CN106093419A - A kind of mensuration dimeric test kit of D and preparation method thereof - Google Patents
A kind of mensuration dimeric test kit of D and preparation method thereof Download PDFInfo
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- CN106093419A CN106093419A CN201610358122.7A CN201610358122A CN106093419A CN 106093419 A CN106093419 A CN 106093419A CN 201610358122 A CN201610358122 A CN 201610358122A CN 106093419 A CN106093419 A CN 106093419A
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Abstract
The invention discloses a kind of mensuration dimeric test kit of D and preparation method thereof, including reagent R1 independent of each other and reagent R2 biliquid component: reagent R1:Tris buffer, sodium chloride, PEG 8000, EDTA, bovine serum albumin, sodium azide, reagent R2:Tris buffer, bovine serum albumin, sucrose, sodium azide, latex are coated anti-human D homodimeric antibody, and preparation method is: prepare reagent；Sample to be tested is mixed with reagent R1 and reagent R2 so that it is fully react；Reacted absorbance difference is measured with automatic clinical chemistry analyzer；The dimeric concentration of D calculated in sample according to absorbance changing value.The present invention has accuracy advantages of higher.
Description
Technical field
The present invention relates to medical science and technological field of biochemistry, a kind of test kit measuring DDi and
Its preparation method.
Background technology
Fibrinolytic system is the most important anticoagulation system of human body, is made up of 4 kinds of major parts: plasminogen, fibre
Dissolved preferment activator, fibrinolysin, plasmin inhibitor, when fibrin coagula is formed, activate at histiotype plasminogen
In the presence of thing (tpA), Plasminogen activation is converted into fibrinolysin, and plasmin solution preocess starts, plasmin degradation fiber
Albumen coagula forms various solvable fragment, forms fibrin degradation product (FDP), and fibrin degradation product (FDP) is by following material: X-
Oligomer, DDi, intermediate segment, fragment E form.Wherein, X-oligomer and D-aggressiveness are all containing DDi unit.
Human body fibrinolytic system, its normal permeability to holding blood vessel wall, maintain flow regime and the tissue repair of blood
Playing an important role, DDi increases explanation at plasma levels and there is Secondary cases fibrinolytic process, and first generates thrombin, after
Have again fibrinolytic system to activate, and also be reflected in thrombotic local plasmin activity or concentration exceedes blood plasma 2 ‰-anti-fibre
Antiplasmin activity or concentration, thromboembolism treatment refers to carry out activated fiber fibrinolytic system with medicine, generally puts into a kind of fibrinolysin
Former activator such as Enzyme In Urine, streptokinase or Tissue plasminogen activator (tpA), make a large amount of fibrinolysin generate, thus accelerate
The most thrombosed dissolving, FDP or DDi generate, then show to reach thrombolytic effect.
In fibrinolytic protein catabolite, the thrombolysis activity after only DDi crosslinking fragment can reflect thrombosis, therefore,
In theory, the detection by quantitative of DDi can quantitative response medicine thrombolytic effect, can be used for diagnosing, screening the blood being newly formed
Bolt.
Measure fibrinolytic system Main Factors, for Clinics and Practices fibrinolytic system disease and have related disorders with fibrinolytic system,
Such as tumor, pregnancy syndrome, and thromboembolism treatment monitoring, have great significance.
The level of fibrin degradation product (FDP) D raises, and shows internal to there is fibrin degradation process frequently.Cause
This, fiber DDi is deep venous thrombosis (DVT), pulmonary infarction (PE), the key index of disseminated inravascular coagulation (DIC).
At present, the method for detection DDi has enzyme linked immunosorbent assay, colloidal gold method, but enzyme linked immunosorbent assay is grasped
Make complexity, the longest, colloidal gold method can qualitative or sxemiquantitative, but the most inadequate at quantitative aspect, and above two method
All there is the defect that accuracy 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 DDi 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 DDi
Test kit, including reagent R1 independent of each other and reagent R2 biliquid component, including composition and corresponding content be:
Reagent R1:
Tris buffer 15 ~ 185 mmol/L
Sodium chloride 150 ~ 300 mmol/L
PEG-8 000 5 ~ 25 g/L
EDTA 10~40 mmol/L
Bovine serum albumin 20 ~ 70 g/L
Sodium azide 0.6 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 50 ~ 150 mmol/L
Bovine serum albumin 4 ~ 18 g/L
Sucrose 5 ~ 35 g/L
Sodium azide 0.6 ~ 0.9 g/L
Latex is coated anti-human DDi antibody 1 ~ 4 g/L
Its solvent is purified water.
