CN108226515A - A kind of kit and its test method for measuring sex hormone binding globulin content - Google Patents
A kind of kit and its test method for measuring sex hormone binding globulin content Download PDFInfo
- Publication number
- CN108226515A CN108226515A CN201711217126.4A CN201711217126A CN108226515A CN 108226515 A CN108226515 A CN 108226515A CN 201711217126 A CN201711217126 A CN 201711217126A CN 108226515 A CN108226515 A CN 108226515A
- Authority
- CN
- China
- Prior art keywords
- sex hormone
- hormone binding
- binding globulin
- reagent
- test tube
- 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.)
- Pending
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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
-
- 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
-
- 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
- G01N33/543—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
Abstract
The invention discloses a kind of kits that human serum Sex Hormones binding protein (SHBG) content is measured using Magnetism particulate immuno chemistry luminescence method.Kit includes calibration object, quality-control product, anti-reagent, magnetic particle reagent, luminous substrate；The sex hormone binding globulin coated antibody of its moderate resistance reagent marked by fluorescein isothiocyanate and the sex hormone binding globulin labelled antibody of alkali phosphatase enzyme mark；Magnetic particle reagent is magnetic particle and goat-anti FITC attachments.Chemiluminescence is combined by the present invention with immune magnetic particle, it provides 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 25min, hence it is evident that is faster than similar kit；And multiple samples can be measured simultaneously on Full-automatic chemiluminescence apparatus, realize the rapid measure of high throughput of sex hormone binding globulin, 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 a kind of to be measured in human body using Magnetism particulate immuno chemistry luminescence method
The kit of sex hormone binding globulin (SHBG) content.
Background technology
A kind of glycoprotein for being responsible for testosterone and estradiol progress blood-transmitted of sex hormone binding globulin (SHBG).SHBG is
It is synthesized in liver, affinity is very high when it is combined with testosterone and 5- dihydro-testosterones, and affinity summary when being combined with estradiol
It is low.In mating type testosterone, 44%-60% is combined with SHBG；Estrogen about 95% is tied in blood circulation with special SHBG phases
It closes.
Female hirsutism and manlike SHBG contents are shown in clinical symptoms in female hirsutism and manlike disease,
Only the 50% of normal value, and free T contents then almost increase by 90%.Therefore SHBG measure can be used as examining in this kind of disease
Severed finger mark, and can be as the foundation for weighing therapeutic effect.Stein-Leventhal syndrome (PCOS), obesity, thyroid function are low
Lower SHBG levels decline.It can see in the patient's body for having liver diseases (hepatic sclerosis, chronic hepatitis, fatty liver) raised
SHBG is horizontal, and oral contraceptive and anti-epileptic will can result in the raising of SHBG levels.
The sex hormone binding globulin assay method being currently known has radioimmunology (RIA), enzyme linked immunosorbent assay
(ELISA), latex-enhanced turbidimetry etc..Radioimmunology complex steps, reagent price is expensive, need to use mating instrument and deposit
In radioactive pollution.Enzyme linked immunosorbent assay there are detection time it is long, complicated for operation, poor repeatability, be unsuitable for emergency treatment and clinic
The needs that patient diagnoses in time.Latex enhancing immune turbidimetry is easy to operate, quick, but sensitivity is low, low value poor repeatability.
Invention content
The technical problem to be solved by the present invention is to overcome the defects of the prior art, providing, a kind of high use magnetic of accuracy is micro-
The kit of grain chemiluminescence determination human serum Sex Hormones binding protein (SHBG) content.
