CN106290820A - A kind of detect test kit of content of glycocholic acid and preparation method thereof in serum - Google Patents
A kind of detect test kit of content of glycocholic acid and preparation method thereof in serum Download PDFInfo
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- CN106290820A CN106290820A CN201610663497.4A CN201610663497A CN106290820A CN 106290820 A CN106290820 A CN 106290820A CN 201610663497 A CN201610663497 A CN 201610663497A CN 106290820 A CN106290820 A CN 106290820A
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- 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
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
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- 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
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/08—Hepato-biliairy disorders other than hepatitis
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Abstract
The invention discloses and a kind of detect the test kit of content of glycocholic acid in serum, including anti-reagent A, anti-reagent B, magnetic particle reagent, calibration object, quality-control product, luminous substrate liquid with concentrate washing liquid；Anti-reagent A is the anti-reagent A of glycocholic acid derivant of alkali phosphatase enzyme mark；Anti-reagent B is the glycocholic acid antibody of FITC labelling, and magnetic particle reagent is the preparation method being coated with and also disclosing this test kit when the carboxyl magnetic particle of FITC antibody confuses.Chemiluminescence is combined by this test kit with immunity magnetic particle, provide a kind of close to homogeneous reaction system, and have employed one-step method reaction pattern, make to detect performance and be greatly improved (sensitivity, elaboration, detection range etc.), response time is greatly shortened, from starting to be loaded onto testing result, the time is less than 35min, hence it is evident that be faster than similar test kit)；Coupling efficiency is high, is firmly combined with, and process stabilizing, while enhancing product performance, greatly reduces product cost.
Description
Technical field
The present invention relates to technological field of biochemistry, be specifically related to a kind of employing Immune competition method, magnetic microparticle chemiluminescence
Method measures the test kit of glycocholic acid (CG) content in human body.
Background technology
Glycocholic acid (Cholyglycine, CG), is the conjunction type cholic acid being combined into by cholic acid and glycine, is bile acid
One of main component.Cholesterol through extremely complex enzymatic reaction, is transformed into primary bile acid in hepatocyte, comprises gallbladder
Acid (CA) and chenodeoxycholic acid (CDCA).When hepatocyte damaged, when liver has disease, just cause glycocholic acid metabolism and circulation disorderly
Disorderly, the liver and gall acid amount in serum is increased by, and relatively ALT, AST, total bilirubin (TBIL), alkali phosphatase (ALP), paddy acyl turn peptide
The conventional liver function index such as enzyme (GGT) and serum albumin (ALB) is the most sensitive.Various hepatic disease are such as: hepatocarcinoma, liver cirrhosis,
The glycocholic acid level of the patient such as acute hepatitis, chronic hepatitis significantly raises, and therefore glycocholic acid can be as diagnosing these diseases
Good index.The detection of glycocholic acid simultaneously has important clinic to the diagnosis of pregnant women's intrahepatic cholestasis (ICP)
Meaning.
The glycocholic acid assay method being currently known has radioimmunology (RIA), enzyme linked immunosorbent assay (ELISA), latex
Strengthen turbidimetry etc..Radioimmunology complex steps, reagent is expensive, need to use supporting instrument and there is radioactivity dirt
Dye.There is detection time length, operation complexity, poor repeatability, be unsuitable for emergency treatment and clinical patient is examined in time in enzyme linked immunosorbent assay
Disconnected needs.Latex enhancing immune turbidimetry is simple to operate, quick, but sensitivity is low, low value poor repeatability.
