CN107966432A - A kind of Soluble growth that measures stimulates the kit and its test method of 2 protein content of expressing gene - Google Patents
A kind of Soluble growth that measures stimulates the kit and its test method of 2 protein content of expressing gene Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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Abstract
The invention discloses the kit that Soluble growth in a kind of measure human serum using Magnetism particulate immuno chemistry luminescence method stimulates 2 albumen of expressing gene (ST2) content.Kit includes calibration object, quality-control product, anti-reagent, magnetic particle reagent, luminous substrate；The Soluble growth of its moderate resistance reagent marked by fluorescein isothiocyanate stimulates the Soluble growth of 2 albumen coated antibody of expressing gene and alkali phosphatase enzyme mark to stimulate 2 protein labeling antibody of expressing gene；Magnetic particle reagent is magnetic particle and goat-anti FITC attachments.Chemiluminescence is combined by the present invention with immune magnetic particle, provide a kind of close to homogeneous reaction system, and employ one-step method reaction pattern, so that detection sensitivity, accuracy greatly improve, detection range expands, reaction time greatly shortens, from starting to be loaded onto testing result, the time is less than 35min, hence it is evident that is faster than similar kit；And multiple samples can be measured at the same time on Full-automatic chemiluminescence apparatus, realize that Soluble growth stimulates the rapid measure of high throughput of 2 albumen of expressing gene, accuracy is high, and high specificity, accuracy and detection efficiency are enhanced.
Description
Technical field
The present invention relates to technological field of biochemistry, and in particular to one kind is using in Magnetism particulate immuno chemistry luminescence method measure human body
Soluble growth stimulates the kit of 2 albumen of expressing gene (ST2) content.
Background technology
ST2 as emerging biological marker, have biologic variability is low, stability is high, from gender, the age, race,
The features such as renal function disturbs, plays an important role, the ST2 in serum in the diagnosis, treatment and risk profile of patients with heart failure
The horizontal order of severity with heart failure is proportionate, and from the influence of the indexs such as age, weight, plays and hinders in angiocardiopathy
The anti-myocardial hypertrophy of disconnected IL-33, resisting myocardial fibrillation, study of anti-atherogenic effect.In addition in acute coronary artery syndrome
It may also be played an important role in occurrence and development, marker of inflammation and the heart as reflection coronary artery vulnerable plaque degree of stability
Effective biological markers of vascular diseases poor prognosis.It is alternatively arranged as potentially representing effective biology of cardiac mechanical excess load
Marker is learned, and independently of other dlinial prediction factors.
The Soluble growth being currently known stimulates 2 protein assay method enzyme linked immunosorbent assay (ELISA) of expressing gene.Enzyme
There are detection time is long, complicated, poor repeatability, is unsuitable for emergency treatment and need that clinical patient diagnoses in time for connection immunoabsorption
Will.
The content of the invention
The defects of the technical problem to be solved in the present invention is the prior art is overcome, there is provided a kind of high use magnetic of accuracy is micro-
Soluble growth stimulates the kit of 2 albumen of expressing gene (ST2) content in grain chemiluminescence determination human serum.
In order to solve the above technical problem, the present invention provides following technical solution：
A kind of Soluble growth that measures stimulates the kit of 2 protein content of expressing gene, including calibration object, quality-control product, anti-
Reagent, magnetic particle reagent, luminous substrate；
The magnetic particle reagent is that anti-fluorescein isothiocynate antibody and the coupling of carboxyl magnetic bead are made, the luminous substrate
It is that ALPS is dissolved in luminous substrate buffer solution to be made；
Calibration object, quality-control product, anti-reagent, magnetic particle reagent, luminous substrate are prepared respectively；Soluble growth is stimulated and is expressed
2 albumen calibration object of gene, Soluble growth stimulate 2 albumen quality-control product of expressing gene, anti-reagent, magnetic particle reagent, luminous substrate,
It is separately applied in packing container, obtaining Soluble growth stimulates the quantitative determination reagent kit of 2 albumen of expressing gene；
(1), the preparation of the calibration object, quality-control product, comprises the following steps：
2 proteantigen of expressing gene is stimulated with calibration object buffer solution dissolving Soluble growth, prepares the calibration of anti-reagent
