CN106018303A - Kit for testing alpha-L-fucosidase - Google Patents
Kit for testing alpha-L-fucosidase Download PDFInfo
- Publication number
- CN106018303A CN106018303A CN201610544728.XA CN201610544728A CN106018303A CN 106018303 A CN106018303 A CN 106018303A CN 201610544728 A CN201610544728 A CN 201610544728A CN 106018303 A CN106018303 A CN 106018303A
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- China
- Prior art keywords
- fucosidase
- mmol
- alpha
- detection
- test kit
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The invention relates to the technical fields of medical science and biochemistry, in particular to a kit for testing alpha-L-fucosidase. Existing alpha-L-fucosidase detection methods have the defects that detection cost is high, operation is complex and time-consuming, and detection can only be conducted in a qualitative or semiquantitative mode. The kit for testing alpha-L-fucosidase can solve the problems. According to the technical scheme, the reagent of the kit comprises citric acid of 30-90 mmol/L, sodium chloride of 60-100 mmol/L, sodium azide of 0.1-0.6 g/L, EDTA of 12-26 mmol/L, 2-chloro-4-nitrobenzene-alpha-L-fucosidase of 18-72 mmol/L and purified water serving as solvent. The kit can generate a specific hydrolysis product, the reagent and the product are free of toxicity and harm, reaction is quick and sensitive, detection is more accurate, speed is high, and pollution is avoided; detection can be conducted directly on various biochemical analyzers, operation is easy and convenient, no expensive equipment is required, and detection cost is reduced.
Description
Technical field
The present invention relates to medical science and technological field of biochemistry, a kind of examination measuring alpha-L-fucosidase
Agent box.
Background technology
Alpha-L-fucosidase (AFU) is a kind of lysosomal acid hydrolytic enzyme, is widely present in human tissue cell, blood
With in body fluid, the metabolism of involved in sugar albumen, glycolipid and oligosaccharide.Serum in patients with primary hepatic increases, is primary hepatocarcinoma
One of mark.
The serum in patients with primary hepatic AFU level of 81.2% increases, and can improve primary hepatocarcinoma with AFP joint-detection and examine
Disconnected positive rate reaches 93.1%.Dynamically observe judging that therapeutic efficacy for hepatic carcinoma, prognosis, recurrence are significant.Serum AFU is transitivity liver
Cancer, pulmonary carcinoma, breast carcinoma, ovarian cancer, uterus carcinoma also can increase；Also slight increasing is had at liver cirrhosis, chronic hepatitis, digestive tract hemorrhage etc.
High.
Alpha-L-fucosidase clinical meaning:
Reduce: see the fucose savings disease that heritability alpha-L-fucosidase deficiency causes.
Raise: see primary hepatocarcinoma, secondary liver cancer, liver cirrhosis, acute hepatitis etc..
The method of detection alpha-L-fucosidase mainly has chemiluminescence, enzyme linked immunosorbent assay, colorimetry at present.Chemistry
Luminous detection is relatively costly, and the instrument price needed for detection is expensive；Enzyme linked immunosorbent assay operation complicated and time consumption is long, and to behaviour
Certain professional technique requirement is had as personnel, and may only qualitative or sxemiquantitative, it is impossible to satisfied clinic specifically detects demand；Ratio
Color method is the chromogenic reaction utilizing hydrolyzate, but this method has the restriction of detection method, it is impossible to use continuous monitoring method, so that
There is deviation in the calculating of testing result.
Summary of the invention
The present invention is directed to aforesaid operations length complicated, time-consuming, cost is high, detect inaccurate defect, it is provided that and a kind of mensuration α-
The test kit of L-fucose glycosides enzyme.This test kit can reduce testing cost, shorten the detection time, to have good detection accurate
Degree.
The technical scheme that the present invention solves above-mentioned technical problem and provides is: a kind of reagent measuring alpha-L-fucosidase
Box, this test kit comprises single agents, and composition and the constituent content of described single agents are as follows:
2-hydroxy propane-1,2,3-tricarboxylic acids 30～90 mmol/L；
Sodium chloride 60～100 mmol/L；
Sodium azide 0.1～0.6 g/L；
EDTA 12～26 mmol/L；
2-chloro-4-Nitrobenzol-alpha-L-fucosidase 18～72 mmol/L；
Its solvent is purified water.
Preferably, the preparation steps of described single agents is as follows:
(1) by 2-hydroxy propane-1,2,3-tricarboxylic acids are dissolved in purified water, after stirring, are configured to buffer；
(2) sodium chloride, sodium azide, EDTA, 2-chloro-4-Nitrobenzol-alpha-L-fucosidase are dissolved in the buffering that step (1) prepares
In liquid, stir.