In order to solve the above technical problem, the present invention provides following technical solutions：
A kind of kit for measuring sex hormone binding globulin content, including calibration object, quality-control product, anti-reagent, magnetic particle examination
Agent, 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；By calibration object, quality-control product, anti-examination
Agent, magnetic particle reagent, luminous substrate, are separately applied in packing container, obtain the quantitative determination reagent of sex hormone binding globulin
Box；
(1), the preparation of the calibration object, quality-control product, includes the following steps：
Sex hormone binding globulin antigen is dissolved with calibration object buffer solution, prepares the calibration object and quality-control product of anti-reagent；Its
In, the calibration object buffer solution be by the newborn bovine serum of 1L add in 0.01g~0.05g tetracycline and 0.1g~
The neomycinsulphate of 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
The anti-reagent buffer is made in pH to 5~6；
2) fluorescein isothiocynate and sex hormone binding globulin antibody coupling, the property for obtaining marked by fluorescein isothiocyanate swash
Plain binding protein coated antibody：
Fluorescein isothiocynate is configured to the fluorescein isothiocynate of a concentration of 1.0~5.0mg/mL with buffer solution first
Solution, then according to sex hormone binding globulin antibody：Fluorescein isothiocynate solution=1:1.1~1:1.5 mass ratio is by two
Person is transferred in Brown Glass Brown glass bottles and jars only, abundant mixing；It is fully balanced using the carbonate buffer solution that pH is 8~9 after reaction, then made
It is isolated and purified to obtain the sex hormone binding globulin coated antibody of marked by fluorescein isothiocyanate with gel chromatography；
3) alkaline phosphatase and sex hormone binding globulin antibody coupling obtain the sex hormone combination egg of alkali phosphatase enzyme mark
Bai Kangti：
Alkaline phosphatase is configured to the alkaline phosphatase enzyme solutions of a concentration of 1.0~5.0mg/mL, property with buffer solution first
Hormonebinding protein antibody and alkaline phosphatase reactive group is activated respectively respectively after according to molar ratio be alkaline phosphatase：Property
Hormonebinding protein=1:1~1:3 reaction carries out coupling reaction than mixing abundant under the catalysis of catalyst, fully reacts
Afterwards, it is balanced using the tris buffer solutions that pH is 8~9, gel column carries out isolating and purifying for different molecular big or small slice degree, obtains alkalinity
The sex hormone binding globulin labelled antibody of phosphatase enzyme mark；
It will be in the sex hormone binding globulin coated antibody and step 3) of the marked by fluorescein isothiocyanate that obtained in step 2)
The sex hormone binding globulin labelled antibody of the alkali phosphatase enzyme mark of acquisition adds in the phosphate buffer containing surfactant
In, the anti-reagent is obtained after being sufficiently stirred；Preferably, the surfactant in the step 3) is Tween 20, Triton
One or more in X-100, Bronidox, the additive amount of surfactant is 0.01%~0.5%.
Further, which further includes cleaning solution, the configuration 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 mixing is put into reaction bulb, which is placed in magnetic field to 15~
20min, supernatant is sucked after carboxyl magnetic bead all sedimentation, is added in into reaction bulb and is 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, mixing is for use；
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
It answers 18 hours；
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, required for use magnetic particle reagent is made.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 the luminous substrate buffer solution for being equivalent to 4~10 times of ALPS volumes
Solution ALPS is prepared；The configuration method of the luminous substrate buffer solution is by Tris, 5.82g of 12.12g~121.14g
Sodium chloride, 0.03g lucigenin be added in 1L purified waters, be stirred well to and be completely dissolved, be with salt acid for adjusting pH to 9.5
.
Using the method for the kit measurement sex hormone binding globulin content, include the following steps：
1) three test tubes is taken to add 20 μ L calibration objects, 20 μ L quality-control products, 20 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 5min 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 on magnetic separator is precipitated to 3min, 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, gently vibrate test tube 30s, 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) it is primary to repeat step 5)；
7) 200 μ L luminous substrates are added in every test tube, vibrate mixing 3s, 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 the buffer solution containing BSA for being added to not same amount sex hormone binding globulin antigen respectively.
The present invention technical principle be：The SHBG antibody of fluorescein isothiocynate (FITC) label and alkaline phosphatase (AP)
SHBG in the SHBG pairing antibody and sample, calibration object or quality-control product of label, which is combined, forms " sandwich " compound.Then add
Enter to be connected with the magnetic particle of anti-FITC antibody, antigenantibody complex is made by the specific binding of anti-FITC antibody and FITC
It is incorporated on magnetic particle.Under the action of externally-applied magnetic field, by the compound that immune response is formed and unbonded other substances point
From, after cleaning compound, addition enzyme-catalyzed chemical luminescence substrate.Substrate by catalytic pyrolysis, is formed and unstable is swashed under enzyme effect
State intermediate is sent out, photon is sent out when excitation state intermediate returns to ground state, forms luminescence-producing reaction, you can examine using Chemiluminescence Apparatus
Survey the luminous intensity of reaction.In detection range, luminous intensity is directly proportional to the content of the SHBG in sample, uses the four of improvement
Parameter Logistic equation models can calculate SHBG concentration in sample.
