CN106198415A - A kind of test kit measuring glycolated hemoglobin and preparation method thereof - Google Patents
A kind of test kit measuring glycolated hemoglobin and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of test kit measuring glycolated hemoglobin and preparation method thereof, including reagent R1 independent of each other, reagent R2 and reagent R3 tri-liquid component, including composition and corresponding content be: reagent R1: latex glycine buffer, Macrogol 2000, Tween 80, glycerol, bovine serum albumin, sodium azide, reagent R2: glycine buffer, anti-igg antibody, anti-HbA1c antibody, sodium chloride, bovine serum albumin, sodium azide, reagent R3: hemolysin diluent, preparation method is: prepare reagent according to constituent content；Sample to be tested and reagent R3 are mixed, uses hemolysin diluent that sample to be tested is processed；Sample to be tested after processing mixes with reagent R1 and reagent R2 so that it is fully react；Reacted absorbance difference is measured with automatic clinical chemistry analyzer；The concentration of glycolated hemoglobin in sample is calculated according to absorbance changing value.The present invention has the advantages such as accuracy is high, easy to operate.
Description
Technical field
The present invention relates to medical science and technological field of biochemistry, a kind of reagent measuring glycolated hemoglobin
Box and preparation method thereof.
Background technology
Glycolated hemoglobin is the product that in blood of human body, endoerythrocytic hemoglobin is combined with blood glucose, blood glucose and blood red
It is irreversible reaction that the combination of albumen generates glycolated hemoglobin, and is directly proportional to blood sugar concentration, and keeps about 120 days, institute
Can observe the blood sugar concentration before 120 days, the English code name of glycolated hemoglobin is HbA1c, and glycolated hemoglobin is surveyed
Ping and often can reflect patient's glycemic control situation of nearly 8～12 weeks.
Natural (both the non-glycated) hemoglobin is A0(2 α, 2 β chains), subfraction (HbA1a1, HbA1a2, HbA1b and
HbA1c) being formed because of the free amine group of hemoglobin β chain-N-terminal valine and different carbohydrate glycosylations, these are sub-
Component is generically and collectively referred to as HbA1, and in addition to the N-terminal valine of hemoglobin β chain, in haemoglobin molecule, other free amine groups are also joined
With glycosylation (α chain N-terminal valine, Lysine s-amino groups).
The detection method of relatively broad glycolated hemoglobin is used to be mainly high performance liquid chromatography, parent clinically at present
With chromatography, electrophoresis method etc., the operation of these detection methods is complicated, time-consuming, and needs special instrument, relatively costly, on market
There is also other detection method, but generally there is the defect that accuracy of measurement is low.
Summary of the invention
The technical problem to be solved is to overcome low the lacking of prior art operation complexity, accuracy of measurement
Fall into, and a kind of test kit measuring glycolated hemoglobin and preparation method thereof is provided.
The technical scheme that the present invention solves above-mentioned technical problem and provides is: the invention discloses a kind of mensuration HbAle egg
White test kit, including reagent R1 independent of each other, reagent R2 and reagent R3 tri-liquid component, including composition and corresponding content
For:
Reagent R1:
Latex glycine buffer 15 ~ 185 mmol/L
Polyethylene glycol-2000 5 ~ 25 g/L
Tween 80 1.0 ~ 5.0 ml/L
Glycerol 10 ~ 30 mmol/L
Bovine serum albumin 20 ~ 60 g/L
Sodium azide 0.3 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 50 ~ 150 mmol/L
Anti-igg antibody 5 ~ 16g/L
Anti-HbA1c antibody 8 ~ 15 g/L
Sodium chloride 150 ~ 250 mmol/L
Bovine serum albumin 10 ~ 35 g/L
Sodium azide 0.2 ~ 0.9 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 45 ~ 55mmol/L
Its solvent is purified water.
As preferably, the invention discloses a kind of test kit measuring glycolated hemoglobin, including reagent independent of each other
R1, reagent R2 and reagent R3 tri-liquid component, including composition and corresponding content be:
Reagent R1:
Latex glycine buffer 100 mmol/L
Polyethylene glycol-2000 15 g/L
Tween 80 3.0 ml/L
Glycerol 20 mmol/L
Bovine serum albumin 40 g/L
Sodium azide 0.6 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 100 mmol/L
Anti-igg antibody 10g/L
Anti-HbA1c antibody 10 g/L
Sodium chloride 200 mmol/L
Bovine serum albumin 20 g/L
Sodium azide 0.5 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 50mmol/L
Its solvent is purified water.
