. The stable dual reagent blood ammonia assay kit of claim 1, wherein buffer 2 is one of bis-tris, phosphate buffered saline, MES and PIPES.
4. The stable dual reagent ammonia blood assay kit of claim 1, wherein surfactant 1 is one of triton x-100, tween-20, tween-80 and dodecyl betaine.
5. The stable two-reagent blood ammonia assay kit according to claim 1, wherein said surfactant 2 is one of brij-35, OP-10, NP-10, and trideceth-12.
6. The stable dual reagent blood ammonia assay kit of claim 1, wherein said preservative 1 and preservative 2 are MIT, naN 3 One of chloroacetamide and imidazolinyl urea.
7. The stable dual reagent kit for assaying blood ammonia of claim 1, wherein the enzyme protecting agent is one of sucrose, trehalose, mannitol, BSA and HSA.
8. The stable dual reagent blood ammonia assay kit of claim 1, consisting of:
the composition of reagent R1 is as follows:
tris, pH8.5, concentration 200mmol/L;
alpha-ketoglutaric acid, the concentration is 3mmol/L;
glutaric acid disodium salt, the concentration is 5mmol/L;
ATP with the concentration of 2mmol/L;
L-GS, the concentration is 5U/L;
NADH with a concentration of 0.45mmol/L;
LDH, the concentration is 10KU/L;
TritonX-100 with concentration of 0.05%;
MIT, concentration 0.1%;
the reagent R2 consists of:
bis-tris, at a concentration of pH6.0, 20mmol/L;
glutaric acid disodium salt, the concentration is 5mmol/L;
ATP with the concentration of 2mmol/L;
L-GS, the concentration is 5U/L;
GLDH with a concentration of 20KU/L;
mannitol at a concentration of 1%;
brij-35, concentration 0.1%;
NaN 3 the concentration was 0.05%.
9. The stable dual reagent blood ammonia assay kit of claim 1, consisting of:
the reagent R1 consists of:
glycine/sodium hydroxide, pH8.5, concentration of 150mmol/L;
alpha-ketoglutaric acid, the concentration is 10mmol/L;
glutaric acid disodium salt, the concentration is 3mmol/L;
ATP with the concentration of 1mmol/L;
L-GS with the concentration of 3U/L;
NADH with a concentration of 0.45mmol/L;
LDH, the concentration is 5KU/L;
tween-20, concentration 0.1%;
CAA, concentration 0.1%;
reagent R2 consists of:
PIPES, pH6.5, concentration 50mmol/L;
glutaric acid disodium salt, the concentration is 10mmol/L;
ATP with the concentration of 1mmol/L;
L-GS, the concentration is 3U/L;
GLDH, concentration 60KU/L;
BSA, concentration 2%;
OP-10 with concentration of 0.05%;
CAA, concentration 0.1%.
10. The stable dual reagent blood ammonia assay kit of claim 1, consisting of:
the reagent R1 consists of:
AMPSO, pH8.6, concentration 100mmol/L,
alpha-ketoglutaric acid, the concentration is 5mmol/L;
glutaric acid disodium salt, the concentration is 8mmol/L;
ATP with the concentration of 5mmol/L;
L-GS, the concentration is 6U/L;
NADH with a concentration of 0.45mmol/L;
LDH with the concentration of 1KU/L;
tween-80, concentration 0.02%;
IZU with the concentration of 0.2%;
reagent R2 consists of:
MES, pH6.0, concentration 100mmol/L;
glutaric acid disodium salt, the concentration is 8mmol/L;
ATP with the concentration of 5mmol/L;
L-GS, the concentration is 6U/L;
GLDH concentration 30KU/L;
BSA, concentration 2%;
brij-35, concentration 0.02%;
IZU, concentration 0.2%.
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| CN202110332146.6A CN113075139B (en) | 2021-03-29 | 2021-03-29 | Stable double-reagent blood ammonia determination kit |
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| CN113075139B true CN113075139B (en) | 2022-10-11 |
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| CN202110332146.6A Active CN113075139B (en) | 2021-03-29 | 2021-03-29 | Stable double-reagent blood ammonia determination kit |
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Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS623800A (en) * | 1985-06-27 | 1987-01-09 | Takara Shuzo Co Ltd | Determination of ammonia or atp |
| JPH05219991A (en) * | 1992-02-10 | 1993-08-31 | Taunzu:Kk | Method for determination using enzyme reaction |
| CN1778937A (en) * | 2004-11-23 | 2006-05-31 | 苏州艾杰生物科技有限公司 | Determination of blood ammonia content and blood ammonia diagnostic reagent kit |
| CN101403691A (en) * | 2008-11-14 | 2009-04-08 | 苏州艾杰生物科技有限公司 | Melamine or tricyanic acid measuring method and its reagent kit or test paper strip |
| CN103293323A (en) * | 2013-06-14 | 2013-09-11 | 南京医科大学第二附属医院 | Luteinizing hormone nano-magnetic particle chemiluminescence quantitative immunoassay kit and preparation method thereof |
| CN104483280A (en) * | 2014-12-23 | 2015-04-01 | 阜阳师范学院 | Method for rapidly detecting ammonia nitrogen removal rate |
