CN104360072A - D-dimmer latex-reinforced immunonephelometry detection kit cooperatively adopting ion stabilizer and suspension stabilizer - Google Patents
D-dimmer latex-reinforced immunonephelometry detection kit cooperatively adopting ion stabilizer and suspension stabilizer Download PDFInfo
- Publication number
- CN104360072A CN104360072A CN201410736445.6A CN201410736445A CN104360072A CN 104360072 A CN104360072 A CN 104360072A CN 201410736445 A CN201410736445 A CN 201410736445A CN 104360072 A CN104360072 A CN 104360072A
- Authority
- CN
- China
- Prior art keywords
- stabilizer
- latex
- reagent
- dimer
- damping fluid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to an in vitro diagnostic kit and in particular relates to a D-dimmer latex-reinforced immunonephelometry detection kit cooperatively adopting an ion stabilizer and a suspension stabilizer. The D-dimmer latex-reinforced immunonephelometry detection kit cooperatively adopting the ion stabilizer and the suspension stabilizer comprises two components, namely a reagent R1 and a reagent R2, wherein the reagent R1 mainly consists of a buffer solution 1, a stabilizer 1, a preservative 1, a coagulation accelerator and a protective agent 1; the reagent R2 mainly consists of a buffer solution 2, two polystyrene latex microspheres crosslinked with different D-dimmer monoclonal antibodies, a stabilizer 2, a protective agent 2 and a preservative 2, and the stabilizer 2 in the reagent R2 adopts the ion stabilizer and the suspension stabilizer which are used cooperatively. Compared with the prior art, the D-dimmer latex-reinforced immunonephelometry detection kit cooperatively adopting the ion stabilizer and the suspension stabilizer has the advantages that a latex-reinforced immunonephelometry method is adopted, and detection can be carried out under the condition that wavelength is 400-800nm, so that the detection is easier and the D-dimmer latex-reinforced immunonephelometry detection kit cooperatively adopting the ion stabilizer and the suspension stabilizer can be applied in clinical more easily.
Description
Technical field
The present invention relates to external diagnosis reagent case, particularly relate to a kind of ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit.
Background technology
Fibrinolytic system is the most important anticoagulation system of human body, is made up of: plasminogen, plasminogen activator, fibrinolysin, plasmin inhibitor 4 kinds of major parts.When fibrin coagula is formed, under the existence of plasminogen activator, Plasminogen activation is converted into fibrinolysin, plasmin solution preocess starts, plasmin degradation fibrin coagula forms various solvable fragment, form fibrin product (FDP), FDP is made up of X-oligomer, DDi, intermediate segment and fragment E (Fragment E).Wherein, X-oligomer and D-aggressiveness are all containing DDi unit.
Human body fibrinolytic system, it is to the normal permeability keeping vascular wall, and the flow state and the organization restoration that maintain blood play an important role.DDi plasma levels increases explanation and there is Secondary cases fibrinolytic process, and first produces fibrin ferment, after have again fibrinolytic system to activate; And be also reflected in thrombotic local plasmin activity or concentration and exceed the active or concentration of blood plasma 2 ‰-antiplasmin.Thromboembolism treatment refers to and carrys out activated fiber fibrinolytic system with medicine.Be generally and drop into a kind of plasminog en-activating thing as Enzyme In Urine, streptokinase or Tissue plasminogen activator (tpA), a large amount of fibrinolysin is generated, thus accelerate thrombosed dissolving.FDP or DDi generate, and show to reach thrombolytic effect.
In fibrinolytic protein catabolite, only DDi be cross-linked fragment can reflect thrombosis after thrombolysis activity.Therefore, in theory, DDi quantitative detection can the thrombolytic effect and can be used for of quantitative response medicine diagnose, the new thrombus formed of screening.Therefore, fiber DDi is DVT (DVT), pulmonary embolism (PE), the key index of disseminated intravascular coagulation (DIC).Clinically the detection of DDi content be can be used for the diagnosis of following disease: the monitoring etc. of cranial vascular disease, liver diseases, surgery patients, thromboembolism treatment.
The assay method of DDi is a lot, has ELISA, TRIFMA, collaurum, latex enhancing immune turbidimetry etc.Widely used clinically is at present colloidal gold method, and ELISA operation requirements is strict, more time-consuming, and collaurum is as qualitative or semi-quantitative method, for quantitatively, slightly not enough.And latex enhancing immune turbidimetry has fast, quantitatively and the low advantage of cost, the concern of clinician is more and more received.
Summary of the invention
The shortcoming of prior art in view of the above, the object of the present invention is to provide a kind of good stability, detection sensitivity is high, the range of linearity wide D-dimer detection reagent kit, and the preparation method of D-dimer detection reagent kit, for solving the problems of the prior art.
