CN104198472A - Stable kit for detecting glycation albumin - Google Patents
Stable kit for detecting glycation albumin Download PDFInfo
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- CN104198472A CN104198472A CN201410401260.XA CN201410401260A CN104198472A CN 104198472 A CN104198472 A CN 104198472A CN 201410401260 A CN201410401260 A CN 201410401260A CN 104198472 A CN104198472 A CN 104198472A
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Abstract
The invention provides a stable kit for detecting glycation albumin. The stable kit consists of a reagent R1, a reagent R2 and a reagent R3, wherein the reagent R1 comprises the following components: a buffering solution, glycation amino acid oxidase, albumin protease, peroxidase, catalase, 4-aminoantipyrene, a stabilizing agent and a preservative; the reagent R2 comprises the following components: a buffering solution, albumin protease, peroxidase, chromogen, a stabilizing agent and a preservative; the reagent R3 comprises the following components: a buffering solution, bromocresol green, a surface active agent and a preservative. The stable kit provided by the invention has the advantages that the influence on the detection result caused by endogenous glycation amino acid is removed by adding glycation amino acid oxidase, so that the result is accurate; the stable kit adopts an enzyme method to detect the content of the glycation albumin, and is a simple, fast, sensitive and accurate detection method.
Description
Technical field
The present invention relates to medical immunology in-vitro diagnosis field, particularly, relate to a kind of stable glycosylated albumin detection kit.
Background technology
There is the glycosylation of non-enzymatic in the N-end of glucose in serum and haemocyanin, generates glycated serum protein, and glycosylated albumin accounts for the more than 90% of glycated serum protein.In addition, because albumin stable content in serum exists, globulin can be because the disease factors such as infection cause fluctuation; Therefore Accurate Determining glycosylated albumin to accurate response diabetes glycemic control situation extremely important.
Glycosylated albumin is a reflection index of 2-3 week average blood sugar level in the past.Want shorter than the reflection cycle of blood sugar test " goldstandard " glycosylated hemoglobin.Therefore, glycosylated albumin has advantage than glycosylated hemoglobin aspect the confirmation of result for the treatment of and the adjustment of clinical application amount.In addition, in the situation that many metabolism of hemoglobin are abnormal, the result of glycosylated hemoglobin is affected and can not truly reflects patient's blood sugar level, and the result of glycosylated albumin is unaffected, as blood sugar test of diabetic nephropathy dialysis patient, anaemia patient, pregnancy women etc.
Diagnostic method: as far back as the eighties in last century, Japanese scholars been has just has been researched and developed high pressure liquid phase ion exchange process (HPLC method) and carried out glycosylated albumin mensuration.HPLC method is measured glycosylated albumin can accurately detect patient's aggregate level of glycemic control in a short time, but at that time because its cost is very high, processes sample size little, is not suitable for routine clinical development.
Afterwards, along with the development of Enzyme-multiplied immune technique, the method for immunization mensuration glycosylated albumin starts to be used indirectly.Identify nonglycosylated specific glycosylation region by adopting monoclonal antibody specific high degree of specificity, measure the non-saccharification serum albumin levels of this ad-hoc location, indirectly obtain the content of glycosylated albumin, though the method cost is lower, but measurement result is not accurate enough, in operating process, personal error is large.
Summary of the invention
For defect of the prior art, the object of this invention is to provide a kind of stable glycosylated albumin detection kit.
The present invention is achieved by the following technical solutions:
The present invention relates to a kind of stable glycosylated albumin detection kit, comprise reagent R1, reagent R2 and reagent R3;
Each component and the concentration of described reagent R1 comprise:
The each component of described reagent R2 and concentration comprise:
The each component of described reagent R3 and concentration comprise:
Preferably, the damping fluid in described reagent R1 comprises one or more in MES damping fluid, Good ' s damping fluid, Tris damping fluid, HEPES damping fluid, phthalic acid-hydrochloride buffer.
Preferably, the stabilizing agent in described reagent R1 comprises the one in bovine serum albumin(BSA), trehalose, sucrose, sequestrant.
Preferably, the antiseptic in described reagent R1 is Sodium azide or proclin300.
Preferably, the damping fluid in described reagent R2 comprises one or more in MES damping fluid, Good ' s damping fluid, Tris damping fluid, HEPES damping fluid, phthalic acid-hydrochloride buffer.