As preferably, the invention discloses a kind of test kit measuring DDi, including reagent R1 independent of each other and
Reagent R2 biliquid component, including composition and corresponding content be:
Reagent R1:
Tris buffer 100 mmol/L
Sodium chloride 225 mmol/L
PEG-8 000 15 g/L
EDTA 25 mmol/L
Bovine serum albumin 45 g/L
Sodium azide 0.7 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 100 mmol/L
Bovine serum albumin 11 g/L
Sucrose 20 g/L
Sodium azide 0.7 g/L
Latex is coated anti-human DDi antibody 3 g/L
Its solvent is purified water.
As preferably, in described reagent R1, the pH value of described Tris buffer is 8 ~ 10.
As preferably, in described reagent R2, the pH value of described Tris buffer is 8 ~ 10.
As preferably, in described reagent R2, described latex is coated the preparation method of anti-human DDi antibody and is: use
The polystyrene microsphere dilution that particle diameter is 80 ~ 500nm is become contained polystyrene microsphere by the MES buffer of 50 mmol/L
The solution that concentration is 5 g/L, then in every milliliter of solution add 1-ethyl-3-(3-DimethylAminopropyl) carbodiimides salt
Hydrochlorate 1.0 mg, reacts 1 hour under conditions of 20 ~ 37 DEG C, uses centrifuge, centrifugal under the rotating speed of 20000 rpm/min
30 minutes, remove supernatant, precipitation is suspended in the MES buffer of 50 mmol/L, ultrasonic disperse on ultrasonic disperse instrument, then makes
With centrifuge, it is centrifuged 30 minutes under the rotating speed of 20000 rpm/min, abandons supernatant, then precipitation is joined 50 mmol/L's
In MES buffer, till the concentration of polystyrene microsphere is 10 g/L, using ultrasonic disperse instrument ultrasonic disperse, limit is stirred
Limit adds isopyknic MES buffer containing 8 mg/mL anti-human DDi antibody, and mix and blend, under conditions of 20 ~ 37 DEG C
React 2 hours, till the ultimate density of polystyrene microsphere is 5 g/L, then add bovine serum albumin so that cattle
Serum albumin ultimate density reaches 15 g/L, closes 8 hours, can prepare latex and be coated anti-human under conditions of 4 DEG C
DDi antibody.
As preferably, the invention also discloses preparation method and the using method of the test kit of said determination DDi,
Comprise the following steps:
A () prepares reagent according to following component content:
Reagent R1:
Tris buffer 15 ~ 185 mmol/L
Sodium chloride 150 ~ 300 mmol/L
PEG-8 000 5 ~ 25 g/L
EDTA 10~40 mmol/L
Bovine serum albumin 20 ~ 70 g/L
Sodium azide 0.6 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 50 ~ 150 mmol/L
Bovine serum albumin 4 ~ 18 g/L
Sucrose 5 ~ 35 g/L
Sodium azide 0.6 ~ 0.9 g/L
Latex is coated anti-human DDi antibody 1 ~ 4 g/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 DDi in sample according to absorbance changing value.
As preferably, in step (b), the volume ratio of described reagent R1 and reagent R2 is 4:1.
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 1:30.
The Cleaning Principle of the present invention is: the present invention utilizes antigen antibody reaction, and in sample, DDi (D-D) can be with reagent
In corresponding specificity anti-D-D antibodies form antigen-antibody complex, produce certain turbidity.This turbidity height is necessarily
In the presence of antibody, the content to antigen is directly proportional.Under certain wavelength, measure turbidity and D-bis-can be carried out by multiple spot calibration curve
The quantitative determination of aggressiveness.
Activity (the mg/L)=C of DDi (D-D) in sampleS × (mg/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 D-D in calibration solution.
Compared with prior art, the present invention has following advantageous benefits: by interpolation sucrose as suspending agent in the present invention,
Make latex be coated anti-human DDi antibody suspend more stable, it is easy to participate in reaction, and with the addition of accelerator Polyethylene Glycol-
8000, even if under relatively low concentration, it is also possible to react rapidly so that detecting sensitiveer, therefore accuracy is higher,
Detection results is more preferable.