The advantageous effect that is reached of the present invention is：
1st, chemiluminescence is combined by the kit with immune magnetic particle, 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 25min, 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 imitates the phase to the use of the kit and detection performance provides powerful guarantee.
4th, the present invention can measure multiple samples simultaneously on Full-automatic chemiluminescence apparatus, realize sex hormone binding globulin
High-throughput rapid measure, accuracy is high, and high specificity, accuracy and detection efficiency are enhanced.
Description of the drawings
Attached drawing is used to provide further understanding of the present invention, and a part for constitution instruction, the reality with the present invention
Example is applied together for explaining the present invention, is 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.
Specific embodiment
The preferred embodiment of the present invention is illustrated below, it should be understood 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 may not exceed 8 hours being placed at room temperature for；Sample need to be positioned in 2-8 DEG C of refrigerator if detected not in 8 hours；If it needs
It preserves or transports within 72 hours or more, then should freeze in -20 DEG C hereinafter, avoiding multigelation.Room temperature is returned to before use, is gently shaken
Dynamic mixing.
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 the abundant mixing of magnetic particle suspension to be visible by naked eyes precipitation.
3 experimental methods
1. taking out a certain amount of reaction vessel (flat based tubes), number.30ul calibration objects/Quality Control is added according to requirement of experiment
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 5 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 according to methodology is identified, can reach following index：
Standard curve is linear：R ＞ 0.9900.
Minimum detection limit：≤0.06ng/ml.
Accuracy：Relative deviation is in the range of ± 10%.
Repeatability：Coefficient of variation CV≤8%.
Difference between batch：The coefficient of variation≤15%.
Linear dilution：R is more than 0.9900.
100 parts of serum sample measured values and commercially available sex hormone binding globulin detection kit A are compared, reagent more of the present invention
Box and the correlation of commercially available sex hormone binding globulin kit testing result, the results are 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 restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
To modify to the technical solution recorded in foregoing embodiments or carry out equivalent replacement to which part technical characteristic.
All within the spirits and principles of the present 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 sex hormone binding globulin content kit, which is characterized 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, obtain the quantitative determination reagent kit of sex hormone binding globulin；
(1), the preparation of the calibration object, quality-control product, includes the following steps：
Sex hormone binding globulin antigen is dissolved with calibration object buffer solution, prepares the calibration object and quality-control product of anti-reagent；Wherein, institute
It is the tetracycline and 0.1g~0.5g by adding in 0.01g~0.05g in the newborn bovine serum of 1L to state calibration object buffer solution
Neomycinsulphate, be prepared after being completely dissolved by the processing of 0.22 μm of filter membrane；
(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, 1g~
In the sheep serum of 5g, the newborn bovine serum of 3g~10g, the horse serum addition 1L purified waters of 1g~5g, it is stirred well to completely
Dissolving adjusts pH to 5~6 with 4M HCl, the anti-reagent buffer is made；
2) fluorescein isothiocynate and sex hormone binding globulin antibody coupling obtain the sex hormone knot of marked by fluorescein isothiocyanate
Hop protein coated antibody：
The fluorescein isothiocynate that fluorescein isothiocynate is configured to a concentration of 1.0~5.0mg/mL with buffer solution first is molten
Liquid, then according to sex hormone binding globulin antibody：Fluorescein isothiocynate solution=1:1.1~1:1.5 mass ratio is by the two
It is transferred in Brown Glass Brown glass bottles and jars only, abundant mixing；It is fully balanced using the carbonate buffer solution that pH is 8~9 after reaction, then used
Gel chromatography isolates and purifies to obtain the sex hormone binding globulin coated antibody of marked by fluorescein isothiocyanate；
3) alkaline phosphatase and sex hormone binding globulin antibody coupling, the sex hormone binding globulin for obtaining alkali phosphatase enzyme mark resist
Body：
Alkaline phosphatase is configured to the alkaline phosphatase enzyme solutions of a concentration of 1.0~5.0mg/mL, sex hormone with buffer solution first
Binding protein antibody and alkaline phosphatase reactive group is activated respectively respectively after according to molar ratio be alkaline phosphatase：Sex hormone
Binding protein=1:1~1:3 reaction carries out coupling reaction than mixing abundant under the catalysis of catalyst, fully after reaction, makes
It is balanced with the tris buffer solutions that pH is 8~9, gel column carries out isolating and purifying for different molecular big or small slice degree, obtains alkaline phosphatase
The sex hormone binding globulin labelled antibody of enzyme label；
It will be obtained in the sex hormone binding globulin coated antibody of the marked by fluorescein isothiocyanate obtained in step 2) and step 3)
Alkali phosphatase enzyme mark sex hormone binding globulin labelled antibody add in the phosphate buffer containing surfactant in, fill
The anti-reagent is obtained after dividing stirring.