Summary of the invention
The technical problem to be solved in the present invention is the defect overcoming prior art, it is provided that in a kind of detection serum, glycocholic acid contains
The test kit of amount,
In order to solve above-mentioned technical problem, the invention provides following technical scheme:
The test kit of the present invention includes anti-reagent A, anti-reagent B, magnetic particle reagent, calibration object, quality-control product, luminous substrate liquid
With concentration washing liquid；
Wherein, described anti-reagent A is the glycocholic acid derivant of alkali phosphatase enzyme mark, the anti-reagent A buffer containing BSA；
The glycocholic acid derivant of described alkali phosphatase enzyme mark is by after glycocholic acid derivant and alkali phosphatase coupling by coupling agent
Obtaining, concentration is 0.8ug/mL；Described anti-reagent A buffer is the Tris-HCl solution of 10～100mM, and pH value is 7-8, contains
Concentration is the sodium chloride of PEG-6000,0.7-0.9wt% of 2-6wt%, the sodium azide of 0.05-0.1wt%, the PC-of 0.3wt%
300,0.1-1wt% bovine serum albumin and 20mM EDTA；
Described anti-reagent B is the glycocholic acid antibody of FITC labelling, the anti-reagent B buffer containing BSA；Described FITC labelling
Glycocholic acid antibody is will to be obtained after glycocholic acid antibody and FITC coupling by coupling agent；Described anti-reagent A buffer be 10～
The Tris-HCl solution of 100mM, pH value is the chlorine of PEG-6000,0.7-0.9wt% of ANS, 2-6wt% of 7-8,0.12wt%
Change sodium, the sodium azide of 0.05-0.1wt%, PC-300,0.1-1wt% bovine serum albumin of 0.3wt% and 20mM EDTA；
Described magnetic particle reagent is the carboxyl magnetic particle suspension being coated with FITC antibody；
Described calibration object and quality-control product are made up of humanized's glycocholic acid and buffer, and buffer is concentration 50-200mM
Tris-HCl solution, containing the bovine serum albumin that concentration is 0.1～1wt%, 0.05～the PC-300 of 0.5wt%, 5～50mM
Disodiumedetate；
Described luminous substrate liquid is the Tris-HCl solution containing AMPPD, the pH value of described Tris-HCl solution be 8.0～
10.0, the Tris of KCl and 2.42wt% of NaCl, 0.4wt% of AMPPD, 16wt% containing concentration 20V%；
Further, described coupling agent is selected from carbodiimides, glutaraldehyde, two Carbimide. compounds or dihalide dinitro
One or more in benzene.
Further, described FITC antibody is selected from human serum albumin, bovine serum albumin, cattle transferrins and blood indigo plant egg
One or more in Bai.
Further, described carboxyl magnetic particle is nucleocapsid structure, and diameter range is 100～4500nm, and concentration is 1～5mg/
mL；The surface of described carboxyl magnetic particle is with by one or more modifications of carboxyl, amino, hydroxyl, hydrazide group or chloromethyl
Chemical group；The group bonding that described FITC antibody is carried by carboxyl magnetic particle is on carboxyl magnetic particle surface.
Further, described concentrated cleaning solution is the Tris-HCl solution containing polysorbas20, and pH value is 7.4, containing concentration
The Tris of KCl, 2.42wt% of NaCl, 0.4wt% of 16wt%, with distilled water 15 times dilution during use.
The preparation method of the test kit of the present invention, comprises the steps:
One, the preparation of magnetic particle reagent:
1), by carboxyl magnetic particle 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide and N-hydroxy-succinamide
(NHS) activate under pH4.5～pH5.5, then stand on magnetic field and make carboxyl magnetic particle separate with liquid, discard
Clearly；
2) wash away unnecessaryization activator with the PBS of the 0.01M of pH7.6, be subsequently adding appropriate FITC antibody, make
The concentration of FITC antibody is 0.5ul/mg, concussion reaction under pH7.4～pH7.8；
3) standing on magnetic field after reaction terminates makes carboxyl magnetic particle separate with liquid, and supernatant discarded, with containing 1wt% cattle
The PBS of sero-abluminous 0.01M is closed；
4), after closing terminates, addition confining liquid, to preserve magnetic particle reagent, makes to be coated with the carboxyl magnetic particle of FITC antibody
Ultimate density be 2mg/mL；This magnetic particle suspension is placed in 2-8 DEG C of preservation, in case using；
Two, the preparation of anti-reagent A
The preparation of glycocholic acid derivant
Use SMCC activator to be activated by glycocholic acid, generate maleimide-glycocholic acid derivant；Maleimide
Amine-glycocholic acid derivant contains and can add the alkali phosphatase containing-SH with the maleimide base group of-SH radical reaction,
CG derivant is linked together with alkali phosphatase；
The preparation of anti-reagent A buffer
9gNaCl, 0.5g sodium azide, 3gPC-300,10g bovine serum albumin and 5.84g EDTA are dissolved in the double steaming of 900mL
In water, adjust PH7.0-8.0 with HCl, be settled to 1000mL with distilled water；
Three, the preparation of anti-reagent B
1.2gANS, 9gNaCl, 0.5g sodium azide, 3gPC-300,10g bovine serum albumin and 5.84gEDTA are dissolved in
In 900mL distilled water, adjust pH7.0～8.0 with HCl, be settled to 1000mL with distilled water, it is thus achieved that the buffer containing BSA；