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 100 mmol/L
Sodium chloride 225 mmol/L
PEG-8 000 15 g/L
EDTA 25 mmol/L
Bovine serum albumin 45 g/L
Sodium azide 0.7 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 100 mmol/L
Bovine serum albumin 11 g/L
Sucrose 20 g/L
Sodium azide 0.7 g/L
Latex is coated anti-human DDi antibody 3 g/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:
Tris buffer 100 mmol/L
Sodium chloride 150 mmol/L
PEG-8 000 25 g/L
EDTA 20 mmol/L
Bovine serum albumin 25 g/L
Sodium azide 0.6 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 50 mmol/L
Bovine serum albumin 18 g/L
Sucrose 5 g/L
Sodium azide 0.9 g/L
Latex is coated anti-human DDi antibody 4 g/L
Its solvent is purified water.
Embodiment 3
The preparation and application of test kit
1, prepare latex and be coated anti-human DDi antibody:: it is the polyphenyl of 120nm with the MES buffer of 50 mmol/L by particle diameter
The dilution of ethylene microsphere becomes the solution that concentration is 5 g/L of contained polystyrene microsphere, then adds 1-in every milliliter of solution
Ethyl-3-(3-DimethylAminopropyl) carbodiimide hydrochloride 1.0 mg, under conditions of 25 DEG C react 1 hour, use from
Scheming, is centrifuged 30 minutes under the rotating speed of 20000 rpm/min, removes supernatant, and precipitation is suspended in the MES buffering of 50 mmol/L
In liquid, ultrasonic disperse on ultrasonic disperse instrument, re-use centrifuge, be centrifuged 30 minutes under the rotating speed of 20000 rpm/min, abandon
Supernatant, then precipitation is joined in the MES buffer of 50 mmol/L, till the concentration of polystyrene microsphere is 10 g/L,
Use ultrasonic disperse instrument ultrasonic disperse, add isopyknic MES buffering containing 8 mg/mL anti-human DDi antibody while stirring
Liquid, mix and blend, reaction 2 hours under conditions of 25 DEG C, till the ultimate density of polystyrene microsphere is 5 g/L, so
After add bovine serum albumin so that bovine serum albumin ultimate density reaches 15 g/L, closes 8 little under conditions of 4 DEG C
Time, latex can be prepared and be coated anti-human DDi antibody；
2, reagent is prepared according to following component content:
Reagent R1:
Tris buffer 100 mmol/L
Sodium chloride 225 mmol/L
PEG-8 000 15 g/L
EDTA 25 mmol/L
Bovine serum albumin 45 g/L
Sodium azide 0.7 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 100 mmol/L
Bovine serum albumin 11 g/L
Sucrose 20 g/L
Sodium azide 0.7 g/L
Latex is coated anti-human DDi antibody 3 g/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 570nm；
(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 () takes 240 μ l reagent R1 and the mixing of 12 μ l samples to be tested；
B solution after mixing is hatched 5min under conditions of 37 DEG C by ()；
C () adds 60 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 mg/L)=C of DDi in sampleS × (mg/L) the D-dimerization in sample is calculated
The concentration of body.
Embodiment 4
The preparation and application of test kit
1, prepare latex and be coated anti-human DDi antibody:: it is the polyphenyl of 80nm with the MES buffer of 50 mmol/L by particle diameter
The dilution of ethylene microsphere becomes the solution that concentration is 5 g/L of contained polystyrene microsphere, then adds 1-in every milliliter of solution
Ethyl-3-(3-DimethylAminopropyl) carbodiimide hydrochloride 1.0 mg, under conditions of 26 DEG C react 1 hour, use from
Scheming, is centrifuged 30 minutes under the rotating speed of 20000 rpm/min, removes supernatant, and precipitation is suspended in the MES buffering of 50 mmol/L
In liquid, ultrasonic disperse on ultrasonic disperse instrument, re-use centrifuge, be centrifuged 30 minutes under the rotating speed of 20000 rpm/min, abandon
Supernatant, then precipitation is joined in the MES buffer of 50 mmol/L, till the concentration of polystyrene microsphere is 10 g/L,
Use ultrasonic disperse instrument ultrasonic disperse, add isopyknic MES buffering containing 8 mg/mL anti-human DDi antibody while stirring
Liquid, mix and blend, reaction 2 hours under conditions of 26 DEG C, till the ultimate density of polystyrene microsphere is 5 g/L, so
After add bovine serum albumin so that bovine serum albumin ultimate density reaches 15 g/L, closes 8 little under conditions of 4 DEG C
Time, latex can be prepared and be coated anti-human DDi antibody；
2, reagent is prepared according to following component content:
Reagent R1:
Tris buffer 100 mmol/L
Sodium chloride 150 mmol/L
PEG-8 000 25 g/L
EDTA 20 mmol/L
Bovine serum albumin 25 g/L
Sodium azide 0.6 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 50 mmol/L
Bovine serum albumin 18 g/L
Sucrose 5 g/L
Sodium azide 0.9 g/L
Latex is coated anti-human DDi antibody 4 g/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 570nm；
(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 () takes 240 μ l reagent R1 and the mixing of 12 μ l samples to be tested；
B solution after mixing is hatched 5min under conditions of 37 DEG C by ()；
C () adds 60 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 mg/L)=C of DDi in sampleS × (mg/L) the D-dimerization in sample is calculated
The concentration of body.