2. the kit according to claim 1 for measuring sex hormone binding globulin content, which is characterized in that the kit is also
Including cleaning solution, the configuration method of the cleaning solution will spit 160gNaCl, 4gKCl, 24.2g trishydroxymethylaminomethane, 1mL
Temperature 20 is dissolved in 900ml distilled waters, is adjusted with HCL to PH7.4,1000ml is settled to distilled water.
3. the kit of sex hormone binding globulin content is measured according to claims 1 or 2 any one of them, which is characterized in that
Surfactant in the step 3) is Tween 20, one or more in Triton X-100, Bronidox, surface work
Property agent additive amount be 0.01%~0.5%.
4. the kit of sex hormone binding globulin content is measured according to claims 1 or 2 any one of them, it is characterised in that institute
Stating magnetic particle reagent is prepared according to following steps：
1) the carboxyl magnetic bead concentrate after abundant mixing is put into reaction bulb, which is placed in magnetic field to 15~
20min, supernatant is sucked after carboxyl magnetic bead all sedimentation, is added in into reaction bulb and is 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, mixing is for use；
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, mixing state response 18 is kept 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
Required for use magnetic particle reagent is made in 10mg/mL, 2~8 DEG C of preservations.
5. the kit of sex hormone binding globulin content is measured according to claim 4 any one of them, which is characterized in that described
The configuration method of magnetic particle buffer solution be by the Tris of 12.12~15.26mg, the sodium chloride of 5.82~8.58mg, 50~60g's
Methyl cellulose ether is added in 1L purified waters, is stirred well to and is completely dissolved to obtain the final product.
6. the kit according to claim 1 for measuring sex hormone binding globulin content, it is characterised in that：The luminous bottom
Object is fully to dissolve what ALPS was prepared with the luminous substrate buffer solution for being equivalent to 4~10 times of ALPS volumes；The luminous bottom
The configuration method of object buffer solution is to be added to the lucigenin of the sodium chloride of Tris, 5.82g of 12.12g~121.14g, 0.03g
It in 1L purified waters, is stirred well to and is completely dissolved, with salt acid for adjusting pH to 9.5 to obtain the final product.