As preferably, the compound method of described latex glycine buffer is:
(1) weigh glycine to be dissolved in purified water, stir, be configured to 100 mmol/L glycine buffers standby；
(2) selecting particle diameter is the polystyrene microsphere of 40-500nm, disperses polyphenyl with the glycine buffer obtained by step (1)
Ethylene microsphere, prepares the solution that mass concentration is 1%-5% of contained polystyrene microsphere；
(3) solution prepared in above-mentioned steps (2) adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimides hydrochloric acid
Salt, wherein adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimide hydrochloride of 0.5 mg, 20 in every milliliter of solution
React 2 hours under conditions of DEG C-37 DEG C, use centrifuge, under the rotating speed of 15000 rmp/min centrifugal 30 minutes, abandon supernatant,
Again precipitation is dissolved in 100 mmol/L glycine buffers so that the mass concentration of contained polystyrene microsphere is 1%-5%,
Ultrasonic disperse, then re-uses centrifuge, is centrifuged 30 minutes, abandons supernatant, then will precipitate molten under the rotating speed of 15000 rmp/min
In 100 mmol/L glycine buffers so that the mass concentration of contained polystyrene microsphere is 1%-5%, ultrasonic disperse,
Re-use centrifuge, be centrifuged 30 minutes under the rotating speed of 15000 rmp/min, finally remove supernatant, isolated precipitate；
(4) the glycine buffer diluted blood Lactoferrin adsorbent prepared by step (1), prepares contained hemoglobin and inhales
The mass concentration of attached dose is the solution of 1%-5%；
(5) finally step (3) isolated precipitate is dissolved in the solution containing hemoglobin adsorbent obtained by step (4)
In so that till the mass concentration of polystyrene microsphere is 1%-5%, under conditions of 20 DEG C-37 DEG C, close 24 hours i.e.
Latex glycine buffer can be prepared.
As preferably, the invention also discloses preparation method and the user of the test kit of said determination glycolated hemoglobin
Method, comprises the following steps:
A () prepares reagent according to following component content:
Reagent R1:
Latex glycine buffer 15 ~ 185 mmol/L
Polyethylene glycol-2000 5 ~ 25 g/L
Tween 80 1.0 ~ 5.0 ml/L
Glycerol 10 ~ 30 mmol/L
Bovine serum albumin 20 ~ 60 g/L
Sodium azide 0.3 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 50 ~ 150 mmol/L
Anti-igg antibody 5 ~ 16g/L
Anti-HbA1c antibody 8 ~ 15 g/L
Sodium chloride 150 ~ 250 mmol/L
Bovine serum albumin 10 ~ 35 g/L
Sodium azide 0.2 ~ 0.9 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 45 ~ 55mmol/L
Its solvent is purified water；
B sample to be tested and reagent R3 are mixed by (), use hemolysin diluent to process sample to be tested；
Sample to be tested c () will process after mixes with reagent R1 and reagent R2 so that it is fully react；
D () measures reacted absorbance difference with automatic clinical chemistry analyzer；
E () calculates the concentration of glycolated hemoglobin in sample according to absorbance changing value.
As preferably, in step (b), the volume ratio of described sample to be tested and reagent R3 is 1:50.
As preferably, in step (c), described reagent R1 and the volume ratio of reagent R2 are 3:1.
As preferably, in step (c), the volume ratio of the cumulative volume of described sample to be tested and reagent R1 and reagent R2 is 1:
Between 5 to 1:150.
Compared with prior art, the present invention has a following advantageous benefits: the hemoglobin (Hb) in sample and HbAle
Albumen (HbAlc) and polystyrene microsphere generation non-specific adsorption and immobilization, formed after adding HbAlc specific antibody
The complex of latex-HbAlc-anti-human HbAlc antibody, this complex produces certain turbidity with anti-igg antibody generation agglutination,
Its turbidity height HbAlc immobilised with latex surface measures into positive correlation, by measuring turbidity and carrying out with HbAlc calibration curve
Conversion, obtains HbAlc in sample and accounts for the percentage composition of total Hb.