The table 1 test kit measuring DDi obtained by embodiment 1 and the mensuration D-dimerization obtained by embodiment 2
The result that quality-control product 1 is measured by the test kit of body respectively, wherein the concentration of the DDi in quality-control product 1 is 1.50
Mg/L, measurement result is shown in Table 1:
Table 1
As shown in Table 1, the test kit measuring DDi 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.
Mensuration DDi obtained by the test kit of the table 2 mensuration DDi obtained by embodiment 1 and embodiment 2
The result that quality-control product 2 is measured by test kit respectively, wherein the concentration of the DDi in quality-control product 2 is 3.0 mg/L, surveys
Surely the results are shown in Table 2:
Table 2
| 1st time (mg/L) | 2nd time (mg/L) | 3rd time (mg/L) | Average (mg/L) | Deviation (%) | |
| Embodiment 1 | 3.09 | 2.93 | 2.91 | 2.98 | 0.67 |
| Embodiment 2 | 3.10 | 3.01 | 3.07 | 3.06 | 2.00 |
As shown in Table 2, the test kit measuring DDi 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 DDi test kit same sample to be tested is carried out the most repeatedly
The test kit measuring DDi 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 DDi 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 (8)
1. the test kit measuring DDi, 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:
Tris buffer 15 ~ 185 mmol/L
Sodium chloride 150 ~ 300 mmol/L
PEG-8 000 5 ~ 25 g/L
EDTA 10~40 mmol/L
Bovine serum albumin 20 ~ 70 g/L
Sodium azide 0.6 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 50 ~ 150 mmol/L
Bovine serum albumin 4 ~ 18 g/L
Sucrose 5 ~ 35 g/L
Sodium azide 0.6 ~ 0.9 g/L
Latex is coated anti-human DDi antibody 1 ~ 4 g/L
Its solvent is purified water.
A kind of test kit measuring DDi 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:
Tris buffer 100 mmol/L
Sodium chloride 225 mmol/L
PEG-8 000 15 g/L
EDTA 25 mmol/L
Bovine serum albumin 45 g/L
Sodium azide 0.7 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 100 mmol/L
Bovine serum albumin 11 g/L
Sucrose 20 g/L
Sodium azide 0.7 g/L
Latex is coated anti-human DDi antibody 3 g/L
Its solvent is purified water.
A kind of test kit measuring DDi the most according to claim 1 and 2, it is characterised in that: described reagent R1
In, the pH value of described Tris buffer is 8 ~ 10.
A kind of test kit measuring DDi the most according to claim 1 and 2, it is characterised in that: described reagent R2
In, the pH value of described Tris buffer is 8 ~ 10.
A kind of test kit measuring DDi the most according to claim 1 and 2, it is characterised in that: described reagent R2
In, described latex is coated the preparation method of anti-human DDi antibody and is: with the MES buffer of 50 mmol/L by particle diameter be
The polystyrene microsphere dilution of 80 ~ 500nm becomes the solution that concentration is 5 g/L of contained polystyrene microsphere, then every milli
Rise and solution adds 1-ethyl-3-(3-DimethylAminopropyl) carbodiimide hydrochloride 1.0 mg, the condition of 20 ~ 37 DEG C
Lower reaction 1 hour, uses centrifuge, is centrifuged 30 minutes, removes supernatant, precipitation be suspended under the rotating speed of 20000 rpm/min
In the MES buffer of 50 mmol/L, ultrasonic disperse on ultrasonic disperse instrument, re-use centrifuge, 20000 rpm/min's
It is centrifuged 30 minutes under rotating speed, abandons supernatant, then precipitation is joined in the MES buffer of 50 mmol/L, until polystyrene microsphere
Concentration be 10 g/L till, use ultrasonic disperse instrument ultrasonic disperse, add while stirring and isopyknic contain the 8 anti-human D-of mg/mL
The MES buffer of homodimeric antibody, mix and blend, react 2 hours under conditions of 20 ~ 37 DEG C, until polystyrene microsphere
Till ultimate density is 5 g/L, then add bovine serum albumin so that bovine serum albumin ultimate density reaches 15 g/
L, closes 8 hours under conditions of 4 DEG C, can prepare latex and be coated anti-human DDi antibody.
