CN104215631A - Buffer for determining human serum glutamyltransferase - Google Patents
Buffer for determining human serum glutamyltransferase Download PDFInfo
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- CN104215631A CN104215631A CN201410398200.7A CN201410398200A CN104215631A CN 104215631 A CN104215631 A CN 104215631A CN 201410398200 A CN201410398200 A CN 201410398200A CN 104215631 A CN104215631 A CN 104215631A
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Abstract
The invention discloses a buffer for determining human serum glutamyltransferase. The buffer is used in a kit for determining the human serum glutamyltransferase with trihydroxymethyl aminomethane (Tris)-citric acid as a buffer system, the kit comprises a first reagent and a second reagent, the first reagent is composed of the Tris, glycylglycine, mycose and citric acid, and the second reagent is composed of the Tris, L-glutamic acid gamma-(3-carboxy-4-nitroanilide)ammonium (GPNA), citric acid and sodium azide. The buffer for determining the human serum glutamyltransferase prepared in above mode adopts Tris-citric acid as the buffer system, and the kit matched with the buffer can reach the level of reported commonly-used buffers, has good data determination accuracy and stability, and is worth popularization use.
Description
Technical field
The present invention relates to field of medical examination, particularly relate to a kind of damping fluid measured for human serum glutamyl transferase.
Background technology
Glutamyl transferase (GGT; also known as gamma glutamyl transpeptidase or γ-GGT) be present in human body different tissues; GGT in serum is mainly from liver and gall; when liver diseases or liver damage; GGT all can be caused to raise, and therefore GGT is usually used in the diagnosis of the diseases such as obstructive jaundice, cholangitis and cholecystitis clinically.In prior art, it is one of routine clinical zymetology test item carried out that serum GGT gross activity measures.
Current mensuration GGT generally adopts continuous monitoring method, and GGT acts on colourless L-gamma-glutamyl-3-carboxyl-4-nitroaniline, produces yellow paranitroanilinum, monitors continuously, also can use set time colourimetry at wavelength 405 nm.Concrete reaction principle is as follows:
。
Glutamyl in GGT catalytic substrate in sample is transferred to glycylglycine, and generate gamma-glutamyl glycylglycine and 5-amino-2-nitrobenzoate, the generation of the latter causes the rising in wavelength 405 nm place absorbance, and climbing speed is directly proportional to GGT vigor.But normally carrying out of above-mentioned reaction will be ensured, the buffer solution environment that is stable must be had.Damping fluid is the restraining factors determining glutamyl transferase generation enzymatic reaction, is the important component part of this enzymatic determination kit.The preferred buffer measuring serum GGT in patent US3769173 is the tris-HCI buffer of pH 8.2 (Tris-HCl damping fluid); Patent US5474906 A measures GGT and have employed GOOD ' S series of buffer, and working concentration is not from 20 mM ~ 100 mM etc.In addition the conventional damping fluid meeting the required alkali condition of above reaction also has 2-amino-2-methyl-1-propanol (AMP)-hydrochloride buffer, borax-borate buffer, phosphate buffer etc.
Summary of the invention
The technical matters that the present invention mainly solves is to provide a kind of damping fluid measured for human serum glutamyl transferase, makes the kit of compatibility with it in the accuracy and stability of determination data, all be better than the damping fluid of existing report.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is: provide a kind of damping fluid measured for human serum glutamyl transferase, described damping fluid is used in the kit of human serum glutamyl transferase mensuration, described kit comprises the first reagent and the second reagent, the composition of described first reagent consists of: Tris 10 ~ 25 g/L, glycylglycine 15 g/L, trehalose 0.5 g/L, citric acid 1.5 g/L, the composition of described second reagent consists of: Tris 10 ~ 25 g/L, L-gamma-glutamyl-3-carboxyl-4-nitroaniline (GPNA) 4g/L, citric acid 5 g/L, Sodium azide 0.3g/L.
In a preferred embodiment of the present invention, the composition of described first reagent consists of: Tris 24 g/L, glycylglycine 15 g/L, trehalose 0.5 g/L, citric acid 1.5 g/L.
In a preferred embodiment of the present invention, the composition of described second reagent consists of: Tris 24 g/L, L-gamma-glutamyl-paranitroanilinum (GPNA) 4g/L, citric acid 5 g/L, Sodium azide 0.3g/L.
