CN111778312A - Kit for determining glycylproline dipeptide aminopeptidase in serum - Google Patents
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Abstract
A kit for determining glycylproline dipeptidyl aminopeptidase in serum belongs to the technical field of kits. The kit of the invention comprises an R1 reagent and an R2 reagent; the R1 reagent is a buffer solution containing a protective agent and a preservative; the buffer solution is 3- [ (1, 1-dimethyl-2-hydroxyethyl) amino ] -2-hydroxypropanesulfonic acid buffer solution, triethanolamine buffer solution or trihydroxyaminomethane-hydrochloric acid buffer solution; the protective agent is diglycine; the preservative is Proclin 300; the R2 reagent is a buffer solution containing a surfactant, a preservative and a substrate; the buffer solution is tartaric acid buffer solution; the surfactant is one or two of polyoxyethylene ether and fatty alcohol polyoxyethylene ether of alkylphenol; the preservative is Proclin 300; the substrate is glycylprolyl p-nitroaniline. The reagent kit has high unsealing stability and high-temperature stability of the reagent, and good repeatability; false results of clinical tests are avoided, and the detection efficiency and accuracy are high.
Description
Technical Field
The invention belongs to the technical field of kits, and particularly relates to a kit for determining glycylproline dipeptidyl aminopeptidase in serum.
Background
From commercial enzyme product Acylase I, Hopsu-Havu and Glenner, glycylproline dipeptide aminopeptidase (GPDA) was first discovered and isolated in 1966, and then GPDA was confirmed in tissues such as rat liver, rat kidney, pig kidney, human and various animal sera, human salivary gland and saliva, bovine and human dental pulp and gum.
GPDA is mainly found in the salivary gland, submandibular gland and other tissues of the human body, and saliva and blood also contain the enzyme. The main role of GPDA is to hydrolyze the peptide chain between glycylproline and other amino acids located at the N-terminus of the peptide chain. The enzyme activity in blood serum of liver disease patients is increased, and gastric cancer patients are decreased. The GPDA activity in the serum of patients with primary liver cancer (PHC) and secondary liver cancer is obviously higher than that in patients with chronic hepatitis, liver cirrhosis, cholelithiasis, obstructive jaundice and normal control group. Acute hepatitis, chronic live liver, liver cirrhosis, obstructive jaundice, etc., and GPDA in blood serum may be increased to different extent, which is not as high as that of liver cancer patients. However, in severe hepatitis and alcoholic hepatitis, GPDA in blood serum is higher than that in liver cancer patients. The GPDA of the serum of a patient with gastric cancer is obviously reduced, and is generally about 1/2 of a normal person. Other benign gastrointestinal lesions. The GPDA also decreased slightly. The gastric ulcer, which is the major descending factor, is chronic gastritis and duodenal bulbar ulcer. After gastric cancer resection, GPDA in the serum of a patient tends to rise.
In the prior art, a substrate required for determining the glycylproline dipeptide aminopeptidase is glycylproline p-nitroaniline p-toluenesulfonate, and the glycylproline p-nitroaniline p-toluenesulfonate is unstable after being directly prepared into an aqueous solution and is usually used on the same day, so that the inconvenience, waste and cost increase are caused for clinical biochemical tests. Therefore, it is important to improve the stability of the reagent.
Tartaric acid (tartaric), a carboxylic acid, is present in a variety of plants, such as grapes and tamarind, and is also one of the major organic acids in wine. The antioxidant can be added into food to impart sour taste to the food. This use is similar to citric acid. Tartaric acid is used together with tannin as mordant of acid dye and also used for some developing and fixing operations in photographic industry, and iron salt thereof has photosensitivity and thus can be used for making blueprints. Tartaric acid can be complexed with various metal ions and can be used as a cleaning agent and a polishing agent for metal surfaces. Tartaric acid is an important auxiliary agent and reducing agent in the mirror making industry, and can control the formation speed of silver mirror and obtain a very uniform coating. However, in the prior art, tartaric acid is not applied to the field of reagents and the application of improving the stability of the reagents is not provided.