For achieving the above object and other relevant objects, first aspect present invention provide a kind of ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, comprise reagent R1 and reagent R2 two kinds of components, described reagent R1 forms primarily of damping fluid 1, stabilizing agent 1, antiseptic 1, set accelerator and protective agent 1; Described reagent R2 forms primarily of damping fluid 2, the polystyrene latex microspheres of crosslinked D dimer monoclonal antibody, stabilizing agent 2, protective agent 2, antiseptic 2, is wherein connected with covalent cross-linking or physical adsorption way between polystyrene latex microspheres with D dimer antibody.
Preferably, in the present invention, polystyrene latex microspheres is cross-linked two kinds of different D dimer monoclonal antibodies.
D dimer latex enhancing immune in the technical program is than turbid kit, by antibody linked for D dimer on polystyrene latex microspheres surface, when the D dimer in blood and D dimer antibody response, drive polystyrene latex microspheres to assemble and produce certain turbidity, and turbidity and the D dimer content in blood are directly proportional in certain limit, the content of D dimer in blood can be detected with full automatic biochemical apparatus under 400-800nm wavelength.
Preferably, damping fluid 1 in described reagent R1 is selected from one or more in Hepes damping fluid, Tris-HCl damping fluid, MOPS damping fluid, PBS damping fluid, glycine buffer, borate buffer solution, citrate buffer solution, its pH is 6.0 ~ 9.0, and concentration is 25 ~ 500mmol/L; Damping fluid 2 in described reagent R2 is selected from one or more in PBS damping fluid, borate buffer solution, glycine buffer, Hepes damping fluid, GOODS damping fluid, MOPS damping fluid, and its pH is 7.0 ~ 9.0, and concentration is 25 ~ 500mmol/L.
Damping fluid 1 in reagent R1 of the present invention can use the various conventional damping fluid in above-mentioned this area, but in order to make kit, there is better sensitivity and color developing effect, in the R1 of reagent described in the present invention, damping fluid 1 preferably comprises following component: the aqueous solution of stachyose, alum, Fructose Diphosphate, sodium hexametaphosphate and glycocoll, and the total concentration of stachyose, alum, Fructose Diphosphate, sodium hexametaphosphate, glycocoll is 3.5-7.5g/L, the pH value of damping fluid is 7.2-7.6.
Preferably, the concentration of each component in damping fluid 1 is:
The solvent of described damping fluid 1 is water.
Described damping fluid 1 can use the various conventional pH adjusting agent in this area to carry out the adjustment of pH value.
Preferably, the stabilizing agent 1 in described reagent R1 be selected from KCl, NaCl, CaCl one or several, its mass concentration is 0.5% ~ 10%; Stabilizing agent 2 in described reagent R2 adopt ion stabilizer and suspension stabilizer with the use of; Wherein ion stabilizer is NaCl, KCl, Na
2cO3, Na
2sO4 or K
2sO4, suspension stabilizer is PEG8000, sucrose, glycerine or glucose.
Preferably, the antiseptic 1 in described reagent R1 is Sodium azide, thimerosal or ProClin300; Antiseptic 2 in described reagent R2 is Sodium azide, thimerosal or ProClin300.
Preferably, the set accelerator in described reagent R1 is PEG8000 or glucosan.Preferred PEG8000, be because PEG8000 belongs to non-ionic water-soluble polymer, the dissolubility in water is larger, can regulate the viscosity of reagent R1, promotes that antigen and antibody molecule are combined into compound.
Preferably, the surface functional group of the polystyrene latex microballoon in described reagent R2 is amino, carboxyl, hydrazides, aldehyde radical or epoxy radicals, and the particle diameter of polystyrene latex microballoon is at 100 ~ 500nm.Preferred surface functional group is the polystyrene latex microspheres of carboxyl, and this is that the functional group being carboxyl due to polystyrene latex microspheres surface is easy to be increased coupling effect by EDC activation, ensures sensitivity and the accuracy of test result.
Preferably, the D dimer antibody in described reagent R2 be the anti-human D dimer monoclonal antibody of rabbit, mouse-anti people D dimer monoclonal antibody one or several.Preferably two kinds of mouse monoclonal antibodies, not Seeking Truth pairing monoclonal antibody.
Preferably, the protective agent 1 in described reagent R1 is EDTA, and the concentration of described EDTA is 10 ~ 100mmol/L; Protective agent 2 in described reagent R2 is bovine serum albumin(BSA).