Activity (the %)=C of glycolated hemoglobin in sampleS×(%)
In formula: Δ ATThe sample cell absorbance compared with blank tube absorbance
ΔASThe calibration pipe absorbance compared with blank tube absorbance
CSThe concentration of HbA1c in calibration solution.
The present invention has a following advantageous benefits: with the addition of glycerol in reagent R1 as suspending agent, can fully suspend polyphenyl second
Alkene microsphere, it is to avoid nonspecific coagulation, can be combined rapidly with hemoglobin and glycolated hemoglobin, it is to avoid because polystyrene is micro-
It is insufficient that the coagulation of ball causes test substance to be combined with reagent, improves the sensitivity of reaction.
Detailed description of the invention
Further illustrate below in conjunction with specific embodiment, but the present invention is not limited in these embodiments.
Embodiment 1
The test kit of the present invention includes reagent R1 independent of each other, reagent R2 and reagent R3 tri-liquid component, wherein
Reagent R1:
Latex glycine buffer 100 mmol/L
Polyethylene glycol-2000 15 g/L
Tween 80 3.0 ml/L
Glycerol 20 mmol/L
Bovine serum albumin 40 g/L
Sodium azide 0.6 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 100 mmol/L
Anti-igg antibody 10g/L
Anti-HbA1c antibody 10 g/L
Sodium chloride 200 mmol/L
Bovine serum albumin 20 g/L
Sodium azide 0.5 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 50mmol/L
Its solvent is purified water.
Embodiment 2
The test kit of the present invention includes reagent R1 independent of each other and reagent R2 biliquid component, wherein
Reagent R1:
Latex glycine buffer 15mmol/L
Polyethylene glycol-2000 25 g/L
Tween 80 1.0 ml/L
Glycerol 30 mmol/L
Bovine serum albumin 20g/L
Sodium azide 0.9 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 50 mmol/L
Anti-igg antibody 16g/L
Anti-HbA1c antibody 8 g/L
Sodium chloride 250 mmol/L
Bovine serum albumin 10 g/L
Sodium azide 0.9 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 55mmol/L
Its solvent is purified water.
Embodiment 3
The preparation and application of test kit
1, latex glycine buffer is prepared:
(1) weigh glycine to be dissolved in purified water, stir, be configured to 100 mmol/L glycine buffers standby；
(2) selecting particle diameter is the polystyrene microsphere of 450nm, disperses polyphenyl second with the glycine buffer obtained by step (1)
Alkene microsphere, prepares the solution that mass concentration is 3% of contained polystyrene microsphere；
(3) solution prepared in above-mentioned steps (2) adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimides hydrochloric acid
Salt, wherein adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimide hydrochloride of 0.5 mg, 25 in every milliliter of solution
React 2 hours under conditions of DEG C, use centrifuge, be centrifuged 30 minutes under the rotating speed of 15000 rmp/min, abandon supernatant, then will
Precipitation is dissolved in 100 mmol/L glycine buffers so that the mass concentration of contained polystyrene microsphere is 3%, ultrasonic point
Dissipate, then re-use centrifuge, be centrifuged 30 minutes under the rotating speed of 15000 rmp/min, abandon supernatant, then precipitation is dissolved in 100
In mmol/L glycine buffer so that the mass concentration of contained polystyrene microsphere is 3%, ultrasonic disperse re-uses centrifugal
Machine, is centrifuged 30 minutes under the rotating speed of 15000 rmp/min, finally removes supernatant, isolated precipitate；
(4) the glycine buffer diluted blood Lactoferrin adsorbent prepared by step (1), prepares contained hemoglobin and inhales
The mass concentration of attached dose is the solution of 3%；
(5) finally step (3) isolated precipitate is dissolved in the solution containing hemoglobin adsorbent obtained by step (4)
In so that till the mass concentration of polystyrene microsphere is 3%, closes under conditions of 25 DEG C and can be prepared into for 24 hours
To latex glycine buffer；
2, reagent is prepared according to following component content:
Reagent R1:
Latex glycine buffer 100 mmol/L
Polyethylene glycol-2000 15 g/L
Tween 80 3.0 ml/L
Glycerol 20 mmol/L
Bovine serum albumin 40 g/L
Sodium azide 0.6 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 100 mmol/L
Anti-igg antibody 10g/L
Anti-HbA1c antibody 10 g/L
Sodium chloride 200 mmol/L
Bovine serum albumin 20 g/L
Sodium azide 0.5 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 50mmol/L
Its solvent is purified water；
3, automatic clinical chemistry analyzer parameter is arranged
(a) detection temperature: 37 DEG C；
(b) detection wavelength: dominant wavelength 660nm；
(c) response time: 10min, wherein, incubation time 5min, measure immediately after adding reagent R2 and read absorbance
Read absorbance A 2 after A1,5min, calculate absorbance changes delta A=A2-A1；
(d) the Direction of Reaction: positive reaction；
4, detecting step
A 5.5 μ l samples to be tested and 275 μ l reagent R3 are mixed by (), use hemolysin diluent to carry out sample to be tested
Process；
(b) to step (a) prepared solution in add 210 μ l reagent R1, mix homogeneously；
C solution after mixing is hatched 5min under conditions of 37 DEG C by ()；
D () adds 70 μ l reagent R2, measure immediately and read absorbance A 1, reads absorbance A 2 after 5min, calculates absorbance change
Δ A=A2-A1；
E () is according to activity (the %)=C of glycolated hemoglobin in sampleS × (%) HbAle in sample is calculated
The concentration of albumen.
