CN106053838A - Kit for determining transferrin and preparation method thereof - Google Patents
Kit for determining transferrin and preparation method thereof Download PDFInfo
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- CN106053838A CN106053838A CN201610544746.8A CN201610544746A CN106053838A CN 106053838 A CN106053838 A CN 106053838A CN 201610544746 A CN201610544746 A CN 201610544746A CN 106053838 A CN106053838 A CN 106053838A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6806—Determination of free amino acids
- G01N33/6812—Assays for specific amino acids
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Abstract
The invention discloses a kit for determining transferrin and a preparation method thereof. The kit comprises dual-liquid components of a reagent R1 and a reagent R2 which are independent from each other. The reagent R1 is prepared from a buffer solution, a surface active agent, a corrosion remover and a stabilizer; the reagent R2 is prepared from a buffer solution, a surface active agent, inorganic salt ions, a corrosion remover and transferrin antibodies, and a solvent of the reagent R2 is purified water. The preparation method comprises the steps that the reagents are prepared according to the component content; a to-be-tested sample is mixed with the reagent R1 and the reagent R2 to be subjected to a sufficient reaction; a fully automatic biochemical analyzer is used for determining an absorptivity difference obtained after the reaction; according to an absorptivity change value, the concentration of transferrin in the sample is worked out. The kit has the advantages of being convenient to operate, high in accuracy and the like.
Description
Technical field
The present invention relates to medical science and technological field of biochemistry, a kind of test kit measuring transferrins and
Its preparation method.
Background technology
Transferrins has another name called transferrin (TRF), is iron protein main in blood plasma, is responsible for delivery and is inhaled by digestive tube
The ferrum received and the ferrum discharged by erythrocyte degraded, with TRF-Fe3+Composite form enter in bone marrow, for the life of mature erythrocyte
Becoming, transferrin molecules amount about 7.7 ten thousand, for single chain glycoprotein, sugar content about 6%, TRF reversibly combines multivalent ion, including
Ferrum, copper, zinc, cobalt etc., every a part TRF can be in conjunction with two ferric iron atoms, and TRF is mainly synthesized by hepatocyte, and the half-life is 7
My god.The regulation that in blood plasma, the concentration of TRF is supplied by ferrum, when iron deficiency state, blood plasma TRF concentration rises, after ferrum is effectively treated
Return to normal level.
In blood plasma, Transferrin can be used for the diagnosis of anemia and the monitoring to treatment, lean at the low hematochrome of iron-deficient
In blood, the level of TRF increases, but the saturation of its ferrum is the lowest, on the contrary, if anemia is due to the erythrocyte Use barriers to ferrum,
Then in blood plasma, TRF is normal or low, but the saturation of ferrum increases.When iron load excess, TRF level is normal, but saturation can
More than 50%, even reach 90%.
Acute-phase response often reduces, therefore in inflammation, malignant change often along with albumin, prealbumin together
Time decline, also decline when chronic liver disease and malnutrition, therefore can be as nutritional status a index.
The defect that the reagent measuring transferrins in the market generally exists operation inconvenience, accuracy of measurement is low, because of
These needs improve.
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 transferrins and preparation method thereof is provided.
The technical scheme that the present invention solves above-mentioned technical problem and provides is: the invention discloses and a kind of measures transferrins
Test kit, including reagent R1 independent of each other and reagent R2 biliquid component, including composition and corresponding content be:
Reagent R1:
Buffer 30 ~ 100 mmol/L
Accelerator 10 ~ 35 g/L
Surfactant 8.0 ~ 18.0 ml/L
Preservative 0.1 ~ 0.7 g/L
Stabilizer 10 ~ 30 g/L
Its solvent is purified water
Reagent R2:
Buffer 30 ~ 100 mmol/L
Surfactant 8.0 ~ 18.0 ml/L
Inorganic ion 150 ~ 250 mmol/L
Preservative 0.1 ~ 0.7 g/L
Transferrin antibodies 3.0 ~ 14.0 ml/L
Its solvent is purified water.
As preferably, the invention discloses a kind of test kit measuring transferrins, including reagent R1 independent of each other and
Reagent R2 biliquid component, including composition and corresponding content be:
Reagent R1:
Buffer 65 mmol/L
Accelerator 20 g/L
Surfactant 13 ml/L
Preservative 0.4 g/L
Stabilizer 20 g/L
Its solvent is purified water
Reagent R2:
Buffer 65 mmol/L
Surfactant 13ml/L
Inorganic ion 200 mmol/L
Preservative 0.4 g/L
Transferrin antibodies 8 ml/L
Its solvent is purified water.