Second aspect present invention provide described ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen the preparation method of immunoturbidimetry detection kit, comprise the steps:
(1) preparation of reagent R1:
In damping fluid 1, add stabilizing agent 1, set accelerator, antiseptic 1, protective agent 1, be uniformly mixed, obtain reagent R1;
(2) preparation of reagent R2:
With mark damping fluid, polystyrene latex microspheres being diluted to mass concentration is 1%; add the EDC that mass concentration is 0.01%-0.1% again; at room temperature stirring reaction 60min; reaction terminates rear 15000rpm centrifuge washing to remove unreacted EDC; then D dimer monoclonal antibody is added; stirring reaction 4h at 37 DEG C; add stop buffer 4h cessation reaction again; by centrifugal for the reactant liquor 15000rpm obtained, washing, finally with comprising, the damping fluid 2 of stabilizing agent 2, protective agent 2, antiseptic 2 is resuspended by latex.Another strain monoclonal antibody of same operational label, namely both obtain R2 after ultrasonic after 1:1 mixing.
Third aspect present invention provide described ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen the purposes of immunoturbidimetry detection kit at D dimer detection field.
Compared with prior art, the present invention has following beneficial effect:
1. adopt latex enhancing immune turbidimetry, when the D dimer in blood and D dimer monoclonal antibody reactive, drive polystyrene latex to assemble and produce certain turbidity, and turbidity is directly proportional within the specific limits to the D dimer content in blood, can detect under the wavelength of 400 ~ 800nm, detect more convenient, easily in clinical middle application.
2. the surface functional group of polystyrene latex microspheres is amino, carboxyl, hydrazides or epoxy radicals etc., the functional group on its surface can combine with the amino of antibody surface etc. and form covalent coupling structure, D dimer monoclonal antibody is made firmly to be combined in latex microsphere surface, ensure that the stability of R2, extend the reagent term of validity.
3. employing particle diameter is two kinds of polystyrene latex particles of 100 ~ 500nm, and a kind of have larger particle diameter, adds turbidity when D dimer and D dimer antibody response in blood, thus increases the sensitivity of test reaction; A kind of particle diameter is less, guarantee reagent linear.
4., in preparation process, adopt the single stage method crosslinked latex and antibody that improve, namely ensure that the cross-linking effect of antibody, ensure again the activity of antibody, simplify mode of operation, be conducive to the carrying out produced.
Accompanying drawing explanation
Fig. 1 is the calibration curve of D-dimer detection reagent kit embodiment of the present invention.
Embodiment
Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this instructions can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by embodiments different in addition, and the every details in this instructions also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.
Before further describing the specific embodiment of the invention, should be understood that protection scope of the present invention is not limited to following specific specific embodiments; It is also understood that the term used in the embodiment of the present invention is to describe specific specific embodiments, instead of in order to limit the scope of the invention; In instructions of the present invention and claims, unless explicitly pointed out in addition in literary composition, singulative " ", " one " and " this " comprise plural form.
When embodiment provides numerical range, should be understood that except non-invention is otherwise noted, between two end points of each numerical range and two end points, any one numerical value all can be selected.Unless otherwise defined, all technology used in the present invention are identical with the meaning that those skilled in the art of the present technique understand usually with scientific terminology.Except the concrete grammar used in embodiment, equipment, material, according to those skilled in the art to the grasp of prior art and record of the present invention, any method of prior art that is similar with the method described in the embodiment of the present invention, equipment, material or that be equal to, equipment and material can also be used to realize the present invention.
Unless otherwise indicated, disclosed in the present invention experimental technique, detection method, preparation method all adopt the routine techniques of the molecular biology of the art routine, biological chemistry, chromatin Structure and analysis, analytical chemistry, cell chulture, recombinant DNA technology and association area.These technology are existing in existing document improves explanation, specifically can see the MOLECULAR CLONING:A LABORATORY MANUAL such as Sambrook, Second edition, Cold Spring HarborLaboratory Press, 1989 and Third edition, 2001; Ausubel etc., CURRENT PROTOCOLS INMOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987 and periodic updates; The seriesMETHODS IN ENZYMOLOGY, Academic Press, San Diego; Wolffe, CHROMATINSTRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; METHODSIN ENZYMOLOGY, Vol.304, Chromatin (P.M.Wassarman and A.P.Wolffe, eds.), AcademicPress, San Diego, 1999; With METHODS IN MOLECULAR BIOLOGY, Vol.119, ChromatinProtocols (P.B.Becker, ed.) Humana Press, Totowa, 1999 etc.
Embodiment 1:
A kind of D dimer latex enhancing immune, than turbid detection kit, comprises following preparation process:
(1) preparation of reagent R1:
Taking 1.9g stachyose, 0.5g alum, 1.9g Fructose Diphosphate, 0.3g sodium hexametaphosphate, 1.8g glycocoll, 27g NaCl, 35g PEG8000,0.5g Sodium azide, 10g bovine serum albumin(BSA) and 1.42g EDTA is dissolved in 0.8L distilled water, regulate pH to 7.4, namely fixed molten 1L obtains reagent R1.