Preferably, the chromogen in described reagent R2 comprises one or more in MEHA, TBHB, TOOS.
Preferably, the stabilizing agent in described reagent R2 comprises the one in bovine serum albumin(BSA), trehalose, sucrose or sequestrant.
Preferably, the antiseptic in described reagent R2 is Sodium azide or proclin300.
Preferably, the damping fluid in described reagent R3 comprises one or more in citrate buffer solution, succinic acid damping fluid, Potassium Hydrogen Phthalate-hydrochloride buffer, phthalic acid-hydrochloride buffer.
Preferably, the surfactant in described reagent R3 comprises one or more in polysorbas20, Brij35, Qu Latong-100; Antiseptic in described reagent R3 is Sodium azide or proclin300.
The present invention detects principle:
(1) mensuration of glycosylated albumin:
In sample, first inject glycated amino acid oxidase (KAOD) and have an effect and endogenous glycated amino acid is become to glucosone, amino acid and hydrogen peroxide and remove.In treating fluid, inject the proteinase of dialogue protein-specific, have an effect and generate glycated amino acid from glycosylated albumin.Secondly, inject glycated amino acid oxidase and generate glucosone, amino acid and hydrogen peroxide from glycated amino acid.The hydrogen peroxide generating, under the coexisting of chromogen and 4-AA, under the effect of peroxidase (POD), is varied to bluish violet pigment quantitatively.The glycated amino acid quantitatively generating from glycosylated albumin by measuring the absorbance of bluish violet pigment.
(2) albuminous mensuration:
This law adopts bromcresol green dyestuff colourimetry, and under the condition of pH4.2, the albumin in sample and bromcresol green coupling generate blue-green complex compound, thereby causes the rising of 630nm place absorbance.
Compared with prior art, the present invention has following beneficial effect:
1, the present invention adopts enzyme process to detect the content of glycosylated albumin, is a kind of simple, quick, sensitive, detection method accurately.
2, the present invention is by adding glycated amino acid oxidase to remove the impact of endogenous glycated amino acid on testing result.Make result more accurate.
Detect by the present invention, the result obtaining is glycosylated albumin and 2 groups of numerical value of albumin.The testing result that finally meets clinical requirement is glycosylated albumin result/albumin result gained number percent.
Brief description of the drawings
By reading the detailed description of non-limiting example being done with reference to the following drawings, it is more obvious that other features, objects and advantages of the present invention will become:
Fig. 1 is according to standard items, the calibration curve figure of calibration GA reagent, and wherein X-axis represents the content of GA, Y-axis represents absorbance.
Fig. 2 is according to standard items, the calibration curve figure of calibration ALB reagent, and wherein X-axis represents the content of ALB, Y-axis represents absorbance.
Fig. 3 adopts respectively the GA reagent of reagent of the present invention and Japanese Asahi Kasei Corporation to carry out correlation analysis figure to measured value, what wherein X-axis represented is patients serum's result that reagent of the present invention is measured, what Y-axis represented is patients serum's result that Japanese Asahi Kasei Corporation reagent is measured, coefficient R
2=0.9981, regression equation is y=1.0349x+0.1527.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.Following examples will contribute to those skilled in the art further to understand the present invention, but not limit in any form the present invention.It should be pointed out that to those skilled in the art, without departing from the inventive concept of the premise, can also make some distortion and improvement.These all belong to protection scope of the present invention.
embodiment 1～3
The present embodiment 1～3 relates to stable glycosylated albumin detection kit, is made up of reagent R1, reagent R2 and reagent R3, and each component and the concentration of described reagent R1, R2, R3 are shown in Table 1:
Table 1
The glycosylated albumin detection kit that embodiment of the present invention 1-3 describes, is applicable to various types of full automatic biochemical apparatus, and taking Hitachi's 7170 full automatic biochemical apparatus as example, it operates as table 2, table 3.
Glycosylated albumin analytical approach: Two point end assay, i.e. reagent R1; The consumption of R2 is respectively 240ul and 60ul, sample size 3ul; 240ul reagent R1 adds 3ul sample after 37 DEG C of 5min, to read an A1, adds 60ulR2, after 5min, reads an A2; Detect wavelength predominant wavelength 546nm, commplementary wave length 700nm respectively.