The invention has the beneficial effects as follows: the damping fluid for human serum glutamyl transferase mensuration of the present invention adopts Tris-citric acid as buffer system; with the kit of this damping fluid compatibility not only performance can reach the level of the conventional damping fluid reported at present, and all show more excellent in the accuracy and stability of determination data.Tris-citric acid can be used as the important supplement that human serum glutamyl transferase measures damping fluid kind needed for kit, provides a kind of selection newly, be worth of widely use for people's serum gamma-glutamyl based transferase measures kit buffer system kind.
Embodiment
Be clearly and completely described to the technical scheme in the embodiment of the present invention below, obviously, described embodiment is only a part of embodiment of the present invention, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making other embodiments all obtained under creative work prerequisite, belong to the scope of protection of the invention.
The instrument that below the present invention, embodiment relates to has: UV Blue Star ultraviolet spectrophotometer 1 (Beijing Labtech Instruments Co., Ltd, also or other brand), constant temperature digital display water-bath 1 (Shanghai Mei Xiang Instrument Ltd., also or other brand).
Embodiment one:
There is provided a kind of kit measured for human serum glutamyl transferase, described kit is made up of, using Tris-citric acid as buffer system reagent 1 and reagent 2:
Reagent 1:
Reagent 2:
。
Embodiment two:
There is provided a kind of kit measured for human serum glutamyl transferase, described kit is made up of, using Tris-boric acid as buffer system reagent 1 and reagent 2:
Reagent 1:
Reagent 2:
。
Embodiment three:
There is provided a kind of kit measured for human serum glutamyl transferase, described kit is made up of, using phosphate as buffer system reagent 1 and reagent 2:
Reagent 1:
Reagent 2:
。
Embodiment four:
There is provided a kind of kit measured for human serum glutamyl transferase, described kit is made up of, using borax-boric acid as buffer system reagent 1 and reagent 2:
Reagent 1:
Reagent 2:
。
Embodiment five:
There is provided a kind of kit measured for human serum glutamyl transferase, described kit is made up of, using HEPES-NaOH as buffer system reagent 1 and reagent 2:
Reagent 1:
Reagent 2:
。
Embodiment six:
There is provided a kind of kit measured for human serum glutamyl transferase, described kit is made up of, using AMP-hydrochloric acid as buffer system reagent 1 and reagent 2:
Reagent 1:
Reagent 2:
。
Embodiment seven:
There is provided a kind of kit measured for human serum glutamyl transferase, described kit is made up of, using Tris-HCl as buffer system reagent 1 and reagent 2:
Reagent 1:
Reagent 2:
。
Using above embodiment one to seven reagent preparation as supplying examination kit, using Landau Laboratories, Inc of Britain produce quality controlled serum 2(Quality Control 2), quality controlled serum 3(Quality Control 3) as sample to be tested, ultraviolet-visible pectrophotometer is adopted to detect, if a skip test, two test samples, wherein skip test replaces sample with distilled water, and detailed mensuration process is as shown in the table.The ratio of sample and reagent 1 and reagent 2 is 100 μ L:1000 μ L:250 μ L; the predominant wavelength of setting is 405nm, commplementary wave length is 505nm; 1min reading is postponed after sample and reagent mix; reading duration is 1.5min; draw △ A/min(absorbance change per minute); according to formula GGT(U/L)=△ A/min × F calculates glutamyl transferase in sample (GGT) content, wherein calculated factor F=1900.The target value of quality controlled serum 2 glutamyl transferase is 55U/L, and the target value of quality controlled serum 3 glutamyl transferase is 186U/L, and permissible variation scope is ± 10%.
According to above method of testing, test in every 5 days once, does 3 repetitions at every turn, and monitoring one month, tests 6 times altogether, obtain the test data of each embodiment, refer to following table continuously.Calculate the mean value of 6 measurement results, relative deviation, standard deviation (SD).Relative deviation this can reflect the accuracy of measurement result, standard deviation this can reflect the stability of measurement result.
Following table is the test data of Quality Control 2:
Following table is the test data of Quality Control 3:
From above two table data, embodiment two, embodiment three, embodiment four have exceeded permissible variation scope all for the measured value relative deviation of serum Quality Control 2 and 3, illustrate using Tris-borate buffer system, phosphate buffer, borax-boric acid system and do not reach basic demand as the kit accuracy of buffer system.Relative deviation has embodiment one, embodiment five, embodiment six and embodiment seven within permissible variation scope ± 10%, illustrate that four kinds of buffer systems can make its compatibility kit meet accuracy requirement, wherein embodiment one i.e. Tris-citric acid is as the kit of buffer system, its relative deviation is respectively 5.25% and 3.07%, be in all embodiments closest to target value, illustrate that the kit accuracy using Tris-citric acid as buffer system is better than the kit of other three kinds of buffer system compatibilities.In addition, from standard deviation (SD) data, the numerical value of embodiment one is minimum, be respectively 3.59 and 3.25, illustrate with the kit measurement repeatability of Tris-citric acid buffer system compatibility best, this reagent is more stable, within one month after reagent, can also keep good test performance.Therefore from accuracy and stability angle, Tris-citric acid buffer system can be preferably the buffer system that human serum glutamyl transferase measures kit.