(2) preparation of reagent R2:
With pH be 7.4, concentration is that above-mentioned washed polystyrene latex microspheres is diluted to mass concentration is 1% for the PBS damping fluid of 50mmol/L.The EDC of 0.01g is added in the above-mentioned latex dilution of 1L, at room temperature stirring reaction 60min, reaction terminates rear centrifugal and remove unreacted EDC with PBS buffer solution, then D dimer antibody is added, at 37 DEG C after stirring reaction 4h, add stop buffer 4h cessation reaction, by centrifugal for the reactant liquor 15000rpm obtained, washing, finally use 40g/LNaCl, 50g/L PEG8000,0.5g/L Sodium azide, 50g/L bovine serum albumin(BSA), 25mM glycocoll, the damping fluid 2 (aqueous solution) of pH=7.4 is resuspended by latex, namely obtains R2 after ultrasonic.
Embodiment 2:
A kind of D dimer latex enhancing immune, than turbid detection kit, comprises following preparation process:
(1) preparation of reagent R1:
Taking 1.9g stachyose, 0.5g alum, 1.9g Fructose Diphosphate, 0.3g sodium hexametaphosphate, 1.8g glycocoll, 30g NaCl, 50g PEG8000,0.5g Sodium azide, 50g bovine serum albumin(BSA) and 7.1g EDTA is dissolved in 0.8L distilled water, regulate pH to 7.4, namely fixed molten 1L obtains reagent R1.
(2) preparation of reagent R2:
With pH be 6.5, concentration is that polystyrene latex microspheres is diluted to mass concentration is 3% for the MES damping fluid of 100mmol/L.The EDC of 0.01g is added in the above-mentioned latex dilution of 1L, at room temperature stirring reaction 30min, reaction terminates rear centrifugal and remove unreacted EDC with PBS buffer solution, then D dimer antibody is added, after stirred at ambient temperature reaction 8h, add stop buffer 4h cessation reaction, by centrifugal for the reactant liquor 15000rpm obtained, washing, finally use 40g/L NaCl, 50g/LPEG8000,0.5g/L Sodium azide, 50g/L bovine serum albumin(BSA), 25mM glycocoll, the damping fluid 2 (aqueous solution) of pH=7.4 is resuspended by latex, namely obtains R2 after ultrasonic.
Embodiment 3:
A kind of D dimer latex enhancing immune, than turbid detection kit, comprises following preparation process:
(1) preparation of reagent R1:
Taking 1.9g stachyose, 0.5g alum, 1.9g Fructose Diphosphate, 0.3g sodium hexametaphosphate, 1.8g glycocoll, 15g NaCl, 24g PEG8000,0.5g Sodium azide, 10g bovine serum albumin(BSA) and 7.1g EDTA is dissolved in 0.8L distilled water, regulate pH to 7.4, namely fixed molten 1L obtains reagent R1.
(2) preparation of reagent R2:
With pH be 6.0, concentration is that polystyrene latex microspheres is diluted to mass concentration is 2% for the MES damping fluid of 100mmol/L.The EDC of 0.02g is added in the above-mentioned latex dilution of 1L, at room temperature stirring reaction 30min, reaction terminates rear centrifugal and remove unreacted EDC with PBS buffer solution, then D dimer antibody is added, after stirred at ambient temperature reaction 8h, add stop buffer 4h cessation reaction, by centrifugal for the reactant liquor 15000rpm obtained, washing, finally use 40g/L NaCl, 50g/LPEG8000,0.5g/L Sodium azide, 50g/L bovine serum albumin(BSA), 25mM glycocoll, the damping fluid 2 (aqueous solution) of pH=7.4 is resuspended by latex, namely obtains R2 after ultrasonic.
Embodiment 4:
A kind of D dimer latex enhancing immune, than turbid detection kit, comprises following preparation process:
(1) preparation of reagent R1:
Taking 1.9g stachyose, 0.5g alum, 1.9g Fructose Diphosphate, 0.3g sodium hexametaphosphate, 1.8g glycocoll, 9g NaCl, 50g PEG8000,0.5g Sodium azide, 5g bovine serum albumin(BSA) and 1.42g EDTA is dissolved in 0.8L distilled water, regulate pH to 7.6, namely fixed molten 1L obtains reagent R1.