Albumin analytical approach: some end-point methods, the consumption of reagent R3 is 300ul, sample size 3ul; 300ul reagent R1 adds 3ul sample to read absorbance after 37 DEG C of 2min; Detect wavelength predominant wavelength 600nm, commplementary wave length 700nm respectively.
Adopt this reagent and said determination method, the GA that employing Hitachi 7170 Biochemical Analyzers record and the curve (as shown in Figure 1 and Figure 2) of ALB standard items, wherein X-axis represents GA, ALB content (g/L); Y-axis represents absorbance.
Table 2
The mensuration of glycosylated albumin:
Table 3
Albuminous mensuration:
Accuracy test
This experiment purpose is to detect the stability of reagent.
Adopt experimental example 1 reagent, contrast agents, standard items, quality-control product.
Operation steps: use standard items calibration, measure Quality Control 3 times.
Result is resolved: according to detecting data, calculate test average and the deviation with Quality Control value.
The demonstration of table 4 result, the deviation of reagent measurement result of the present invention and Quality Control value is all less than 3%, shows that reagent accuracy of the present invention is very good.
Table 4
The correlation test of reagent
Use the GA reagent of this law invention reagent (specifically filling a prescription with embodiment 1) and contrast agents Asahi Kasei Corporation, 50 parts of human serums (comprising normal and monstrosity) are measured by each autoregressive parameter simultaneously, measured value is carried out to correlation analysis.Measurement result is shown in Fig. 3, X, and Y-axis is measured value (the content % of GA),
Result by Fig. 3 finds out, the relevant of two kinds of reagent is R
2=0.9981, regression equation is y=1.0349x+0.1527.It is good that result shows that this reagent and import reagent are measured patients serum's correlativity, has good specificity and accuracy.
In addition, above experiment is that 7170 full automatic biochemical apparatus that adopt Hitachi, Ltd to manufacture carry out, but reagent of the present invention is not limited to above-mentioned instrument, is also applicable to other full-automatic or semi-automatic biochemical analyzers.
Above specific embodiments of the invention are described.It will be appreciated that, the present invention is not limited to above-mentioned specific implementations, and those skilled in the art can make various distortion or amendment within the scope of the claims, and this does not affect flesh and blood of the present invention.
Claims (10)
1. a stable glycosylated albumin detection kit, is characterized in that, comprises reagent R1, reagent R2 and reagent R3;
Each component and the concentration of described reagent R1 comprise:
The each component of described reagent R2 and concentration comprise:
The each component of described reagent R3 and concentration comprise:
2. stable glycosylated albumin detection kit as claimed in claim 1, it is characterized in that, the damping fluid in described reagent R1 comprises one or more in MES damping fluid, Good ' s damping fluid, Tris damping fluid, HEPES damping fluid, phthalic acid-hydrochloride buffer.
3. stable glycosylated albumin detection kit as claimed in claim 1, is characterized in that, the stabilizing agent in described reagent R1 comprises the one in bovine serum albumin(BSA), trehalose, sucrose, sequestrant.
4. stable glycosylated albumin detection kit as claimed in claim 1, is characterized in that, the antiseptic in described reagent R1 is Sodium azide or proclin300.
5. stable glycosylated albumin detection kit as claimed in claim 1, it is characterized in that, the damping fluid in described reagent R2 comprises one or more in MES damping fluid, Good ' s damping fluid, Tris damping fluid, HEPES damping fluid, phthalic acid-hydrochloride buffer.
6. stable glycosylated albumin detection kit as claimed in claim 1, is characterized in that, the chromogen in described reagent R2 comprises one or more in MEHA, TBHB, TOOS.
7. stable glycosylated albumin detection kit as claimed in claim 1, is characterized in that, the stabilizing agent in described reagent R2 comprises bovine serum albumin(BSA), trehalose, sucrose or sequestrant.
8. stable glycosylated albumin detection kit as claimed in claim 1, is characterized in that, the antiseptic in described reagent R2 is Sodium azide or proclin300.
9. stable glycosylated albumin detection kit as claimed in claim 1, it is characterized in that, the damping fluid in described reagent R3 comprises one or more in citrate buffer solution, succinic acid damping fluid, Potassium Hydrogen Phthalate-hydrochloride buffer, phthalic acid-hydrochloride buffer.