Based on above data and analysis result known; measure kit with the human serum glutamyl transferase of Tris-citric acid buffer system compatibility and all show excellence in accuracy and stability, Tris-citrate buffer solution can measure the important supplement of damping fluid kind needed for kit as human serum glutamyl transferase.Therefore, the buffer system kind that the present invention behaves in serum gamma-glutamyl based transferase mensuration kit composition provides a kind of selection newly, is worth of widely use.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize description of the present invention to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other relevant technical field, be all in like manner included in scope of patent protection of the present invention.
Claims (3)
1. the damping fluid measured for human serum glutamyl transferase, it is characterized in that, described damping fluid is used in the kit of human serum glutamyl transferase mensuration, described kit comprises the first reagent and the second reagent, the composition of described first reagent consists of: Tris 10 ~ 25 g/L, glycylglycine 15 g/L, trehalose 0.5 g/L, citric acid 1.5 g/L, the composition of described second reagent consists of: Tris 10 ~ 25 g/L, L-gamma-glutamyl-3-carboxyl-4-nitroaniline (GPNA) 4g/L, citric acid 5 g/L, Sodium azide 0.3g/L.
2. the damping fluid measured for human serum glutamyl transferase according to claim 1, it is characterized in that, the composition of described first reagent consists of: Tris 24 g/L, glycylglycine 15 g/L, trehalose 0.5 g/L, citric acid 1.5 g/L.
3. the damping fluid measured for human serum glutamyl transferase according to claim 1; it is characterized in that, the composition of described second reagent consists of: Tris 24 g/L, L-gamma-glutamyl-paranitroanilinum (GPNA) 4g/L, citric acid 5 g/L, Sodium azide 0.3g/L.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113278676A (en) * | 2021-04-23 | 2021-08-20 | 深圳市锦瑞生物科技有限公司 | Serum sodium detection reagent and gamma-glutamyl transferase detection reagent |
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| CN1731146A (en) * | 2005-08-11 | 2006-02-08 | 王贤理 | A method for detecting alpha-L-fucosidase vitality and agent therefor |
| CN1995378A (en) * | 2006-12-19 | 2007-07-11 | 北京华大吉比爱生物技术有限公司 | Glutamate-pyruvate transaminase determination method and glutamate-pyruvate transaminase determination reagent kit |
| CN102384893A (en) * | 2011-08-16 | 2012-03-21 | 南京欣迪生物药业工程有限责任公司 | Seminal plasma gamma-L-glutamyl transpeptidase activity quantitative detecting reagent kit and application thereof |
| CN103290097A (en) * | 2013-05-24 | 2013-09-11 | 宁波美康生物科技股份有限公司 | Gamma-glutamoyl transferase detection reagent |
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- 2014-08-14 CN CN201410398200.7A patent/CN104215631A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1731146A (en) * | 2005-08-11 | 2006-02-08 | 王贤理 | A method for detecting alpha-L-fucosidase vitality and agent therefor |
| CN1995378A (en) * | 2006-12-19 | 2007-07-11 | 北京华大吉比爱生物技术有限公司 | Glutamate-pyruvate transaminase determination method and glutamate-pyruvate transaminase determination reagent kit |
| CN102384893A (en) * | 2011-08-16 | 2012-03-21 | 南京欣迪生物药业工程有限责任公司 | Seminal plasma gamma-L-glutamyl transpeptidase activity quantitative detecting reagent kit and application thereof |
| CN103290097A (en) * | 2013-05-24 | 2013-09-11 | 宁波美康生物科技股份有限公司 | Gamma-glutamoyl transferase detection reagent |
Non-Patent Citations (1)
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| 虞啸炫等: "γ-谷氨酰基转移酶参考方法的建立与性能评估", 《检验医学》, vol. 25, no. 9, 30 September 2010 (2010-09-30), pages 693 - 696 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113278676A (en) * | 2021-04-23 | 2021-08-20 | 深圳市锦瑞生物科技有限公司 | Serum sodium detection reagent and gamma-glutamyl transferase detection reagent |
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Application publication date: 20141217 |