(2) preparation of reagent R2:
With pH be 6.5, concentration is that polystyrene latex microspheres is diluted to mass concentration is 3% for the MES damping fluid of 100mmol/L.The EDC of 0.03g is added in the above-mentioned latex dilution of 1L, at room temperature stirring reaction 30min, reaction terminates rear centrifugal and remove unreacted EDC with MES buffer solution, then D dimer antibody is added, after stirred at ambient temperature reaction 4h, add stop buffer 4h cessation reaction, by centrifugal for the reactant liquor 15000rpm obtained, washing, finally use 40g/L NaCl, 50g/LPEG8000,0.5g/L Sodium azide, 50g/L bovine serum albumin(BSA), 25mM glycocoll, the damping fluid 2 (aqueous solution) of pH=7.4 is resuspended by latex, namely obtains R2 after ultrasonic.
Embodiment 5:
A kind of D dimer latex enhancing immune, than turbid detection kit, comprises following preparation process:
(1) preparation of reagent R1:
Taking 1.9g stachyose, 0.5g alum, 1.9g Fructose Diphosphate, 0.3g sodium hexametaphosphate, 1.8g glycocoll, 27g NaCl, 15g PEG8000,0.5g Sodium azide, 5g bovine serum albumin(BSA) and 1.42g EDTA is dissolved in 0.8L distilled water, regulate pH to 7.0, namely fixed molten 1L obtains reagent R1.
(2) preparation of reagent R2:
With pH be 7.5, concentration is that polystyrene latex microspheres is diluted to mass concentration is 0.5% for the mops damping fluid of 100mmol/L.The EDC of 0.03g is added in the above-mentioned latex dilution of 1L, at room temperature stirring reaction 30min, reaction terminates rear centrifugal and remove unreacted EDC with mops buffer solution, then D dimer antibody is added, after stirred at ambient temperature reaction 8h, add stop buffer 4h cessation reaction, by centrifugal for the reactant liquor 15000rpm obtained, washing, finally use 40g/L NaCl, 50g/L PEG8000,0.5g/L Sodium azide, 50g/L bovine serum albumin(BSA), 25mM glycocoll, the damping fluid 2 (aqueous solution) of pH=7.4 is resuspended by latex, namely obtains R2 after ultrasonic.
Comparative example:
A kind of D dimer latex enhancing immune, than turbid detection kit, comprises following preparation process:
(1) preparation of reagent R1:
Taking 19g Mops, 27g NaCl, 15g PEG8000,0.5g Sodium azide, 5g bovine serum albumin(BSA) and 1.42g EDTA is dissolved in 0.8L distilled water, regulates pH to 7.0, and namely fixed molten 1L obtains reagent R1.
(2) preparation of reagent R2:
With pH be 7.5, concentration is that polystyrene latex microspheres is diluted to mass concentration is 0.5% for the mops damping fluid of 100mmol/L.The EDC of 0.03g is added in the above-mentioned latex dilution of 1L, at room temperature stirring reaction 30min, reaction terminates rear centrifugal and remove unreacted EDC with mops buffer solution, then D dimer antibody is added, after stirred at ambient temperature reaction 8h, add stop buffer 4h cessation reaction, by centrifugal for the reactant liquor 15000rpm obtained, washing, finally use 40g/L NaCl, 50g/L PEG8000,0.5g/L Sodium azide, 50g/L bovine serum albumin(BSA), 25mM glycocoll, the damping fluid 2 (aqueous solution) of pH=7.4 is resuspended by latex, namely obtains R2 after ultrasonic.
Test result:
Reagent Beckman AU480 full automatic biochemical apparatus prepared by above-mentioned 5 embodiments and comparative example is tested, test wavelength is 6000nm, sample this or calibration object 4uL, add the reagent R1 of 120uL again, 37 DEG C of constant temperature 5min, then add 40ul reagent R2, read absorbance A 1 after 20s, hatch 4 points for 37 DEG C and read absorbance A 2 afterwards in 40 seconds, then react absorbance Δ A=A2-A1; First use standard items to carry out multiple spot calibration, and calculate by splines, obtain calibration curve, as shown in Figure 1.Sample is changed by its absorbance, carries out contrast can obtain PCT concentration in sample with typical curve.Carried out sensitivity for analysis, accuracy, accuracy and stability etc. to above-mentioned 5 embodiments and comparative example to verify, the result is as shown in table 1:
Table 1
According to its test result, detection reagent obtained in parameter area of the present invention, all can possess good sensitivity for analysis, accuracy, accuracy and stability, and wherein implementing 4 is optimum embodiment, can improve reagent test effect best.
In sum, the present invention effectively overcomes various shortcoming of the prior art and tool high industrial utilization.
Above-described embodiment is illustrative principle of the present invention and effect thereof only, but not for limiting the present invention.Any person skilled in the art scholar all without prejudice under spirit of the present invention and category, can modify above-described embodiment or changes.Therefore, such as have in art usually know the knowledgeable do not depart from complete under disclosed spirit and technological thought all equivalence modify or change, must be contained by claim of the present invention.