Embodiment 4
The preparation and application of test kit
1, latex glycine buffer is prepared:
(1) weigh glycine to be dissolved in purified water, stir, be configured to 100 mmol/L glycine buffers standby；
(2) selecting particle diameter is the polystyrene microsphere of 450nm, disperses polyphenyl second with the glycine buffer obtained by step (1)
Alkene microsphere, prepares the solution that mass concentration is 3% of contained polystyrene microsphere；
(3) solution prepared in above-mentioned steps (2) adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimides hydrochloric acid
Salt, wherein adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimide hydrochloride of 0.5 mg, 25 in every milliliter of solution
React 2 hours under conditions of DEG C, use centrifuge, be centrifuged 30 minutes under the rotating speed of 15000 rmp/min, abandon supernatant, then will
Precipitation is dissolved in 100 mmol/L glycine buffers so that the mass concentration of contained polystyrene microsphere is 3%, ultrasonic point
Dissipate, then re-use centrifuge, be centrifuged 30 minutes under the rotating speed of 15000 rmp/min, abandon supernatant, then precipitation is dissolved in 100
In mmol/L glycine buffer so that the mass concentration of contained polystyrene microsphere is 3%, ultrasonic disperse re-uses centrifugal
Machine, is centrifuged 30 minutes under the rotating speed of 15000 rmp/min, finally removes supernatant, isolated precipitate；
(4) the glycine buffer diluted blood Lactoferrin adsorbent prepared by step (1), prepares contained hemoglobin and inhales
The mass concentration of attached dose is the solution of 3%；
(5) finally step (3) isolated precipitate is dissolved in the solution containing hemoglobin adsorbent obtained by step (4)
In so that till the mass concentration of polystyrene microsphere is 3%, closes under conditions of 25 DEG C and can be prepared into for 24 hours
To latex glycine buffer；
2, reagent is prepared according to following component content:
Reagent R1:
Latex glycine buffer 15mmol/L
Polyethylene glycol-2000 25 g/L
Tween 80 1.0 ml/L
Glycerol 30 mmol/L
Bovine serum albumin 20g/L
Sodium azide 0.9 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 50 mmol/L
Anti-igg antibody 16g/L
Anti-HbA1c antibody 8 g/L
Sodium chloride 250 mmol/L
Bovine serum albumin 10 g/L
Sodium azide 0.9 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 55mmol/L
Its solvent is purified water；
3, automatic clinical chemistry analyzer parameter is arranged
(a) detection temperature: 37 DEG C；
(b) detection wavelength: dominant wavelength 660nm；
(c) response time: 10min, wherein, incubation time 5min, measure immediately after adding reagent R2 and read absorbance
Read absorbance A 2 after A1,5min, calculate absorbance changes delta A=A2-A1；
(d) the Direction of Reaction: positive reaction；
4, detecting step
A 5.5 μ l samples to be tested and 275 μ l reagent R3 are mixed by (), use hemolysin diluent to carry out sample to be tested
Process；
(b) to step (a) prepared solution in add 210 μ l reagent R1, mix homogeneously；
C solution after mixing is hatched 5min under conditions of 37 DEG C by ()；
D () adds 70 μ l reagent R2, measure immediately and read absorbance A 1, reads absorbance A 2 after 5min, calculates absorbance change
Δ A=A2-A1；
E () is according to activity (the %)=C of glycolated hemoglobin in sampleS × (%) HbAle in sample is calculated
The concentration of albumen.