It is further preferred that the component of test kit and constituent content are as follows:
2-hydroxy propane-1,2,3-tricarboxylic acids 30～90 mmol/L；
Sodium chloride 80～90 mmol/L；
Sodium azide 0.1～0.5 g/L；
EDTA 12～22 mmol/L；
2-chloro-4-Nitrobenzol-alpha-L-fucosidase 42～72 mmol/L；
Its solvent is purified water.
One of preferred version as the present invention, the single agents of test kit also includes trehalose, the concentration model of trehalose
Enclosing is 5～25 g/L.
AFU in sample to be tested is catalyzed 2-chloro-4-Nitrobenzol-alpha-L-fucosidase (CNPF), generates 2-chloro-4-Nitrobenzol
Phenol, reaction rate is directly proportional to AFU activity in sample, by measuring the absorbance at 405nm wavelength, can calculate the work of AFU
Property.
Activity (the U/L)=CS of AFU in sample ×(U/L)
In formula:
The absorbance changing value of Δ AU/min testing sample average minute clock
The absorbance changing value of Δ AB/min calibration solution average minute clock
The concentration of AFU in CS calibration solution
The method have technical effect that:
(1) test kit that the present invention provides has only to single reagent, produce the specific corresponding product of hydrolysis, reagent and product without
Poison is harmless, reacts quick and sensitive；So detection is more accurate, speed is fast, pollution-free；Can be directly enterprising in all kinds of biochemistry analyzer
Row detection, operational approach is simple and convenient, it is not necessary to capital intensive equipment, saves testing cost.
(2) preferred, as the preparation steps of each component of test kit of the present invention, prepare buffer in advance, can be follow-up
Each component adds the solution pH value environment that fashionable offer one is stable；Further, this preparation steps is simple to operation.
(3) preferred, each component that test kit of the present invention is comprised, it is limited to a concentration range further, this is dense
Degree scope has more stable reaction environment, reacts fast and stable, beneficially test kit and keeps the quick, accurate of testing result
Really, stable.
(4) as further preferred version, the test kit of the present invention is additionally added trehalose, the concentration range of trehalose
It is maintained at 5～25 g/L, so can keep the activity of alpha-L-fucosidase, it is ensured that the accuracy of testing result.
Accompanying drawing explanation
Fig. 1 is the testing result contrast of this test kit and chemoluminescence method.
Detailed description of the invention
Hereinafter the detailed description of the invention of the present invention is described in detail.It should be appreciated that described herein specifically
Embodiment is merely to illustrate and explains the present invention, is not limited to the present invention.
The invention discloses a kind of test kit measuring alpha-L-fucosidase, this test kit is made up of single agents, bag
The composition and the corresponding content that include are as follows:
2-hydroxy propane-1,2,3-tricarboxylic acids 30～90 mmol/L；
Sodium chloride 60～100 mmol/L；
Sodium azide 0.1～0.6 g/L；
EDTA 12～26 mmol/L；
2-chloro-4-Nitrobenzol-alpha-L-fucosidase 18～72 mmol/L；
Its solvent is purified water.
Preferably, bright also disclosing the preparation steps of single agents in mentioned reagent box, concrete preparation steps is as follows:
(1) by 2-hydroxy propane-1,2,3-tricarboxylic acids are dissolved in purified water, after stirring, are configured to buffer；
(2) sodium chloride, sodium azide, EDTA, 2-chloro-4-Nitrobenzol-alpha-L-fucosidase are dissolved in the buffer that step (1) prepares
In, after stirring.
Although all can get relatively accurate detection result according to the concentration range of each component of test kit of the present invention,
But the accuracy of testing result is not concentrated, it is preferred that the content of each component comprised by test kit retrains as follows further:
2-hydroxy propane-1,2,3-tricarboxylic acids 30～90 mmol/L；
Sodium chloride 80～90 mmol/L；
Sodium azide 0.1～0.5 g/L；
EDTA 12～22 mmol/L；
2-chloro-4-Nitrobenzol-alpha-L-fucosidase 42～72 mmol/L；
Its solvent is purified water.
So, the accuracy of test kit is greatly concentrated, and response speed is also the most steadily.
Owing to the activity of alpha-L-fucosidase is relevant with its residing environment, if because environmental factors causes its activity to become
Change, or cause the testing result of test kit inaccurate.Preferably, test kit is additionally added trehalose, the concentration range of trehalose
It is maintained at 5～25 g/L, so can keep the activity of alpha-L-fucosidase, it is ensured that the accuracy of testing result.
Automatic clinical chemistry analyzer parameter is arranged as shown in table 1.
Table 1
Operate according to table 1 operating parameter, referring to the drawings 1, for the testing result of same detection sample, the present invention and chemiluminescence
Method is compared, and both detected values are very close to, from dependency experimental result, this test kit and chemiluminescence detection result phase
Pass property is fine, dependent equation y=0.9896x+0.5795, R2=0.994, this kit results accuracy as can be seen from the results
Reliably, dependency is good, can apply to the detection of the activity of alpha-L-fucosidase.