Claims (10)
1. an ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, comprise reagent R1 and reagent R2 two kinds of components, described reagent R1 forms primarily of damping fluid 1, stabilizing agent 1, antiseptic 1, set accelerator and protective agent 1; Described reagent R2 forms primarily of damping fluid 2, the polystyrene latex microspheres of crosslinked D dimer monoclonal antibody, stabilizing agent 2, protective agent 2, antiseptic 2, wherein be connected with covalent cross-linking or physical adsorption way between polystyrene latex microspheres with D dimer antibody, the stabilizing agent 2 in described reagent R2 adopt ion stabilizer and suspension stabilizer with the use of; Wherein ion stabilizer is NaCl, KCl, Na
2cO3, Na
2sO4 or K
2sO4, suspension stabilizer is PEG8000, sucrose, glycerine or glucose.
2. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, it is characterized in that, damping fluid 1 in described reagent R1 is selected from one or more in Hepes damping fluid, Tris-HCl damping fluid, MOPS damping fluid, PBS damping fluid, glycine buffer, borate buffer solution, citrate buffer solution, its pH is 6.0 ~ 9.0, and concentration is 25 ~ 500mmol/L.
3. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, it is characterized in that, damping fluid 2 in described reagent R2 is selected from one or more in PBS damping fluid, borate buffer solution, glycine buffer, Hepes damping fluid, GOODS damping fluid, MOPS damping fluid, its pH is 7.0 ~ 9.0, and concentration is 25 ~ 500mmol/L.
4. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, it is characterized in that, stabilizing agent 1 in described reagent R1 be selected from KCl, NaCl, CaCl one or several, its mass concentration is 0.5% ~ 10%.
5. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, it is characterized in that, the antiseptic 1 in described reagent R1 is Sodium azide, thimerosal or ProClin300; Antiseptic 2 in described reagent R2 is Sodium azide, thimerosal or ProClin300.
6. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, it is characterized in that, the set accelerator in described reagent R1 is PEG8000 or glucosan.
7. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, it is characterized in that, the surface functional group of the polystyrene latex microballoon in described reagent R2 is amino, carboxyl, hydrazides, aldehyde radical or epoxy radicals, and the particle diameter of polystyrene latex microballoon is at 100 ~ 500nm.
8. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit, it is characterized in that, the D dimer antibody in described reagent R2 be the anti-human D dimer monoclonal antibody of rabbit, mouse-anti people D dimer monoclonal antibody one or several.
9. a kind of ion stabilizer as claimed in claim 1 and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit; it is characterized in that; protective agent 1 in described reagent R1 is EDTA; the concentration of described EDTA is 10 ~ 100mmol/L, and the protective agent 2 in described reagent R2 is bovine serum albumin(BSA).
10. the D dimer latex of surface functional group that utilizes as described in claim as arbitrary in claim 1-9 strengthens the preparation method of immunoturbidimetry detection kit, comprises the steps:
(1) preparation of reagent R1: add stabilizing agent 1, set accelerator, antiseptic 1, protective agent 1 in damping fluid 1, be uniformly mixed, obtain reagent R1;
(2) preparation of reagent R2: with mark damping fluid, polystyrene latex microspheres being diluted to mass concentration is 1%; add the EDC that mass concentration is 0.01%-0.1% again; at room temperature stirring reaction 60min; reaction terminates rear 15000rpm centrifuge washing to remove unreacted EDC; then D dimer monoclonal antibody is added; stirring reaction 4h at 37 DEG C; add stop buffer 4h cessation reaction again; by centrifugal for the reactant liquor 15000rpm obtained, washing, finally with comprising, the damping fluid 2 of stabilizing agent 2, protective agent 2, antiseptic 2 is resuspended by latex.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410736445.6A CN104360072B (en) | 2014-12-05 | 2014-12-05 | A kind of ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410736445.6A CN104360072B (en) | 2014-12-05 | 2014-12-05 | A kind of ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104360072A true CN104360072A (en) | 2015-02-18 |
| CN104360072B CN104360072B (en) | 2016-03-30 |