| CN104561237A (en) * | 2013-10-21 | 2015-04-29 | 大连市沙河口区中小微企业服务中心 | Blood ammonia kit |
| CN107014969A (en) * | 2016-01-27 | 2017-08-04 | 北京凯茂正奇科技有限公司 | Detect the agent prescription of urea and ammonia content |
| CN107688017A (en) * | 2016-08-05 | 2018-02-13 | 爱科来株式会社 | Quantitative approach, quantitative reagent, quantification kit, test piece and the ammonia proportioning device of ammonia |
| CN107974485A (en) * | 2016-10-25 | 2018-05-01 | 爱科来株式会社 | Ammonia quantitative approach, quantification kit, testing piece and ammonia proportioning device |
| CN110308282A (en) * | 2019-06-21 | 2019-10-08 | 中生北控生物科技股份有限公司 | A kind of stable homocysteine Enzymatic cycling detection kit |
| CN110441522A (en) * | 2019-07-29 | 2019-11-12 | 迪瑞医疗科技股份有限公司 | Superoxide dismutase assay kit in a kind of serum |
| CN111778312A (en) * | 2020-06-15 | 2020-10-16 | 迪瑞医疗科技股份有限公司 | Kit for determining glycylproline dipeptide aminopeptidase in serum |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1053548A (en) * | 1974-04-29 | 1979-05-01 | Charles Allain | Method and reagent for reducing errors in enzymatic assays |
| JP2000232898A (en) * | 1999-02-17 | 2000-08-29 | Kyowa Medex Co Ltd | Quantitative analysis of substance and reagent therefor |
| CN101169411A (en) * | 2006-10-24 | 2008-04-30 | 苏州艾杰生物科技有限公司 | Ammonia diagnosis/determination reagent kit and ammonia concentration determination method |
| CN101464346A (en) * | 2007-12-19 | 2009-06-24 | 苏州艾杰生物科技有限公司 | Ammonia ion diagnosis/measuring reagent kit and ammonia ion concentration determination method |
| CN101464377A (en) * | 2007-12-19 | 2009-06-24 | 苏州艾杰生物科技有限公司 | Ammonia ion diagnosis/measuring reagent kit and ammonia ion concentration determination method |
| CN104198473B (en) * | 2014-08-14 | 2017-07-07 | 上海睿康生物科技有限公司 | A kind of uric acid detection kit of stabilization |
| CN111727975A (en) * | 2020-07-03 | 2020-10-02 | 山东中新科农生物科技有限公司 | Weeding composition, compound herbicide and application |
-
2021
- 2021-03-29 CN CN202110332146.6A patent/CN113075139B/en active Active
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS623800A (en) * | 1985-06-27 | 1987-01-09 | Takara Shuzo Co Ltd | Determination of ammonia or atp |
| JPH05219991A (en) * | 1992-02-10 | 1993-08-31 | Taunzu:Kk | Method for determination using enzyme reaction |
| CN1778937A (en) * | 2004-11-23 | 2006-05-31 | 苏州艾杰生物科技有限公司 | Determination of blood ammonia content and blood ammonia diagnostic reagent kit |
| CN101403691A (en) * | 2008-11-14 | 2009-04-08 | 苏州艾杰生物科技有限公司 | Melamine or tricyanic acid measuring method and its reagent kit or test paper strip |
| CN103293323A (en) * | 2013-06-14 | 2013-09-11 | 南京医科大学第二附属医院 | Luteinizing hormone nano-magnetic particle chemiluminescence quantitative immunoassay kit and preparation method thereof |
| CN104561237A (en) * | 2013-10-21 | 2015-04-29 | 大连市沙河口区中小微企业服务中心 | Blood ammonia kit |
| CN104483280A (en) * | 2014-12-23 | 2015-04-01 | 阜阳师范学院 | Method for rapidly detecting ammonia nitrogen removal rate |
| CN107014969A (en) * | 2016-01-27 | 2017-08-04 | 北京凯茂正奇科技有限公司 | Detect the agent prescription of urea and ammonia content |
| CN107688017A (en) * | 2016-08-05 | 2018-02-13 | 爱科来株式会社 | Quantitative approach, quantitative reagent, quantification kit, test piece and the ammonia proportioning device of ammonia |
| CN107974485A (en) * | 2016-10-25 | 2018-05-01 | 爱科来株式会社 | Ammonia quantitative approach, quantification kit, testing piece and ammonia proportioning device |
| CN110308282A (en) * | 2019-06-21 | 2019-10-08 | 中生北控生物科技股份有限公司 | A kind of stable homocysteine Enzymatic cycling detection kit |
| CN110441522A (en) * | 2019-07-29 | 2019-11-12 | 迪瑞医疗科技股份有限公司 | Superoxide dismutase assay kit in a kind of serum |
| CN111778312A (en) * | 2020-06-15 | 2020-10-16 | 迪瑞医疗科技股份有限公司 | Kit for determining glycylproline dipeptide aminopeptidase in serum |
Non-Patent Citations (7)
| Title |
|---|
| alpha-Aminomethylglutarate, a beta-amino analog of glutamate that interacts with glutamine synthetase and the enzymes that catalyze glutathione synthesis;R Sekura et al.;《Journal of Biological Chemistry》;19760425;第251卷(第8期);第2263-2270页 * |
| α-酮戊二酸对浑球红假单胞菌谷氨酰胺合成酶的变构调节;王星 等;《全国首届青年植物生理工作者学术讨论会》;19880603;第18页 * |
| 毛细管电泳快速测定脑脊液中谷氨酰胺含量;刘阳等;《临床检验杂志》;20021230;第20卷(第06期);第352-353页 * |
| 血氨检测在临床肝脏疾病中的应用价值;李晓光 等;《国际检验医学杂志》;20120724;第33卷(第11期);第1340-1342页 * |
| 血清谷氨酸脱氢酶对肝病诊断的临床评价;李漫松 等;《吉林医学》;19970131(第1期);第2页 * |
| 血清钾的色氨酸酶法测定;李永军;《国际检验医学杂志》;19930525;第14卷(第03期);第135页 * |
| 食品塑料包装溶出酞酸酯DBP与DEHP的测定;王秀娟 等;《食品研究与开发》;20150815;第36卷(第15期);第95-98页 * |
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