Disclosure of Invention
The invention aims to solve the technical problem of instability of reagents used in the detection method of the glycylproline dipeptide aminopeptidase in the prior art, and provides a kit for determining the glycylproline dipeptide aminopeptidase in serum.
The invention provides a kit for determining glycylproline dipeptide aminopeptidase in serum,
comprising an R1 reagent and an R2 reagent;
the R1 reagent is a buffer solution containing a protective agent and a preservative; the buffer solution is 3- [ (1, 1-dimethyl-2-hydroxyethyl) amino ] -2-hydroxypropanesulfonic Acid (AMPSO) buffer solution with the concentration range of 50-100mM, triethanolamine buffer solution with the concentration range of 8-10g/L or trihydroxyaminomethane-hydrochloric acid (Tris-HCl) buffer solution with the concentration range of 60-100 mM; the protective agent is diglycine, and the concentration of the protective agent in the R1 reagent is 5-50 mM; the preservative is Proclin300, and the concentration of the preservative in the R1 reagent is 0.05 wt% -0.25 wt%;
the R2 reagent is a buffer solution containing a surfactant, a preservative and a substrate; the buffer solution is tartaric acid buffer solution with the concentration range of 20-100 mM; the surfactant is one or two of polyoxyethylene ether and fatty alcohol polyoxyethylene ether of alkylphenol, and the concentration of the surfactant in the R2 reagent is 0.1-3 wt%; the preservative is Proclin300, and the concentration of the preservative in the R2 reagent is 0.05 wt% -0.25 wt%; the substrate is glycylprolyl p-nitrobenzene, and the concentration of the substrate in the R2 reagent is 6 mM.
Preferably, in the R1 reagent, the buffer is 3- [ (1, 1-dimethyl-2-hydroxyethyl) amino ] -2-hydroxypropanesulfonic acid buffer with a concentration of 80 mM.
Preferably, the concentration of the protective agent in the R1 reagent is 20 mM.
Preferably, the concentration of the preservative in the R1 reagent is 0.1 wt%.
Preferably, the concentration of the tartaric acid buffer in the R2 reagent is 50 mM.
Preferably, in the R2 reagent, the surfactant is polyoxyethylene lauryl ether, and the concentration is 2 wt%.
Preferably, the concentration of the preservative in the R2 reagent is 0.1 wt%.
Compared with the prior art, the invention has the beneficial effects that:
according to the kit for determining glycylproline dipeptide aminopeptidase in serum, tartaric acid is used, so that the unsealing stability and the high-temperature stability of a reagent are improved, and the detection repeatability is good; the method effectively avoids the false result of clinical test, greatly improves the detection efficiency and accuracy, is suitable for various full-automatic biochemical analyzers, has wider universality and is convenient to popularize.
The kit for determining glycylproline dipeptidyl aminopeptidase in serum has lower raw material cost, can replace imported reagents, and saves the expense of patients.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings used in the detailed description will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a calibration curve of the kit for assaying glycylproline dipeptidyl aminopeptidase in serum provided in example 1 of the present invention.
FIG. 2 is a method of using the kit for glycylproline dipeptidyl aminopeptidase in serum according to the present invention.
Detailed Description
For a further understanding of the invention, preferred embodiments of the invention are described below in conjunction with the detailed description, but it is to be understood that the description is intended to further illustrate the features and advantages of the invention and not to limit the claims to the invention.
The kit for determining glycylproline dipeptidyl aminopeptidase in serum comprises an R1 reagent and an R2 reagent.