Family
ID=52527353
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410736445.6A Active CN104360072B (en) | 2014-12-05 | 2014-12-05 | A kind of ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104360072B (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105606803A (en) * | 2016-01-18 | 2016-05-25 | 北京九强生物技术股份有限公司 | Latex enhanced immunoturbidimetry kit for content of helicobacter pylori antibodies |
| CN105891505A (en) * | 2015-03-31 | 2016-08-24 | 北京科美生物技术有限公司 | Colloidal gold immunocolorimetry kit for detecting D-dimer (DD) and preparation method of kit |
| CN106771251A (en) * | 2017-01-10 | 2017-05-31 | 柏荣诊断产品（上海）有限公司 | Take into account the immunoglobulin G 4 hypotype IgG4 detection kits of specificity and sensitivity |
| CN106932570A (en) * | 2017-03-07 | 2017-07-07 | 中国人民解放军第二军医大学 | A kind of kit for detecting PSA PSA and its application |
| CN109557322A (en) * | 2018-12-13 | 2019-04-02 | 蓝怡科技集团股份有限公司 | A kind of ischemia modified albumin IMA calibration object and its application |
| CN110658336A (en) * | 2019-11-19 | 2020-01-07 | 北京赛科希德科技股份有限公司 | D-dimer latex immunoturbidimetry detection reagent |
| CN110865192A (en) * | 2019-11-21 | 2020-03-06 | 安徽大千生物工程有限公司 | Kit for determining GP73 based on latex enhanced immunoturbidimetry and preparation and use methods thereof |
| CN111122874A (en) * | 2019-12-31 | 2020-05-08 | 安徽大千生物工程有限公司 | Kit for determining LN based on latex enhanced immunoturbidimetry, and preparation and use methods thereof |
| 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 |
| CN112485444A (en) * | 2020-11-12 | 2021-03-12 | 安徽大千生物工程有限公司 | Kit for determining D-dimer based on latex enhanced immunoturbidimetry and preparation method thereof |
| CN115598344A (en) * | 2022-11-09 | 2023-01-13 | 山东和佰生物科技有限公司（Cn） | Detection kit for fecal pancreatic elastase 1 and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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) |
| CN103185798A (en) * | 2011-12-27 | 2013-07-03 | 苏州德沃生物技术有限公司 | Turbidimetric rapid detection kit for myocardial infarction nano-immunoenhancement and use method thereof |
| CN103558400A (en) * | 2013-11-25 | 2014-02-05 | 重庆中元生物技术有限公司 | Procalcitonin latex enhanced immunoturbidimetry detection kit |
| CN103852584A (en) * | 2014-03-28 | 2014-06-11 | 重庆中元生物技术有限公司 | Latex immune enhancement turbidimetric kit for detecting peptide C quantitatively |
-
2014
- 2014-12-05 CN CN201410736445.6A patent/CN104360072B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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) |
| CN103185798A (en) * | 2011-12-27 | 2013-07-03 | 苏州德沃生物技术有限公司 | Turbidimetric rapid detection kit for myocardial infarction nano-immunoenhancement and use method thereof |
| CN103558400A (en) * | 2013-11-25 | 2014-02-05 | 重庆中元生物技术有限公司 | Procalcitonin latex enhanced immunoturbidimetry detection kit |
| CN103852584A (en) * | 2014-03-28 | 2014-06-11 | 重庆中元生物技术有限公司 | Latex immune enhancement turbidimetric kit for detecting peptide C quantitatively |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105891505A (en) * | 2015-03-31 | 2016-08-24 | 北京科美生物技术有限公司 | Colloidal gold immunocolorimetry kit for detecting D-dimer (DD) and preparation method of kit |
| CN105606803A (en) * | 2016-01-18 | 2016-05-25 | 北京九强生物技术股份有限公司 | Latex enhanced immunoturbidimetry kit for content of helicobacter pylori antibodies |
| CN106771251A (en) * | 2017-01-10 | 2017-05-31 | 柏荣诊断产品（上海）有限公司 | Take into account the immunoglobulin G 4 hypotype IgG4 detection kits of specificity and sensitivity |
| CN106771251B (en) * | 2017-01-10 | 2019-03-15 | 柏荣诊断产品（上海）有限公司 | Take into account the immunoglobulin G 4 hypotype IgG4 detection kit of specificity and sensitivity |
| CN106932570A (en) * | 2017-03-07 | 2017-07-07 | 中国人民解放军第二军医大学 | A kind of kit for detecting PSA PSA and its application |
| CN109557322A (en) * | 2018-12-13 | 2019-04-02 | 蓝怡科技集团股份有限公司 | A kind of ischemia modified albumin IMA calibration object and its application |
| CN110658336A (en) * | 2019-11-19 | 2020-01-07 | 北京赛科希德科技股份有限公司 | D-dimer latex immunoturbidimetry detection reagent |
| CN110865192A (en) * | 2019-11-21 | 2020-03-06 | 安徽大千生物工程有限公司 | Kit for determining GP73 based on latex enhanced immunoturbidimetry and preparation and use methods thereof |
| CN111122874A (en) * | 2019-12-31 | 2020-05-08 | 安徽大千生物工程有限公司 | Kit for determining LN based on latex enhanced immunoturbidimetry, and preparation and use methods thereof |