As preferably, in described reagent R1, described buffer uses TRIS buffer, MOPS to delay
Rushing the combination of one or more in liquid, PBS, MES buffer, glycine buffer, described accelerator uses poly-
The combination of one or more in ethylene glycol-2000, PEG-8 000, polyvinylpyrrolidone, described surfactant
The one in tween 80, polyoxyethylene phenyl ether, described preservative is used to use in phenol, sodium benzoate, sodium azide
Kind or multiple combination, described stabilizer uses the combination of one or more in bovine serum albumin, sucrose, trehalose.
As preferably, in described reagent R2, described buffer uses TRIS buffer, MOPS to delay
Rushing the combination of one or more in liquid, PBS, MES buffer, glycine buffer, described surfactant is adopted
By the one in tween 80, polyoxyethylene phenyl ether, described inorganic ion uses in sodium chloride, potassium chloride, magnesium chloride
The combination of one or more, described preservative uses the combination of one or more in phenol, sodium benzoate, sodium azide.
As preferably, the invention also discloses preparation method and the using method of the test kit of said determination transferrins,
Comprise the following steps:
A () prepares reagent according to following component content:
Reagent R1:
Buffer 30 ~ 100 mmol/L
Accelerator 10 ~ 35 g/L
Surfactant 8.0 ~ 18.0 ml/L
Preservative 0.1 ~ 0.7 g/L
Stabilizer 10 ~ 30 g/L
Its solvent is purified water
Reagent R2:
Buffer 30 ~ 100 mmol/L
Surfactant 8.0 ~ 18.0 ml/L
Inorganic ion 150 ~ 250 mmol/L
Preservative 0.1 ~ 0.7 g/L
Transferrin antibodies 3.0 ~ 14.0 ml/L
Its solvent is purified water；
B sample to be tested is mixed by () with reagent R1 and reagent R2 so that it is fully react；
C () measures reacted absorbance difference with automatic clinical chemistry analyzer；
D () calculates the concentration of transferrins in sample according to absorbance changing value.
As preferably, in step (b), the volume ratio of described reagent R1 and reagent R2 is 3:1.
As preferably, in step (b), the volume ratio of the cumulative volume of described sample to be tested and reagent R1 and reagent R2 is 1:
Between 5 to 1:200.
The Cleaning Principle of the present invention is: the transferrins in sample can occur special with the transferrin antibodies in reagent R2
Property antigen antibody reaction, form antigenantibody complex, form turbidity, its turbidity height contains with transferrins in sample
Measure into positive correlation.By measuring turbidity and converting with transferrins calibration curve, obtain the content of transferrins in sample.
Activity (the g/L)=C of transferrins in sampleS × (g/L)
In formula: Δ ATThe sample cell absorbance compared with blank tube absorbance
ΔASThe calibration pipe absorbance compared with blank tube absorbance
CSThe concentration of transferrins in calibration solution.
Compared with prior art, the present invention has following advantageous benefits: with the addition of suspending agent and accelerator in this reagent, helps
Suspension is conducive to the dispersion of antibody and stablizes, and accelerator then can accelerate the reaction of antigen-antibody, even if fainter change is also
Can detect, improve reagent analysis sensitivity.
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 and reagent R2 biliquid component, wherein
Reagent R1:
TRIS buffer 65 mmol/L
PEG-8 000 20 g/L
Tween 80 13 ml/L
Sodium azide 0.4 g/L
Bovine serum albumin 20 g/L
Its solvent is purified water
Reagent R2:
TRIS buffer 65 mmol/L
Tween 80 13ml/L
Sodium chloride 200 mmol/L
Sodium azide 0.4 g/L
Transferrin antibodies 8 ml/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:
MOPS buffer 30 mmol/L
Polyethylene glycol-2000 10 g/L
Polyoxyethylene phenyl ether 18.0 ml/L
Sodium benzoate 0.1 g/L
Sucrose 30 g/L
Its solvent is purified water
Reagent R2:
MOPS buffer 30 mmol/L
Tween 80 8.0 ml/L
Potassium chloride 150 mmol/L
Phenol 0.7 g/L
Transferrin antibodies 14.0 ml/L
Its solvent is purified water.
Embodiment 3
The preparation and application of test kit
1, reagent is prepared according to following component content:
Reagent R1:
TRIS buffer 65 mmol/L
PEG-8 000 20 g/L
Tween 80 13 ml/L
Sodium azide 0.4 g/L
Bovine serum albumin 20 g/L
Its solvent is purified water
Reagent R2:
TRIS buffer 65 mmol/L
Tween 80 13ml/L
Sodium chloride 200 mmol/L
Sodium azide 0.4 g/L
Transferrin antibodies 8 ml/L
Its solvent is purified water；
2, automatic clinical chemistry analyzer parameter is arranged
(a) detection temperature: 37 DEG C；
(b) detection wavelength: dominant wavelength 340nm, commplementary wave length 700nm；
(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；
3, detecting step
A () takes 225 μ l reagent R1 and the mixing of 3 μ l samples to be tested；
B solution after mixing is hatched 5min under conditions of 37 DEG C by ()；
C () adds 75 μ l reagent R2, measure immediately and read absorbance A 1, reads absorbance A 2 after 5min, calculates absorbance change
Δ A=A2-A1；
D () is according to activity (the g/L)=C of transferrins in sampleS × (g/L) transferrins in sample is calculated
Concentration.