The table 1 test kit measuring glycolated hemoglobin obtained by embodiment 1 and the mensuration sugar obtained by embodiment 2
Changing the result that quality-control product 1 is measured by the test kit of hemoglobin respectively, wherein the glycolated hemoglobin in quality-control product 1 is dense
Degree is 5.50 %, and measurement result is shown in Table 1:
Table 1
| 1st time (%) | 2nd time (%) | 3rd time (%) | Average (%) | Deviation (%) | |
| Embodiment 1 | 5.41 | 5.75 | 5.54 | 5.57 | 1.21 |
| Embodiment 2 | 5.27 | 5.79 | 5.78 | 5.61 | 2.06 |
As shown in Table 1, the test kit measuring glycolated hemoglobin obtained by the present invention is to the measurement result deviation of quality-control product 1 relatively
Little, therefore accuracy of measurement is high, and embodiment 1 is optimum selection.
The table 2 test kit measuring glycolated hemoglobin obtained by embodiment 1 and the mensuration saccharifying blood obtained by embodiment 2
The result that quality-control product 2 is measured by the test kit of Lactoferrin respectively, wherein the concentration of the glycolated hemoglobin in quality-control product 2 is
14.00 %, measurement result is shown in Table 2:
Table 2
| 1st time (%) | 2nd time (%) | 3rd time (%) | Average (%) | Deviation (%) | |
| Embodiment 1 | 14.87 | 14.61 | 13.79 | 17.42 | 3.02 |
| Embodiment 2 | 13.18 | 13.13 | 13.42 | 13.24 | 5.40 |
As shown in Table 2, the test kit measuring glycolated hemoglobin obtained by the present invention is to the measurement result deviation of quality-control product 2 relatively
Little, therefore accuracy of measurement is high, and embodiment 1 is optimum selection.
Same sample to be tested is carried out repeatedly by the table 3 test kit measuring glycolated hemoglobin obtained by embodiment 3
Be repeatedly measured and obtained by embodiment 4 measure glycolated hemoglobin test kit same sample to be tested is carried out the most anti-
Repetition measurement is fixed, and the result of gained carries out the calculating of SD and CV, and result is as follows:
Table 3
The precision of the test kit measuring glycolated hemoglobin obtained by the present invention is relatively good as shown in Table 3, and can by table 3
Knowing, embodiment 3 is optimum selection.
The principle of above-described embodiment only illustrative present invention and effect thereof, not for limiting the present invention.Any ripe
Above-described embodiment all can be modified under the spirit and the scope of the present invention or change by the personage knowing this technology.Cause
This, have usually intellectual such as complete with institute under technological thought without departing from disclosed spirit in art
All equivalences become are modified or change, and must be contained by the claim of the present invention.
Claims (7)
1. the test kit measuring glycolated hemoglobin, it is characterised in that: include reagent R1 independent of each other, reagent R2 and examination
Agent R3 tri-liquid component, including composition and corresponding content be:
Reagent R1:
Latex glycine buffer 15 ~ 185 mmol/L
Polyethylene glycol-2000 5 ~ 25 g/L
Tween 80 1.0 ~ 5.0 ml/L
Glycerol 10 ~ 30 mmol/L
Bovine serum albumin 20 ~ 60 g/L
Sodium azide 0.3 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 50 ~ 150 mmol/L
Anti-igg antibody 5 ~ 16g/L
Anti-HbA1c antibody 8 ~ 15 g/L
Sodium chloride 150 ~ 250 mmol/L
Bovine serum albumin 10 ~ 35 g/L
Sodium azide 0.2 ~ 0.9 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 45 ~ 55mmol/L
Its solvent is purified water.
A kind of test kit measuring glycolated hemoglobin the most according to claim 1, it is characterised in that: include independently of one another
Reagent R1, reagent R2 and reagent R3 tri-liquid component, including composition and corresponding content be:
Reagent R1:
Latex glycine buffer 100 mmol/L
Polyethylene glycol-2000 15 g/L
Tween 80 3.0 ml/L
Glycerol 20 mmol/L
Bovine serum albumin 40 g/L
Sodium azide 0.6 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 100 mmol/L
Anti-igg antibody 10g/L
Anti-HbA1c antibody 10 g/L
Sodium chloride 200 mmol/L
Bovine serum albumin 20 g/L
Sodium azide 0.5 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 50mmol/L
Its solvent is purified water.