| 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 |
| CN111999506B (en) * | 2020-08-20 | 2023-03-31 | 安徽伊普诺康生物技术股份有限公司 | D-dimer detection kit and preparation method thereof |
| CN112485444A (en) * | 2020-11-12 | 2021-03-12 | 安徽大千生物工程有限公司 | Kit for determining D-dimer based on latex enhanced immunoturbidimetry and preparation method thereof |
| CN115598344A (en) * | 2022-11-09 | 2023-01-13 | 山东和佰生物科技有限公司（Cn） | Detection kit for fecal pancreatic elastase 1 and preparation method thereof |
| CN115598344B (en) * | 2022-11-09 | 2023-04-18 | 山东和佰生物科技有限公司 | Detection kit for fecal pancreatic elastase 1 and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104360072B (en) | 2016-03-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104459139B (en) | A kind of D dimer latex of surface functional group that utilizes strengthens immunoturbidimetry detection kit | |
| CN104360072B (en) | A kind of ion stabilizer and suspension stabilizer with the use of D dimer latex strengthen immunoturbidimetry detection kit | |
| CN101819202B (en) | D-Dimer measuring kit (latex immunonephelometry method) | |
| CN103558400A (en) | Procalcitonin latex enhanced immunoturbidimetry detection kit | |
| CN101833010B (en) | Kit for detecting fibrin (ogen) degradation products (FDP) (by using emulsion immunoturbidimetry) | |
| CN103308698B (en) | Method for covalently coupling amino-containing molecules to microspheres | |
| CN1853104B (en) | Protocol and apparatus for determining heparin-induced thrombocytopenia | |
| CN101903776B (en) | Method for measuring endotoxin and reagent kit for measuring endotoxin | |
| KR101485233B1 (en) | Gel particle measurement device | |
| JP2014052388A (en) | Methods of measuring inhibition of platelet aggregation by thrombin receptor antagonists | |
| CN106405076A (en) | D-dimer detection kit and preparation method thereof | |
| CN104407156B (en) | Detect the latex enhancing immune of polysaccharide associated proteins (LBP) than turbid kit and preparation method thereof | |
| CN107942051A (en) | A kind of D dimer detection kits and its application method | |
| CN104407157B (en) | A kind of ion stabilizer and suspension stabilizer with the use of the latex enhancing immune of detection leptin than turbid kit | |
| CN104360056A (en) | Method for improving sensitivity of latex enhanced turbidimetric immunoassay | |
| CN103454193A (en) | Immunoturbidimetric kit for detecting lipoprotein (a) and preparation method thereof | |
| CN104407154A (en) | High-sensitivity latex enhanced immunoturbidimetry kit | |
| CN104535761B (en) | Hyperbranched polyglycidyl ether is modified latex microsphere and is strengthened immunoturbidimetry and application thereof | |
| CN102809654A (en) | Double-particle compounded C-reactive protein detection kit | |
| CN104459105A (en) | Latex-enhanced immunoturbidimetric assay kit for detecting leptin (LP) | |
| CN109212204A (en) | A kind of kit of latex immunoturbidimetry measurement d-dimer content | |
| CN112730848B (en) | Kit and method for detecting retinol binding protein | |
| CN104535758A (en) | Method for coupling polypeptide or protein to microsphere in covalent mode | |
| CN104391120A (en) | Latex enhanced turbidimetric immunoassay kit for detecting leptin by utilizing surface functional groups | |
| Farkas et al. | Structure, mechanical, and lytic stability of fibrin and plasma coagulum generated by Staphylocoagulase from Staphylococcus aureus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20171227 Address after: 400000 Taikang Road Dadukou District of Chongqing Bridge Industrial Park C District No. 6 Building 30 layer 1-4 Patentee after: Chongqing Zhongyuan Ji Ji Biological Technology Co., Ltd. Address before: Jiulongpo Branch Park four street 400039 Chongqing city 70-1, 70-2, J block three layer Patentee before: CHONGQING ZHONGYUAN BIOTECHNOLOGY CO., LTD. |
|
| TR01 | Transfer of patent right | ||
| CP01 | Change in the name or title of a patent holder |
Address after: Floor 1-4, building 30, no.6, Taikang Road, Zone C, Jianqiao Industrial Park, Dadukou District, Chongqing 400000 Patentee after: Zhongyuan Huiji Biotechnology Co.,Ltd. Address before: Floor 1-4, building 30, no.6, Taikang Road, Zone C, Jianqiao Industrial Park, Dadukou District, Chongqing 400000 Patentee before: CHONGQING ZHONGYUAN HUIJI BIOTECHNOLOGY Co.,Ltd. |
|
| CP01 | Change in the name or title of a patent holder |