Embodiment 4
The preparation and application of test kit
1, reagent is prepared according to following component content:
Reagent R1:
MOPS buffer 30 mmol/L
Polyethylene glycol-2000 10 g/L
Polyoxyethylene phenyl ether 18.0 ml/L
Sodium benzoate 0.1 g/L
Sucrose 30 g/L
Its solvent is purified water
Reagent R2:
MOPS buffer 30 mmol/L
Tween 80 8.0 ml/L
Potassium chloride 150 mmol/L
Phenol 0.7 g/L
Transferrin antibodies 14.0 ml/L
Its solvent is purified water；
2, automatic clinical chemistry analyzer parameter is arranged
(a) detection temperature: 37 DEG C；
(b) detection wavelength: dominant wavelength 340nm, commplementary wave length 700nm；
(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；
3, detecting step
A () takes 225 μ l reagent R1 and the mixing of 3 μ l samples to be tested；
B solution after mixing is hatched 5min under conditions of 37 DEG C by ()；
C () adds 75 μ l reagent R2, measure immediately and read absorbance A 1, reads absorbance A 2 after 5min, calculates absorbance change
Δ A=A2-A1；
D () is according to activity (the g/L)=C of transferrins in sampleS × (g/L) transferrins in sample is calculated
Concentration.
The table 1 test kit measuring transferrins obtained by embodiment 1 and the mensuration obtained by embodiment 2 turn ferrum egg
The result that quality-control product 1 is measured by white test kit respectively, wherein the concentration of the transferrins in quality-control product 1 is 1.29 g/
L, measurement result is shown in Table 1:
Table 1
| 1st time (g/L) | 2nd time (g/L) | 3rd time (g/L) | Average (g/L) | Deviation (%) | |
| Embodiment 1 | 1.30 | 1.31 | 1.31 | 1.31 | 1.29 |
| Embodiment 2 | 1.32 | 1.31 | 1.35 | 1.33 | 2.84 |
As shown in Table 1, the test kit measuring transferrins obtained by the present invention is less to the measurement result deviation of quality-control product 1,
Therefore accuracy of measurement is high, and embodiment 1 is optimum selection.
The table 2 test kit measuring transferrins obtained by embodiment 1 and the mensuration obtained by embodiment 2 turn ferrum
The result that quality-control product 2 is measured by the test kit of albumen respectively, wherein the concentration of the transferrins in quality-control product 2 is 3.38
G/L, measurement result is shown in Table 2:
Table 2
| 1st time (g/L) | 2nd time (g/L) | 3rd time (g/L) | Average (g/L) | Deviation (%) | |
| Embodiment 1 | 3.29 | 3.31 | 3.46 | 3.36 | 0.69 |
| Embodiment 2 | 3.32 | 3.30 | 3.39 | 3.34 | 1.28 |
As shown in Table 2, the test kit measuring transferrins obtained by the present invention is less to the measurement result deviation of quality-control product 2,
Therefore accuracy of measurement is high, and embodiment 1 is optimum selection.
Table 3 obtained by embodiment 3 measure transferrins test kit same sample to be tested is carried out the most repeatedly
The test kit measuring transferrins obtained by mensuration and embodiment 4 is repeatedly repeatedly measured what same sample to be tested was carried out,
Result to gained carries out the calculating of SD and CV, and result is as follows:
Table 3
The precision of the test kit measuring transferrins obtained by the present invention is relatively good as shown in Table 3, and as shown in Table 3,
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 transferrins, it is characterised in that: include reagent R1 independent of each other and reagent R2 biliquid
Component, including composition and corresponding content be:
Reagent R1:
Buffer 30 ~ 100 mmol/L
Accelerator 10 ~ 35 g/L
Surfactant 8.0 ~ 18.0 ml/L
Preservative 0.1 ~ 0.7 g/L
Stabilizer 10 ~ 30 g/L
Its solvent is purified water
Reagent R2:
Buffer 30 ~ 100 mmol/L
Surfactant 8.0 ~ 18.0 ml/L
Inorganic ion 150 ~ 250 mmol/L
Preservative 0.1 ~ 0.7 g/L
Transferrin antibodies 3.0 ~ 14.0 ml/L
Its solvent is purified water.