Product and quality-control product；Wherein, the calibration object buffer solution is by adding 0.01g~0.05g in the newborn bovine serum of 1L
The neomycinsulphate of tetracycline and 0.1g~0.5g, is prepared after being completely dissolved by 0.22 μm of filter membrane processing；
(2), the preparation of the anti-reagent：
1) preparation of anti-reagent buffer：
By the Na of 10g~20g2PO3H·12H2O, the NaPO of 1~2g3H2·12H2O, the sheep serum of 1g~5g, 3g~
The newborn bovine serum of 10g, the horse serum of 1g~5g are added in 1L purified waters, are stirred well to and are completely dissolved, and are adjusted with 4M HCl
PH to 5~6, is made the anti-reagent buffer；
2) fluorescein isothiocynate stimulates 2 protein antibodies of expressing gene to be coupled with Soluble growth, and it is glimmering to obtain isothiocyanic acid
The Soluble growth of light element mark stimulates 2 albumen coated antibody of expressing gene：
Fluorescein isothiocynate is configured to the fluorescein isothiocynate that concentration is 1.0~5.0mg/mL with buffer solution first
Solution, then stimulates 2 protein antibodies of expressing gene according to Soluble growth：Fluorescein isothiocynate solution=1:1.1~1:1.5
Mass ratio the two is transferred in Brown Glass Brown glass bottles and jars only, fully mix；The carbonate buffer that pH is 8~9 is fully used after reaction
Liquid balances, and then isolating and purifying to obtain the Soluble growth of marked by fluorescein isothiocyanate using gel chromatography stimulates expressing gene
2 albumen coated antibodies；
3) alkaline phosphatase stimulates 2 protein antibodies of expressing gene to be coupled with Soluble growth, obtains alkali phosphatase enzyme mark
Soluble growth stimulate 2 protein antibodies of expressing gene：
Alkaline phosphatase is configured to the alkaline phosphatase enzyme solutions that concentration is 1.0~5.0mg/mL with buffer solution first, can
2 protein antibodies of dissolubility growth stimulation expressing gene and alkaline phosphatase according to molar ratio are after reactive group is activated respectively respectively
Alkaline phosphatase：Soluble growth stimulates 2 albumen=1 of expressing gene:1~1:3 reaction is more abundant than under the catalysis of catalyst
Mix and carry out coupling reaction, fully after reaction, balanced using the tris buffer solutions that pH is 8~9, it is big that gel column carries out different molecular
Small pieces degree isolates and purifies, and obtains the Soluble growth of alkali phosphatase enzyme mark and stimulates 2 protein labeling antibody of expressing gene；
Stimulate 2 albumen of expressing gene coating anti-the Soluble growth of the marked by fluorescein isothiocyanate obtained in step 2)
The Soluble growth of the alkali phosphatase enzyme mark obtained in body and step 3) stimulates the addition of 2 protein labeling antibody of expressing gene to contain
In the phosphate buffer of surfactant, the anti-reagent is obtained after being sufficiently stirred；Preferably, the surface in the step 3)
Activating agent is Tween 20, the one or more in Triton X-100, Bronidox, and the additive amount of surfactant is
0.01%~0.5%.
Further, which further includes cleaning solution, the collocation method of the cleaning solution will by 160gNaCl, 4gKCl,
24.2g trishydroxymethylaminomethanes, 1mL polysorbas20s are dissolved in 900ml distilled waters, are adjusted to PH7.4 with HCL, are determined with distilled water
Hold to 1000ml.With 15 times of dilutions of distilled water during use.
Further, the magnetic particle reagent is prepared according to following steps：
1) the carboxyl magnetic bead concentrate after abundant mix is put into reaction bulb, which is placed in magnetic field to 15~
20min, sucks supernatant after carboxyl magnetic bead all sedimentation, is added into reaction bulb equivalent to carboxyl magnetic bead volume 2 in reaction bulb
~5 times of magnetic particle buffer solution, 20~30min of concussion cleaning；Reaction bulb is placed in magnetic field after 15~20min again and is sucked
Clearly；Repeated washing carboxyl magnetic bead 3 times；Finally by carboxyl magnetic bead solution constant volume to 10~50mg/mL, mix stand-by；
2) coupled reaction according to quality than carboxyl magnetic bead solution：Anti- fluorescein isothiocynate antibody=100:1 ratio exists
Anti- fluorescein isothiocynate antibody is added in carboxyl magnetic bead solution obtained by step 1), keeps mixing state anti-in 2~8 DEG C
Answer 18 it is small when；
3) reaction bulb places 15min in magnetic field, is washed 3 times with magnetic particle buffer solution after the sedimentation of carboxyl magnetic bead, then fixed
Hold to 10mg/mL, 2~8 DEG C of preservations, are made required stand-by magnetic particle reagent.Preferably, the configuration side of the magnetic particle buffer solution
Method is by the Tris of 12.12~15.26mg, the sodium chloride of 5.82~8.58mg, and it is pure that the Methyl cellulose ether of 50~60g is added to 1L
Change in water, be stirred well to and be completely dissolved to obtain the final product.