Wherein, the R1 reagent is a buffer solution containing a protective agent and a preservative; the buffer solution is 3- [ (1, 1-dimethyl-2-hydroxyethyl) amino ] -2-hydroxypropanesulfonic acid buffer solution with the concentration range of 50-100mM, triethanolamine buffer solution with the concentration range of 8-10g/L or trihydroxyaminomethane-hydrochloric acid buffer solution with the concentration range of 60-100 mM; the protective agent is diglycine, and the concentration of the protective agent in the R1 reagent is 5-50 mM; the preservative is Proclin300, and the concentration of the preservative in the R1 reagent is 0.05 wt% -0.25 wt%;
the R2 reagent is a buffer solution containing a surfactant, a preservative and a substrate; the buffer solution is tartaric acid buffer solution with the concentration range of 20-100 mM; the surfactant is one or two of polyoxyethylene ether and fatty alcohol polyoxyethylene ether of alkylphenol, and the concentration of the surfactant in the R2 reagent is 0.1-3 wt%; the preservative is Proclin300, and the concentration of the preservative in the R2 reagent is 0.05 wt% -0.25 wt%; the substrate is glycylprolyl p-nitrobenzene, and the concentration of the substrate in the R2 reagent is 6 mM.
In the above technical means, the buffer solution in the R1 reagent is preferably 3- [ (1, 1-dimethyl-2-hydroxyethyl) amino ] -2-hydroxypropanesulfonic acid buffer solution with a concentration of 80 mM.
In the above embodiment, the concentration of the protecting agent in the R1 reagent is preferably 20 mM.
In the above embodiment, the concentration of the preservative in the R1 reagent is preferably 0.1 wt%.
In the above embodiment, the concentration of the tartaric acid buffer in the R2 reagent is preferably 50 mM.
In the technical scheme, in the R2 reagent, the surfactant is preferably polyoxyethylene lauryl ether, and the concentration is preferably 2 wt%.
In the above embodiment, the concentration of the preservative in the R2 reagent is preferably 0.1 wt%.
The detection principle of the kit for determining glycylproline dipeptidyl aminopeptidase in serum provided by the invention is as follows: under alkaline conditions, GPDA catalyzes the substrate glycylprolyl-p-nitroaniline to be hydrolyzed to generate glycylproline and yellow p-nitroaniline, the latter can cause the increase of absorbance at specific wavelength, and the increase rate of the absorbance is in direct proportion to the activity of GPDA.
The method of using the kit for assaying glycylproline dipeptidyl aminopeptidase in serum according to the present invention is shown in FIG. 2 and Table 1.
TABLE 1
The terms used in the present invention generally have meanings commonly understood by those of ordinary skill in the art, unless otherwise specified.
In order to make the technical solutions of the present invention better understood by those skilled in the art, the present invention will be further described in detail with reference to the following embodiments and the accompanying drawings.
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
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| AMP | 50mM |
| double-glycylglycine | 20mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 50mM |
| Polyoxyethylene lauryl ether | 1.5wt％ |
| Proclin 300 | 0.1wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 2
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
the formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 80mM |
| Polyoxyethylene lauryl ether | 3.0wt％ |
| Proclin 300 | 0.1wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 3
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| triethanolamine | 8g/L |
| Double-glycylglycine | 20mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 30mM |
| Polyoxyethylene lauryl ether | 2.0wt％ |
| Proclin 300 | 0.1wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 4
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| Tris-HCl | 60mM |
| double-glycylglycine | 5mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 20mM |
| Polyoxyethylene lauryl ether | 0.5wt％ |
| Proclin 300 | 0.1wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 5
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| Tris-HCl | 80 mM |
| double-glycylglycine | 5mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 50mM |
| Polyoxyethylene lauryl ether | 0.5wt％ |
| Proclin 300 | 0.1wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 6
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| Tris-HCl | 100mM |
| double-glycylglycine | 5mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 100mM |
| Polyoxyethylene lauryl ether | 0.5wt％ |
| Proclin 300 | 0.25wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 7
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| Tris-HCl | 80mM |
| double-glycylglycine | 20mM |
| Proclin 300 | 0.05wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 50mM |
| Polyoxyethylene lauryl ether | 0.5wt％ |
| Proclin 300 | 0.05wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 8
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| Tris-HCl | 80mM |
| double-glycylglycine | 20mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
example 9
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| AMP | 80mM |
| double-glycylglycine | 20mM |
| Proclin 300 | 0.25wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 50mM |
| Polyoxyethylene lauryl ether | 0.5wt％ |
| Proclin 300 | 0.25wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
Example 10
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| triethanolamine | 10g/L |
| Double-glycylglycine | 20mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
example 11
Glycylproline dipeptide aminopeptidase assay kit:
the formula of the R1 reagent is (the solvent is purified water):
| Tris-HCl | 80mM |
| double-glycylglycine | 20mM |
| Proclin 300 | 0.1wt％ |
| pH | 7.8±0.05(25±0.5℃) |
The formula of the R2 reagent is (the solvent is purified water):
| tartaric acid | 50mM |
| Polyoxyethylene lauryl ether | 2wt％ |
| Proclin 300 | 0.1wt％ |
| pH | 4.5±0.05(25±0.5℃) |
| Glycylprolyl-p-nitroaniline | 6mM |
The performance test was performed on the kit for assaying glycylproline dipeptidyl aminopeptidase in serum of example 1 to example 11.