A kind of test kit measuring transferrins the most according to claim 1, it is characterised in that: include examination independent of each other
Agent R1 and reagent R2 biliquid component, including composition and corresponding content be:
Reagent R1:
Buffer 65 mmol/L
Accelerator 20 g/L
Surfactant 13 ml/L
Preservative 0.4 g/L
Stabilizer 20 g/L
Its solvent is purified water
Reagent R2:
Buffer 65 mmol/L
Surfactant 13ml/L
Inorganic ion 200 mmol/L
Preservative 0.4 g/L
Transferrin antibodies 8 ml/L
Its solvent is purified water.
A kind of test kit measuring transferrins the most according to claim 1 and 2, it is characterised in that: described reagent R1
In, described buffer uses TRIS buffer, MOPS buffer, PBS, MES buffer, sweet ammonia
The combination of one or more in acid buffer, described accelerator uses Polyethylene glycol-2000, PEG-8 000, poly-second
The combination of one or more in alkene pyrrolidone, described surfactant uses in tween 80, polyoxyethylene phenyl ether
One, described preservative uses the combination of one or more in phenol, sodium benzoate, sodium azide, and described stabilizer is adopted
With the combination of one or more in bovine serum albumin, sucrose, trehalose.
A kind of test kit measuring transferrins the most according to claim 1 and 2, it is characterised in that: described reagent R2
In, described buffer uses TRIS buffer, MOPS buffer, PBS, MES buffer, sweet ammonia
The combination of one or more in acid buffer, described surfactant uses in tween 80, polyoxyethylene phenyl ether
Kind, described inorganic ion uses the combination of one or more in sodium chloride, potassium chloride, magnesium chloride, described preservative
Use the combination of one or more in phenol, sodium benzoate, sodium azide.
The preparation method of a kind of test kit measuring transferrins the most according to claim 1 and 2 and using method, it is special
Levy and be: comprise the following steps:
A () prepares reagent according to following component content:
Reagent R1:
Buffer 30 ~ 100 mmol/L
Accelerator 10 ~ 35 g/L
Surfactant 8.0 ~ 18.0 ml/L
Preservative 0.1 ~ 0.7 g/L
Stabilizer 10 ~ 30 g/L
Its solvent is purified water
Reagent R2:
Buffer 30 ~ 100 mmol/L
Surfactant 8.0 ~ 18.0 ml/L
Inorganic ion 150 ~ 250 mmol/L
Preservative 0.1 ~ 0.7 g/L
Transferrin antibodies 3.0 ~ 14.0 ml/L
Its solvent is purified water；
B sample to be tested is mixed by () with reagent R1 and reagent R2 so that it is fully react；
C () measures reacted absorbance difference with automatic clinical chemistry analyzer；
D () calculates the concentration of transferrins in sample according to absorbance changing value.
The preparation method of a kind of test kit measuring transferrins the most according to claim 5 and using method, its feature
Being: in step (b), the volume ratio of described reagent R1 and reagent R2 is 3:1.
The preparation method of a kind of test kit measuring transferrins the most according to claim 5 and using method, its feature
Be: in step (b), the volume ratio of the cumulative volume of described sample to be tested and reagent R1 and reagent R2 1:5 to 1:200 it
Between.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610544746.8A CN106053838A (en) | 2016-07-12 | 2016-07-12 | Kit for determining transferrin and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610544746.8A CN106053838A (en) | 2016-07-12 | 2016-07-12 | Kit for determining transferrin and preparation method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109307762A (en) * | 2017-07-26 | 2019-02-05 | 艾博生物医药(杭州)有限公司 | A kind of positive control liquid and its configuration method |
| CN111122875A (en) * | 2020-01-02 | 2020-05-08 | 四川纳海川生物科技有限公司 | IV-type collagen reagent detection kit and preparation method thereof |
| CN111289501A (en) * | 2020-02-21 | 2020-06-16 | 安徽大千生物工程有限公司 | Kit for detecting NO based on indirect colorimetric method and preparation and use methods thereof |
| CN114047338A (en) * | 2021-11-10 | 2022-02-15 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109307762A (en) * | 2017-07-26 | 2019-02-05 | 艾博生物医药(杭州)有限公司 | A kind of positive control liquid and its configuration method |
| CN111122875A (en) * | 2020-01-02 | 2020-05-08 | 四川纳海川生物科技有限公司 | IV-type collagen reagent detection kit and preparation method thereof |
| CN111289501A (en) * | 2020-02-21 | 2020-06-16 | 安徽大千生物工程有限公司 | Kit for detecting NO based on indirect colorimetric method and preparation and use methods thereof |
| CN114047338A (en) * | 2021-11-10 | 2022-02-15 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
| CN114047338B (en) * | 2021-11-10 | 2024-03-12 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
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Application publication date: 20161026 |