Further, the luminous substrate is fully molten with luminous substrate buffer solution of 4~10 times equivalent to ALPS volumes
Solution ALPS is prepared；The collocation method of the luminous substrate buffer solution is by Tris, 5.82g of 12.12g~121.14g
Sodium chloride, the lucigenin of 0.03g be added in 1L purified waters, be stirred well to and be completely dissolved, be with salt acid for adjusting pH to 9.5
.
The method that 2 protein content of expressing gene is stimulated using the kit measurement Soluble growth, is comprised the following steps：
1) three test tubes are taken to add 100 μ L calibration objects, 100 μ L quality-control products, 100 μ L samples to be tested respectively；
2) the 60 anti-reagents of μ L are added in every test tube, with covered rearing with plastic film test tube, test tube 30s is gently vibrated, is placed in 37
Water-bath 15min at DEG C；
3) 30 μ L magnetic particle reagents are added in every test tube, with covered rearing with plastic film test tube, test tube 30s is gently vibrated, puts
Water-bath 5min at 37 DEG C；
4) test tube is precipitated to 3min on magnetic separator, test tube and magnetic separator is slowly reversed, pours out supernatant；Falling
The test tube turned is placed on filter paper together with magnetic separator, flops magnetic separator bottom to remove all liquid being sticked on tube wall
Drop；
5) 300 μ L cleaning solutions are added in every test tube, with covered rearing with plastic film test tube, test tube 30s are gently vibrated, after mixing
Test tube and magnetic separator are slowly reversed, pours out supernatant, the test tube of reversing together with magnetic separator, is placed on filter paper,
Separator bottom is firmly flopped to remove all drops being sticked on tube wall；
6) repeat step 5) once；
7) 200 μ L luminous substrates are added in every test tube, vibration mixes 3s, and luminous intensity is detected with Chemiluminescence Apparatus.
The magnetic particle reagent of the present invention is magnetic particle and goat-anti FITC attachments, the buffer solution containing BSA.
The calibration object of the present invention is to the addition of not same amount Soluble growth respectively to stimulate containing for 2 proteantigen of expressing gene
The buffer solution of BSA.
The present invention technical principle be：The ST2 antibody and alkaline phosphatase (AP) of fluorescein isothiocynate (FITC) mark
ST2 in the ST2 pairing antibody and sample, calibration object or quality-control product of mark combines to form " sandwich " compound.Then add
The magnetic particle of anti-FITC antibody is connected with, antigenantibody complex knot is made by the specific binding of anti-FITC antibody and FITC
Close on magnetic particle.Under the action of externally-applied magnetic field, the compound that immune response is formed is separated with other uncombined materials,
After cleaning compound, enzyme-catalyzed chemical luminescence substrate is added.Substrate, by catalytic pyrolysis, forms unstable excitation state under enzyme effect
Intermediate, photon is sent when excitation state intermediate returns to ground state, forms luminescence-producing reaction, you can is detected using Chemiluminescence Apparatus anti-
The luminous intensity answered.In detection range, luminous intensity is directly proportional to the content of the ST2 in sample, uses four parameters of improvement
Logistic equation models can calculate ST2 concentration in sample.
The beneficial effect that is reached of the present invention is：
1st, chemiluminescence is combined by the kit with immune magnetic particle, there is provided a kind of close to homogeneous reactant
System, and employs one-step method reaction pattern so that detection sensitivity, accuracy greatly improve, detection range expands, during reaction
Between greatly shorten, from starting to be loaded onto testing result, the time is less than 35min, hence it is evident that is faster than similar kit；
2nd, a kind of new FITC antibody and magnetic particle coupling method have been invented, this method coupling efficiency is high, is firmly combined with, and
Process stabilizing, while enhancing product performance, greatly reduces product cost.
3rd, kit moderate resistance reagent, magnetic particle reagent, calibration object, quality-control product, luminous substrate liquid and concentration washing lotion are these
Optimization formula under reaction system, the phase is imitated in the use to the kit and detection performance provides powerful guarantee.