10. stable glycosylated albumin detection kit as claimed in claim 1, is characterized in that, the surfactant in described reagent R3 is one or more in polysorbas20, Brij35, Qu Latong-100; Antiseptic in described reagent R3 is Sodium azide or proclin300.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673878A (en) * | 2015-03-13 | 2015-06-03 | 温州医科大学 | Kit for measuring concentration ratio of glycated albumin and albumin by virtue of single system |
| CN105241830A (en) * | 2015-09-23 | 2016-01-13 | 广州金域医学检验中心有限公司 | Glycolated serum albumin detection reagent and application thereof |
| CN106226425A (en) * | 2016-07-15 | 2016-12-14 | 首都医科大学附属北京朝阳医院 | Serum glycated albumin detection method and special candidate criteria material thereof |
| CN107870170A (en) * | 2017-12-25 | 2018-04-03 | 广州市进德生物科技有限公司 | A kind of kit of luminol chemiluminescence analysis measure glycated albumin |
| CN109991426A (en) * | 2019-04-03 | 2019-07-09 | 深圳市安帝宝科技有限公司 | A kind of glycated albumin detection kit |
| CN114112595A (en) * | 2021-12-03 | 2022-03-01 | 威海威高生物科技有限公司 | Preparation method of fructosamine determination kit liquid calibration product |
| CN114480565A (en) * | 2021-12-24 | 2022-05-13 | 桂林优利特医疗电子有限公司 | Stable high-sensitivity glycated albumin determination kit with strong anti-interference capability |
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| CN101226198A (en) * | 2007-01-16 | 2008-07-23 | 温州市第三人民医院 | Enzymatical detection method of saccharify blood albumen as well as liquid stabilising agent |
| CN201689054U (en) * | 2010-05-27 | 2010-12-29 | 上海执诚生物技术有限公司 | Kit for detecting glycated albumin content in blood |
| CN102565420A (en) * | 2011-12-26 | 2012-07-11 | 宁波美康生物科技股份有限公司 | Human serum glycated albumin array kit |
| CN102914656A (en) * | 2012-07-23 | 2013-02-06 | 四川省新成生物科技有限责任公司 | Detection kit for saccharifying serum albumin by using indirect immunifaction and measuring method |
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2014
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101226198A (en) * | 2007-01-16 | 2008-07-23 | 温州市第三人民医院 | Enzymatical detection method of saccharify blood albumen as well as liquid stabilising agent |
| CN201689054U (en) * | 2010-05-27 | 2010-12-29 | 上海执诚生物技术有限公司 | Kit for detecting glycated albumin content in blood |
| CN102565420A (en) * | 2011-12-26 | 2012-07-11 | 宁波美康生物科技股份有限公司 | Human serum glycated albumin array kit |
| CN102914656A (en) * | 2012-07-23 | 2013-02-06 | 四川省新成生物科技有限责任公司 | Detection kit for saccharifying serum albumin by using indirect immunifaction and measuring method |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673878A (en) * | 2015-03-13 | 2015-06-03 | 温州医科大学 | Kit for measuring concentration ratio of glycated albumin and albumin by virtue of single system |
| CN105241830A (en) * | 2015-09-23 | 2016-01-13 | 广州金域医学检验中心有限公司 | Glycolated serum albumin detection reagent and application thereof |
| CN106226425A (en) * | 2016-07-15 | 2016-12-14 | 首都医科大学附属北京朝阳医院 | Serum glycated albumin detection method and special candidate criteria material thereof |
| CN106226425B (en) * | 2016-07-15 | 2019-04-23 | 首都医科大学附属北京朝阳医院 | Serum glycated albumin detection method and its dedicated candidate criteria substance |
| CN107870170A (en) * | 2017-12-25 | 2018-04-03 | 广州市进德生物科技有限公司 | A kind of kit of luminol chemiluminescence analysis measure glycated albumin |
| CN109991426A (en) * | 2019-04-03 | 2019-07-09 | 深圳市安帝宝科技有限公司 | A kind of glycated albumin detection kit |
| CN114112595A (en) * | 2021-12-03 | 2022-03-01 | 威海威高生物科技有限公司 | Preparation method of fructosamine determination kit liquid calibration product |
| CN114480565A (en) * | 2021-12-24 | 2022-05-13 | 桂林优利特医疗电子有限公司 | Stable high-sensitivity glycated albumin determination kit with strong anti-interference capability |
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