Claims (4)
1. measuring a test kit for alpha-L-fucosidase, described test kit comprises single agents, it is characterised in that described
The composition of single agents and constituent content as follows:
2-hydroxy propane-1,2,3-tricarboxylic acids 30～90 mmol/L；
Sodium chloride 60～100 mmol/L；
Sodium azide 0.1～0.6 g/L；
EDTA 12～26 mmol/L；
2-chloro-4-Nitrobenzol-alpha-L-fucosidase 18～72 mmol/L；
Its solvent is purified water.
A kind of test kit measuring alpha-L-fucosidase the most as claimed in claim 1, it is characterised in that described single examination
The preparation steps of agent is as follows:
(1) by 2-hydroxy propane-1,2,3-tricarboxylic acids are dissolved in purified water, after stirring, are configured to buffer；
(2) sodium chloride, sodium azide, EDTA, 2-chloro-4-Nitrobenzol-alpha-L-fucosidase are dissolved in the buffering that step (1) prepares
In liquid, stir.
A kind of test kit measuring alpha-L-fucosidase the most as claimed in claim 1, it is characterised in that: described is single
Composition content is as follows:
2-hydroxy propane-1,2,3-tricarboxylic acids 30～90 mmol/L；
Sodium chloride 80～90 mmol/L；
Sodium azide 0.1～0.5 g/L；
EDTA 12～22 mmol/L；
2-chloro-4-Nitrobenzol-alpha-L-fucosidase 42～72 mmol/L；
Its solvent is purified water.
A kind of test kit measuring alpha-L-fucosidase the most as claimed in claim 1, it is characterised in that: described test kit
Single agents also include that trehalose, the concentration range of described trehalose are 5～25 g/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610544728.XA CN106018303A (en) | 2016-07-12 | 2016-07-12 | Kit for testing alpha-L-fucosidase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610544728.XA CN106018303A (en) | 2016-07-12 | 2016-07-12 | Kit for testing alpha-L-fucosidase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106018303A true CN106018303A (en) | 2016-10-12 |
Family
ID=57109448
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610544728.XA Pending CN106018303A (en) | 2016-07-12 | 2016-07-12 | Kit for testing alpha-L-fucosidase |
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| CN (1) | CN106018303A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112986164A (en) * | 2021-05-18 | 2021-06-18 | 上海执诚生物科技有限公司 | Anti-heparin stable alpha-L-fucosidase detection kit and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63279800A (en) * | 1987-05-13 | 1988-11-16 | Oriental Yeast Co Ltd | Method for improving sensitivity and accuracy of analysis |
| CN1912591A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence measuring method of alpha-L-fucosidase in body fluid |
| US20070072243A1 (en) * | 2003-03-31 | 2007-03-29 | Meikle Peter J | Multiplex screening for lysosomal sotrage disorders (lsds) |
| CN101571537A (en) * | 2009-06-05 | 2009-11-04 | 四川省迈克科技有限责任公司 | Enzymic measuring reagent of alpha-L-fucosidase resistant to heparin interference and using method thereof |
| CN104267178A (en) * | 2014-10-15 | 2015-01-07 | 宁波美康生物科技股份有限公司 | Serum AFU detection kit |
-
2016
- 2016-07-12 CN CN201610544728.XA patent/CN106018303A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63279800A (en) * | 1987-05-13 | 1988-11-16 | Oriental Yeast Co Ltd | Method for improving sensitivity and accuracy of analysis |
| US20070072243A1 (en) * | 2003-03-31 | 2007-03-29 | Meikle Peter J | Multiplex screening for lysosomal sotrage disorders (lsds) |
| CN1912591A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence measuring method of alpha-L-fucosidase in body fluid |
| CN101571537A (en) * | 2009-06-05 | 2009-11-04 | 四川省迈克科技有限责任公司 | Enzymic measuring reagent of alpha-L-fucosidase resistant to heparin interference and using method thereof |
| CN104267178A (en) * | 2014-10-15 | 2015-01-07 | 宁波美康生物科技股份有限公司 | Serum AFU detection kit |
Non-Patent Citations (3)
| Title |
|---|
| 刘凤奎 等: "《临床常用检验与诊断速查》", 31 May 2010 * |
| 王祖莲: "单一试剂连续监测法测定α-L-岩藻糖苷酶", 《云南医药》 * |
| 陈康宁 等: "《小化工生产技术指导》", 31 December 1990 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112986164A (en) * | 2021-05-18 | 2021-06-18 | 上海执诚生物科技有限公司 | Anti-heparin stable alpha-L-fucosidase detection kit and application thereof |
| CN112986164B (en) * | 2021-05-18 | 2021-08-24 | 上海执诚生物科技有限公司 | Anti-heparin stable alpha-L-fucosidase detection kit and application thereof |
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Application publication date: 20161012 |