4th, the present invention can measure multiple samples at the same time on Full-automatic chemiluminescence apparatus, realize Soluble growth stimulation table
Up to the rapid measure of high throughput of 2 albumen of gene, accuracy is high, and high specificity, accuracy and detection efficiency have larger carry
It is high.
Brief description of the drawings
Attached drawing is used for providing a further understanding of the present invention, and a part for constitution instruction, the reality with the present invention
Apply example to be used to explain the present invention together, be not construed as limiting the invention.In the accompanying drawings：
Fig. 1 is the correlation of the kit and other commercial reagent box detection clinical serums of the present invention；Wherein abscissa is
The testing result of other commercial reagent boxes, ordinate are the testing result of kit of the present invention.
Embodiment
The preferred embodiment of the present invention is illustrated below, it will be appreciated that preferred embodiment described herein is only used
In the description and interpretation present invention, it is not intended to limit the present invention.
The operation sequence of kit test sample using the present invention is as follows：
1st, sample collection
Serum (sample of significant hemolysis or piarhemia cannot be used for measuring), the sample after collection are collected using correct medical technology
This room temperature place may not exceed 8 it is small when；If when 8 is small, sample need to be positioned in 2-8 DEG C of refrigerator by interior detection；If need
72 it is small when more than preserve or transport, then should freeze in less than -20 DEG C, avoid multigelation.Room temperature is returned to before use, is gently shaken
It is dynamic to mix.
2nd, prepare before experiment
1. one bottle of washing lotion is taken to carry out 15 times of dilutions with distilled water；
2. insulating box or water-bath pot temperature are adjusted to 37 DEG C, used after temperature stabilization；
3. by magnetic particle suspension fully mix to be visible by naked eyes precipitation.
3 experimental methods
1. taking out a certain amount of reaction vessel (flat based tubes), number.100ul calibration objects/matter is added according to requirement of experiment
Control product/clinical sample；
2. it is separately added into anti-reagent 60ul per hole；
3. solution in reaction vessel is uniformly mixed, 37 DEG C incubate 15 minutes；
4. shaking up magnetic particle suspension, 30ul is separately added into per hole；
5. solution in reaction vessel is uniformly mixed, 37 DEG C incubate 5 minutes；
6. reaction vessel is taken out, using Magneto separate and washing facility, by the wash liquid 3 times of magnetic particle in reaction vessel；
7. going washing lotion after the completion of washing, add luminous substrate liquid 200ul per hole, shake；
8. chemiluminescence detector device detects luminous intensity；
9. using four parameter fitting modes, using calibration object concentration value as X-axis, using calibration object luminous intensity values as Y-axis, establish
Calibration curve.Corresponding concentration value is back-calculated according to the luminous intensity values of sample to be tested.
Kit of the present invention is identified according to methodology, can reach following index：
Standard curve is linear：R ＞ 0.9900.
Minimum detection limit：≤1.31pg/ml.
Accuracy：TIANZHU XINGNAO Capsul 85%-115%.
Repeatability：Coefficient of variation CV≤8%.
Difference between batch：The coefficient of variation≤15%.
Linear dilution：R is more than 0.9900.
2 protein detection kit A of expressing gene is stimulated to contrast 100 parts of serum sample measured values and commercially available Soluble growth,
Compare the correlation that kit of the present invention stimulates 2 protein reagent box testing result of expressing gene with commercially available Soluble growth, as a result
As shown in Figure 1.
Finally it should be noted that：The foregoing is only a preferred embodiment of the present invention, is not intended to limit the invention,
Although the present invention is described in detail with reference to the foregoing embodiments, for those skilled in the art, it still may be used
To modify to the technical solution described in foregoing embodiments, or equivalent substitution is carried out to which part technical characteristic.
Within the spirit and principles of the invention, any modification, equivalent replacement, improvement and so on, should be included in the present invention's
Within protection domain.