1. Accuracy survey
The high/low value quality control materials are respectively tested by using the kit for measuring glycylproline dipeptidyl aminopeptidase in serum of example 1 to example 6, the test is repeated for 3 times, the average value is taken, the result average value is compared with the theoretical value of the high/low value quality control materials, the relative deviation (%) is calculated, and the relative deviation is required to be less than 10%.
TABLE 1 accuracy test results
As can be seen from the results in Table 1, the results of the high/low value quality control test using the kit for assaying glycylproline dipeptidyl aminopeptidase in serum according to examples 1 to 6 of the present invention all meet the standard requirements, wherein the relative deviation of example 5 is the smallest, and the accuracy test result is the best.
2. Stability survey
High-temperature stability: the R1 reagent and the R2 reagent of examples 7 to 9 were placed in a 37 ℃ incubator and a 2 to 8 ℃ refrigerator, respectively, and the reagents were taken out on days 7, 14 and 21, respectively, and were simultaneously tested. And respectively testing the high/low value quality control products, repeating the test for 3 times, taking an average value, comparing the average value of the high-temperature test result with the average value of the test result at 2-8 ℃, and calculating the relative deviation (%), wherein the relative deviation is required to be less than 15%.
Note: after the R1 reagent and the R2 reagent at 37 ℃ were taken out from the incubator, the R1 reagent and the R2 reagent, which were refrigerated at 2 to 8 ℃, were placed to the same temperature and then tested.
TABLE 2 high temperature stability test results
Airborne stability: the R1 reagent and the R2 reagent of examples 10 and 11 were decapsulated, placed in a fully automatic biochemical analyzer, tested for high/low quality control, and the decapsulated R1 reagent and R2 reagent were removed on days 7, 14, 21, and 28, and tested simultaneously with the R1 reagent and R2 reagent refrigerated at 2-8 ℃. And repeating the test for 3 times, taking an average value, comparing the average value of the high-temperature test result with the average value of the cold storage test result at the temperature of 2-8 ℃, and calculating the relative deviation (%), wherein the relative deviation is required to be less than 15%.
TABLE 3 airborne stability test results
As can be seen from the results of tables 2 and 3, the R1 reagent and the R2 reagent of examples 7 to 9 of the present invention were stable for 21 days at a high temperature of 37 ℃ and the R1 reagent and the R2 reagent of example 8 were most stable at a high temperature of 37 ℃; the R1 reagent and the R2 reagent of examples 10 to 11 of the present invention were stable for 28 days under the unsealed condition, and the stability of example 11 was the best.
3. Calibration curve
The nine-strength calibrator was assayed using the kit for determining glycylproline dipeptidyl aminopeptidase in serum of example 1, and a calibration curve was drawn with the calibrator concentration as the abscissa and the absorbance value as the ordinate, as shown in fig. 1.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications of the present invention will be apparent to those of ordinary skill in the art in light of the above teachings. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications of the invention may be made without departing from the spirit or scope of the invention.