7. using the method for 2 any one of them kit measurement sex hormone binding globulin content of claims 1 or 2, feature exists
Include the following steps in this method：
1) three test tubes is taken to add 60 μ L calibration objects, 60 μ L quality-control products, 60 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 on magnetic separator is precipitated to 3min, 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) it is primary to repeat step 5)；
7) 200 μ L luminous substrates are added in every test tube, vibrate mixing 3s, luminous intensity is detected with Chemiluminescence Apparatus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711217126.4A CN108226515A (en) | 2017-11-28 | 2017-11-28 | A kind of kit and its test method for measuring sex hormone binding globulin content |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711217126.4A CN108226515A (en) | 2017-11-28 | 2017-11-28 | A kind of kit and its test method for measuring sex hormone binding globulin content |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108226515A true CN108226515A (en) | 2018-06-29 |
Family
ID=62652969
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711217126.4A Pending CN108226515A (en) | 2017-11-28 | 2017-11-28 | A kind of kit and its test method for measuring sex hormone binding globulin content |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108226515A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109856401A (en) * | 2019-03-06 | 2019-06-07 | 苗进超 | A kind of E7 protein detection kit and its detection method and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105223364A (en) * | 2015-10-10 | 2016-01-06 | 广西医科大学 | Serum SHBG albumen is as serum of tuberculosis patients mark and application thereof |
| CN105954266A (en) * | 2016-04-20 | 2016-09-21 | 北京中航赛维生物科技有限公司 | Magnetic particle-based quantitative chemiluminescent assay kit for anti-ribosome P protein antibody IgG, and preparation and detection methods thereof |
| CN106353506A (en) * | 2016-08-12 | 2017-01-25 | 江苏泽成生物技术有限公司 | Kit for detecting osteocalcin content and testing method thereof |
-
2017
- 2017-11-28 CN CN201711217126.4A patent/CN108226515A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105223364A (en) * | 2015-10-10 | 2016-01-06 | 广西医科大学 | Serum SHBG albumen is as serum of tuberculosis patients mark and application thereof |
| CN105954266A (en) * | 2016-04-20 | 2016-09-21 | 北京中航赛维生物科技有限公司 | Magnetic particle-based quantitative chemiluminescent assay kit for anti-ribosome P protein antibody IgG, and preparation and detection methods thereof |
| CN106353506A (en) * | 2016-08-12 | 2017-01-25 | 江苏泽成生物技术有限公司 | Kit for detecting osteocalcin content and testing method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109856401A (en) * | 2019-03-06 | 2019-06-07 | 苗进超 | A kind of E7 protein detection kit and its detection method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108051603A (en) | A kind of kit and its test method for measuring anti-Miao Le Shi pipe hormone-contents | |
| CN106353506A (en) | Kit for detecting osteocalcin content and testing method thereof | |
| CN106290911A (en) | A kind of retinol binding protein measures test kit and method of testing thereof | |
| CN108205059A (en) | A kind of kit and its test method for measuring calcitonin content | |
| CN102998467B (en) | β human chorionic gonadotrophin magnetic microparticle chemiluminescence immune quantitative detection reagent box and preparation method thereof | |
| CN108169486A (en) | A kind of kit and its test method for measuring squamous cell carcinoma-related antigen content | |
| CN106018388B (en) | A kind of pepsinogen Cgene or II assay kit and preparation method thereof | |
| CN107966432A (en) | A kind of Soluble growth that measures stimulates the kit and its test method of 2 protein content of expressing gene | |
| CN102147409B (en) | A kind of method measuring Anti-nucleosome antibodies IgG and reagent device | |
| CN106226526A (en) | A kind of Zinc transporter 8 antibody chemical luminescence immunity detection reagent and preparation method thereof | |
| JPH01227963A (en) | Method and kit for immunoassay | |
| CN102901812A (en) | Magnetic particle chemiluminescence immunoassay kit and assay method for human thyroglobulin antibodies (TGAb) | |
| CN107543932A (en) | The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of calcitonin | |
| CN105092834A (en) | Carbohydrate antigen 19-9 (CA 19-9) quantitative assay kit, preparation method and detection method thereof | |
| CN108931652A (en) | A kind of kit with Magnetism particulate immuno chemistry luminescence method detection myoglobin content | |
| CN110208549A (en) | A kind of Soluble growth stimulation 2 albumen sST2 luminescence reagent box of expressing gene | |
| CN106918708A (en) | A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin | |
| CN106443018A (en) | Myoglobin monoclonal abzyme marking compound and preparation method thereof and detection test kit | |
| CN106248970A (en) | A kind of test kit measuring type i collagen C-terminal peptide content and method of testing thereof | |
| CN111175494A (en) | Thyroglobulin antibody detection kit and use method thereof | |
| CN108226464A (en) | A kind of kit and its test method for measuring thyroglobulin content | |
| CN105445256A (en) | Luteinizing hormone (LH) quantitative determination kit, and making method and detection method thereof | |
| JPH09500215A (en) | Method for measuring analytes and its use for measuring anti-TSH receptor autoantibodies in patient serum | |
| CN113125696A (en) | Estradiol homogeneous phase chemiluminescence detection kit and application thereof | |
| CN106645689A (en) | Thyroid-stimulating hormone receptor antibody chemiluminescent immunoassay kit and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180629 |