Claims (7)
1. it is a kind of measure Soluble growth stimulate 2 protein content of expressing gene kit, it is characterised in that including calibration object,
Quality-control product, anti-reagent, magnetic particle reagent, luminous substrate；
The magnetic particle reagent be by anti-fluorescein isothiocynate antibody and carboxyl magnetic bead coupling be made, the luminous substrate be by
ALPS is dissolved in luminous substrate buffer solution and is made；
Calibration object, quality-control product, anti-reagent, magnetic particle reagent, luminous substrate are prepared respectively；By calibration object, quality-control product, anti-reagent,
Magnetic particle reagent, luminous substrate, are separately applied in packing container, and obtaining Soluble growth stimulates determining for 2 albumen of expressing gene
Measure assay kit；
(1), the preparation of the calibration object, quality-control product, comprises the following steps：
With calibration object buffer solution dissolving Soluble growth stimulate 2 proteantigen of expressing gene, prepare anti-reagent calibration object and
Quality-control product；Wherein, the calibration object buffer solution is the Fourth Ring by adding 0.01g~0.05g in the newborn bovine serum of 1L
The neomycinsulphate of element and 0.1g~0.5g, is prepared after being completely dissolved by 0.22 μm of filter membrane processing；
(2), the preparation of the anti-reagent：
1) preparation of anti-reagent buffer：
By the Na of 10g~20g2PO3H·12H2O, the NaPO of 1~2g3H2·12H2O, the sheep serum of 1g~5g, 3g~10g
Newborn bovine serum, the horse serum of 1g~5g are added in 1L purified waters, are stirred well to and are completely dissolved, and pH to 5 is adjusted with 4M HCl
~6, the anti-reagent buffer is made；
2) fluorescein isothiocynate stimulates 2 protein antibodies of expressing gene to be coupled with Soluble growth, obtains fluorescein isothiocynate
The Soluble growth of mark stimulates 2 albumen coated antibody of expressing gene：
First with buffer solution by fluorescein isothiocynate be configured to concentration be 1.0~5.0mg/mL fluorescein isothiocynate it is molten
Liquid, then stimulates 2 protein antibodies of expressing gene according to Soluble growth：Fluorescein isothiocynate solution=1:1.1~1:1.5
The two is transferred in Brown Glass Brown glass bottles and jars only by mass ratio, fully mixes；The carbonate buffer solution that pH is 8~9 is fully used after reaction
Balance, then isolating and purifying to obtain the Soluble growth of marked by fluorescein isothiocyanate using gel chromatography stimulates expressing gene 2
Albumen coated antibody；
3) alkaline phosphatase and Soluble growth stimulate 2 protein antibodies of expressing gene to be coupled, and obtain alkali phosphatase enzyme mark can
2 protein antibodies of dissolubility growth stimulation expressing gene：
Alkaline phosphatase is configured to the alkaline phosphatase enzyme solutions that concentration is 1.0~5.0mg/mL with buffer solution first, it is soluble
2 protein antibodies of growth stimulation expressing gene and alkaline phosphatase reactive group is activated respectively respectively after according to molar ratio for alkalescence
Phosphatase：Soluble growth stimulates 2 albumen=1 of expressing gene:1~1:3 reaction ratio fully mixes under the catalysis of catalyst
Coupling reaction is carried out, fully after reaction, is balanced using the tris buffer solutions that pH is 8~9, gel column carries out different molecular big or small slice
Degree isolates and purifies, and obtains the Soluble growth of alkali phosphatase enzyme mark and stimulates 2 protein labeling antibody of expressing gene；
By the Soluble growth of the marked by fluorescein isothiocyanate obtained in step 2) stimulate 2 albumen coated antibody of expressing gene and
The Soluble growth of the alkali phosphatase enzyme mark obtained in step 3) stimulates the addition of 2 protein labeling antibody of expressing gene to contain surface
In the phosphate buffer of activating agent, the anti-reagent is obtained after being sufficiently stirred.
2. measure Soluble growth according to claim 1 stimulates the kit of 2 protein content of expressing gene, its feature exists
In the kit further includes cleaning solution, and the collocation method of the cleaning solution will be by 160gNaCl, 4gKCl, 24.2g trihydroxy methyl
Aminomethane, 1mL polysorbas20s are dissolved in 900ml distilled waters, are adjusted with HCL to PH7.4, and 1000ml is settled to distilled water.
3. measuring Soluble growth according to 1 or 2 any one of them of claim stimulates the reagent of 2 protein content of expressing gene
Box, it is characterised in that the surfactant in the step 3) is Tween 20, one kind in TritonX-100, Bronidox
Or it is a variety of, the additive amount of surfactant is 0.01%~0.5%.