Claims (7)
1. A kit for determining glycylproline dipeptidyl aminopeptidase in serum is characterized in that,
comprising an R1 reagent and an R2 reagent;
the R1 reagent is a buffer solution containing a protective agent and a preservative; the buffer solution is 3- [ (1, 1-dimethyl-2-hydroxyethyl) amino ] -2-hydroxypropanesulfonic acid buffer solution with the concentration range of 50-100mM, triethanolamine buffer solution with the concentration range of 8-10g/L or trihydroxyaminomethane-hydrochloric acid buffer solution with the concentration range of 60-100 mM; the protective agent is diglycine, and the concentration of the protective agent in the R1 reagent is 5-50 mM; the preservative is Proclin300, and the concentration of the preservative in the R1 reagent is 0.05 wt% -0.25 wt%;
the R2 reagent is a buffer solution containing a surfactant, a preservative and a substrate; the buffer solution is tartaric acid buffer solution with the concentration range of 20-100 mM; the surfactant is one or two of polyoxyethylene ether and fatty alcohol polyoxyethylene ether of alkylphenol, and the concentration of the surfactant in the R2 reagent is 0.1-3 wt%; the preservative is Proclin300, and the concentration of the preservative in the R2 reagent is 0.05 wt% -0.25 wt%; the substrate is glycylprolyl p-nitrobenzene, and the concentration of the substrate in the R2 reagent is 6 mM.
2. The kit for the determination of glycylproline dipeptidylaminopeptidase in serum as claimed in claim 1, wherein the buffer in the reagent R1 is 3- [ (1, 1-dimethyl-2-hydroxyethyl) amino ] -2-hydroxypropanesulfonic acid buffer at a concentration of 80 mM.
3. The kit for assaying glycylproline dipeptidyl aminopeptidase in serum according to claim 1, wherein the concentration of the protective agent in the R1 reagent is 20 mM.
4. The kit for assaying glycylproline dipeptidyl aminopeptidase in serum according to claim 1, wherein the concentration of the preservative in the reagent R1 is 0.1 wt%.
5. The kit for assaying glycylproline dipeptide aminopeptidase in serum according to claim 1, wherein the concentration of the tartaric acid buffer in the reagent R2 is 50 mM.
6. The kit for assaying glycylproline dipeptide aminopeptidase in serum as claimed in claim 1, wherein in the reagent R2, the surfactant is laureth alcohol at a concentration of 2 wt%.
7. The kit for assaying glycylproline dipeptidyl aminopeptidase in serum according to claim 1, wherein the concentration of the preservative in the reagent R2 is 0.1 wt%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113075139A (en) * | 2021-03-29 | 2021-07-06 | 迪瑞医疗科技股份有限公司 | Stable double-reagent blood ammonia determination kit |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104880423A (en) * | 2015-05-08 | 2015-09-02 | 浙江蓝森生物科技有限公司 | Method, reagent and kit for quantitatively determining activity of glycylproline dipeptidyl aminopeptidase in human serum |
| CN109580505A (en) * | 2018-11-07 | 2019-04-05 | 北京九强生物技术股份有限公司 | A kind of stabiliser compositions |
| CN111154834A (en) * | 2019-12-15 | 2020-05-15 | 金华市强盛生物科技有限公司 | Leucine aminopeptidase detection kit and preparation method thereof |
-
2020
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104880423A (en) * | 2015-05-08 | 2015-09-02 | 浙江蓝森生物科技有限公司 | Method, reagent and kit for quantitatively determining activity of glycylproline dipeptidyl aminopeptidase in human serum |
| CN109580505A (en) * | 2018-11-07 | 2019-04-05 | 北京九强生物技术股份有限公司 | A kind of stabiliser compositions |
| CN111154834A (en) * | 2019-12-15 | 2020-05-15 | 金华市强盛生物科技有限公司 | Leucine aminopeptidase detection kit and preparation method thereof |
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| CN113075139A (en) * | 2021-03-29 | 2021-07-06 | 迪瑞医疗科技股份有限公司 | Stable double-reagent blood ammonia determination kit |
| CN113075139B (en) * | 2021-03-29 | 2022-10-11 | 迪瑞医疗科技股份有限公司 | Stable double-reagent blood ammonia determination kit |
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