A kind of test kit measuring glycolated hemoglobin the most according to claim 1 and 2, it is characterised in that: described glue
The compound method of breast glycine buffer is:
(1) weigh glycine to be dissolved in purified water, stir, be configured to 100 mmol/L glycine buffers standby；
(2) selecting particle diameter is the polystyrene microsphere of 40-500nm, disperses polyphenyl with the glycine buffer obtained by step (1)
Ethylene microsphere, prepares the solution that mass concentration is 1%-5% of contained polystyrene microsphere；
(3) solution prepared in above-mentioned steps (2) adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimides hydrochloric acid
Salt, wherein adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimide hydrochloride of 0.5 mg, 20 in every milliliter of solution
React 2 hours under conditions of DEG C-37 DEG C, use centrifuge, under the rotating speed of 15000 rmp/min centrifugal 30 minutes, abandon supernatant,
Again precipitation is dissolved in 100 mmol/L glycine buffers so that the mass concentration of contained polystyrene microsphere is 1%-5%,
Ultrasonic disperse, then re-uses centrifuge, is centrifuged 30 minutes, abandons supernatant, then will precipitate molten under the rotating speed of 15000 rmp/min
In 100 mmol/L glycine buffers so that the mass concentration of contained polystyrene microsphere is 1%-5%, ultrasonic disperse,
Re-use centrifuge, be centrifuged 30 minutes under the rotating speed of 15000 rmp/min, finally remove supernatant, isolated precipitate；
(4) the glycine buffer diluted blood Lactoferrin adsorbent prepared by step (1), prepares contained hemoglobin and inhales
The mass concentration of attached dose is the solution of 1%-5%；
(5) finally step (3) isolated precipitate is dissolved in the solution containing hemoglobin adsorbent obtained by step (4)
In so that till the mass concentration of polystyrene microsphere is 1%-5%, under conditions of 20 DEG C-37 DEG C, close 24 hours i.e.
Latex glycine buffer can be prepared.
The preparation method of a kind of test kit measuring glycolated hemoglobin the most according to claim 1 and 2 and using method,
It is characterized in that: comprise the following steps:
A () prepares reagent according to following component content:
Reagent R1:
Latex glycine buffer 15 ~ 185 mmol/L
Polyethylene glycol-2000 5 ~ 25 g/L
Tween 80 1.0 ~ 5.0 ml/L
Glycerol 10 ~ 30 mmol/L
Bovine serum albumin 20 ~ 60 g/L
Sodium azide 0.3 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Glycine buffer 50 ~ 150 mmol/L
Anti-igg antibody 5 ~ 16g/L
Anti-HbA1c antibody 8 ~ 15 g/L
Sodium chloride 150 ~ 250 mmol/L
Bovine serum albumin 10 ~ 35 g/L
Sodium azide 0.2 ~ 0.9 g/L
Its solvent is purified water
Reagent R3:
Hemolysin diluent 45 ~ 55mmol/L
Its solvent is purified water；
B sample to be tested and reagent R3 are mixed by (), use hemolysin diluent to process sample to be tested；
Sample to be tested c () will process after mixes with reagent R1 and reagent R2 so that it is fully react；
D () measures reacted absorbance difference with automatic clinical chemistry analyzer；
E () calculates the concentration of glycolated hemoglobin in sample according to absorbance changing value.
The preparation method of a kind of test kit measuring glycolated hemoglobin the most according to claim 4 and using method, its
Being characterised by: in step (b), the volume ratio of described sample to be tested and reagent R3 is 1:50.
The preparation method of a kind of test kit measuring glycolated hemoglobin the most according to claim 4 and using method, its
Being characterised by: in step (c), described reagent R1 and the volume ratio of reagent R2 are 3:1.
The preparation method of a kind of test kit measuring glycolated hemoglobin the most according to claim 4 and using method, its
Being characterised by: in step (c), the volume ratio of the cumulative volume of described sample to be tested and reagent R1 and reagent R2 is at 1:5 to 1:150
Between.
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