The preparation method of a kind of test kit measuring DDi 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:
Tris buffer 15 ~ 185 mmol/L
Sodium chloride 150 ~ 300 mmol/L
PEG-8 000 5 ~ 25 g/L
EDTA 10~40 mmol/L
Bovine serum albumin 20 ~ 70 g/L
Sodium azide 0.6 ~ 0.9 g/L
Its solvent is purified water
Reagent R2:
Tris buffer 50 ~ 150 mmol/L
Bovine serum albumin 4 ~ 18 g/L
Sucrose 5 ~ 35 g/L
Sodium azide 0.6 ~ 0.9 g/L
Latex is coated anti-human DDi antibody 1 ~ 4 g/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 DDi in sample according to absorbance changing value.
The preparation method of a kind of test kit measuring DDi the most according to claim 6 and using method, its feature
Being: in step (b), the volume ratio of described reagent R1 and reagent R2 is 4:1.
The preparation method of a kind of test kit measuring DDi the most according to claim 6 and using method, its feature
Being: in step (b), the volume ratio of the cumulative volume of described sample to be tested and reagent R1 and reagent R2 is between 1:5 to 1:30.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110702895A (en) * | 2018-12-28 | 2020-01-17 | 上海长岛生物技术有限公司 | D-dimer detection reagent and method, preparation method of reagent and application of polyethylene glycol |
| CN111999506A (en) * | 2020-08-20 | 2020-11-27 | 安徽伊普诺康生物技术股份有限公司 | D-dimer detection kit and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6378066A (en) * | 1986-09-22 | 1988-04-08 | Yatoron:Kk | Method for measuring fdp |
| JP2000193663A (en) * | 1998-12-25 | 2000-07-14 | Kyokuto Seiyaku Kogyo Kk | Measuring reagent kit for human d-dimer |
| CN101819202A (en) * | 2010-05-06 | 2010-09-01 | 上海太阳生物技术有限公司 | D-Dimer measuring kit (latex immunonephelometry method) |
| JP4589756B2 (en) * | 2005-02-25 | 2010-12-01 | シスメックス株式会社 | D-dimer measurement kit |
| CN102788878A (en) * | 2012-08-16 | 2012-11-21 | 北京恩济和生物科技有限公司 | D-dimer detection reagent kit and preparation method thereof |
| CN104459139A (en) * | 2014-12-05 | 2015-03-25 | 重庆中元生物技术有限公司 | D dimer latex-enhanced immunoturbidimetric assay kit utilizing surface functional group |
-
2016
- 2016-05-26 CN CN201610358122.7A patent/CN106093419A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6378066A (en) * | 1986-09-22 | 1988-04-08 | Yatoron:Kk | Method for measuring fdp |
| JP2000193663A (en) * | 1998-12-25 | 2000-07-14 | Kyokuto Seiyaku Kogyo Kk | Measuring reagent kit for human d-dimer |
| JP4589756B2 (en) * | 2005-02-25 | 2010-12-01 | シスメックス株式会社 | D-dimer measurement kit |
| CN101819202A (en) * | 2010-05-06 | 2010-09-01 | 上海太阳生物技术有限公司 | D-Dimer measuring kit (latex immunonephelometry method) |
| CN102788878A (en) * | 2012-08-16 | 2012-11-21 | 北京恩济和生物科技有限公司 | D-dimer detection reagent kit and preparation method thereof |
| CN104459139A (en) * | 2014-12-05 | 2015-03-25 | 重庆中元生物技术有限公司 | D dimer latex-enhanced immunoturbidimetric assay kit utilizing surface functional group |
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Application publication date: 20161109 |