4. measuring Soluble growth according to 1 or 2 any one of them of claim stimulates the reagent of 2 protein content of expressing gene
Box, it is characterised in that the magnetic particle reagent is prepared according to following steps：
1) the carboxyl magnetic bead concentrate after abundant mix is put into reaction bulb, which is placed in magnetic field to 15~
20min, sucks supernatant after carboxyl magnetic bead all sedimentation, is added into reaction bulb equivalent to carboxyl magnetic bead volume 2 in reaction bulb
~5 times of magnetic particle buffer solution, 20~30min of concussion cleaning；Reaction bulb is placed in magnetic field after 15~20min again and is sucked
Clearly；Repeated washing carboxyl magnetic bead 3 times；Finally by carboxyl magnetic bead solution constant volume to 10~50mg/mL, mix stand-by；
2) coupled reaction：According to quality than carboxyl magnetic bead solution：Anti- fluorescein isothiocynate antibody=100:1 ratio is in step
1) anti-fluorescein isothiocynate antibody is added in the carboxyl magnetic bead solution obtained by, keeps mixing state response 18 in 2~8 DEG C
Hour；
3) reaction bulb places 15min in magnetic field, is washed 3 times with magnetic particle buffer solution after the sedimentation of carboxyl magnetic bead, is then settled to
10mg/mL, 2~8 DEG C of preservations, are made required stand-by magnetic particle reagent.
5. measuring Soluble growth according to claim 4 any one of them stimulates the kit of 2 protein content of expressing gene, its
It is characterized in that, the collocation method of the magnetic particle buffer solution is by the Tris of 12.12~15.26mg, the chlorine of 5.82~8.58mg
Change sodium, the Methyl cellulose ether of 50~60g is added in 1L purified waters, is stirred well to and is completely dissolved to obtain the final product.
6. measure Soluble growth according to claim 1 stimulates the kit of 2 protein content of expressing gene, its feature exists
In：The luminous substrate is fully to dissolve ALPS with luminous substrate buffer solution of 4~10 times equivalent to ALPS volumes to be prepared
's；The collocation method of the luminous substrate buffer solution is by the sodium chloride of Tris, 5.82g of 12.12g~121.14g, 0.03g
Lucigenin be added in 1L purified waters, be stirred well to and be completely dissolved, with salt acid for adjusting pH to 9.5 to obtain the final product.
7. stimulate 2 protein content of expressing gene using 1 or 2 any one of them kit measurement Soluble growth of claim
Method, it is characterised in that this method comprises the following steps：
1) three test tubes are taken to add 100 μ L calibration objects, 100 μ L quality-control products, 100 μ L samples to be tested respectively；
2) the 60 anti-reagents of μ L are added in every test tube, with covered rearing with plastic film test tube, test tube 30s is gently vibrated, is placed at 37 DEG C
Water-bath 15min；
3) 30 μ L magnetic particle reagents are added in every test tube, with covered rearing with plastic film test tube, test tube 30s is gently vibrated, puts 37 DEG C
Lower water-bath 5min；
4) test tube is precipitated to 3min on magnetic separator, test tube and magnetic separator is slowly reversed, pours out supernatant；Reversing
Test tube is placed on filter paper together with magnetic separator, flops magnetic separator bottom to remove all drops being sticked on tube wall；
5) 300 μ L cleaning solutions are added in every test tube, with covered rearing with plastic film test tube, gently vibrate test tube 30s, after mixing slowly
Reversing test tube and magnetic separator, pour out supernatant, the test tube of reversing together with magnetic separator, be placed on filter paper, firmly
Separator bottom is flopped to remove all drops being sticked on tube wall；
6) repeat step 5) once；
7) 200 μ L luminous substrates are added in every test tube, vibration mixes 3s, and luminous intensity is detected with Chemiluminescence Apparatus.
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Cited By (5)
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| CN110208549A (en) * | 2019-07-01 | 2019-09-06 | 北京利德曼生化股份有限公司 | A kind of Soluble growth stimulation 2 albumen sST2 luminescence reagent box of expressing gene |
| CN111337664A (en) * | 2020-02-27 | 2020-06-26 | 江苏泽成生物技术有限公司 | Neutrophilic granulocyte gelatin-related lipocalin detection kit and use method thereof |
| CN113588967A (en) * | 2021-08-03 | 2021-11-02 | 湖南永和阳光生物科技股份有限公司 | Magnetic particle chemiluminescence detection kit for growth stimulation expression gene 2 protein |
| CN113702647A (en) * | 2021-08-31 | 2021-11-26 | 普十生物科技(北京)有限公司 | Human growth differentiation factor 15 instant detection kit, preparation method and application thereof |
| CN113777324A (en) * | 2021-09-10 | 2021-12-10 | 普十生物科技(北京)有限公司 | Growth stimulation expression factor 2 instant detection kit, preparation method and application thereof |
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