In BSA buffer, under the conditions of 25 DEG C, FITC produces coupling reaction with glycocholic acid antibody, through protein purification instrument
Purification, collects the glycocholic acid antibody being coated with FITC, is coated with the glycocholic acid antibody of FITC through UV spectrophotometer measuring
Concentration, adjust to working concentration be 1.4ug/mL；
Four, calibration object and the preparation of quality-control product
By the bovine serum albumin containing final concentration 0.1-1wt%, the second two of PC-300,5-50mM of 0.05-0.5wt%
The BSA buffer of amine tetraacethyl disodium humanized's glycocholic acid sterling is configured to concentration be 0ug/mL, 1ug/mL, 2.5ug/mL,
A series of calibration objects of 10ug/mL, 20ug/mL, 40ug/mL；
By the bovine serum albumin containing final concentration 0.1-1wt%, the second two of PC-300,5-50mM of 0.05-0.5wt%
Humanized's glycocholic acid sterling is configured to the Quality Control that concentration is 2.5ug/mL, 20ug/mL by the BSA buffer of amine tetraacethyl disodium
Product；
Five, the preparation of luminous substrate liquid
200mL AMPPD, 160gNaCl, 4gKCl, 24.2g trishydroxymethylaminomethane is dissolved in 900mL distilled water,
Adjust PH8.0～10.0 with HCl, be settled to 1000mL with distilled water；
Six, the preparation of concentrated cleaning solution
160NaCl, 4gKCl, 24.2g trishydroxymethylaminomethane, 1mL polysorbas20 are dissolved in 900mL distilled water, use
HCl adjusts to PH7.4, is settled to 1000mL with distilled water, with distilled water 15 times dilution during use.
The know-why of the present invention is as follows:
The glycocholic acid antibody of Fluorescein isothiocyanate (FITC) labelling matches with the glycocholic acid of alkali phosphatase (AP) labelling
Glycocholic acid in antibody and sample, calibration object or quality-control product combines and forms " sandwich " complex.It is subsequently added and is connected with anti-FITC
The magnetic particle of antibody, makes antigenantibody complex be combined in magnetic particle by anti-FITC antibody and the specific binding of FITC
On.Under the effect of externally-applied magnetic field, the complex that immunoreation is formed is separated with other materials unconjugated, clean complex
After, add enzyme-catalyzed chemical luminescence substrate.Substrate by catalytic pyrolysis, forms unstable excited state intermediate under enzyme effect, when
Excited state intermediate returns to send during ground state photon, forms luminescence-producing reaction, can use the luminescence of Chemiluminescence Apparatus detection reaction
Intensity.In detection range, luminous intensity is directly proportional to the content of the glycocholic acid in sample, uses four parameters of improvement
Logistic equation model can calculate glycocholic acid concentration in sample.
It is in place of the main innovation of the present invention:
1, chemiluminescence is combined by this test kit with immunity magnetic particle, it is provided that a kind of close to homogeneous reactant
System, and have employed one-step method reaction pattern so that detection performance is greatly improved (sensitivity, elaboration, detection range etc.), instead
(from starting to be loaded onto testing result, the time is less than 35min, hence it is evident that be faster than similar test kit) it is greatly shortened between Ying Shi；
2, having invented a kind of new FITC antibody and magnetic particle coupling method, the method coupling efficiency is high, is firmly combined with, and
Process stabilizing, while enhancing product performance, greatly reduces product cost.
3, anti-reagent A, anti-reagent B, magnetic particle reagent, calibration object, quality-control product, luminous substrate liquid and concentration in test kit
Washing liquid is all the optimization formula under this reaction system, imitates the phase to the use of this test kit and detection performance provides powerful guarantee.
Accompanying drawing explanation
Accompanying drawing is for providing a further understanding of the present invention, and constitutes a part for description, with the reality of the present invention
Execute example together for explaining the present invention, be not intended that limitation of the present invention.In the accompanying drawings:
Fig. 1 be test kit of the present invention with commercial reagent box A clinical assays result compare figure.
Detailed description of the invention
Hereinafter the preferred embodiments of the present invention are illustrated, 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.
Embodiment
1, sample collection
Correct medical technology is used to collect serum (sample of significant hemolysis or lipidemia cannot be used for measuring), the sample after collection
This is placed in room temperature and may not exceed 8 hours；Sample need to be positioned in the refrigerator of 2-8 DEG C if do not detected in 8 hours；If needing
Within more than 72 hours, preserve or transport, then should be frozen in less than-20 DEG C, it is to avoid multigelation.It is returned to room temperature before using, shakes gently
Dynamic mixing.
2, prepare before experiment
1. take one bottle of concentration washing liquid distilled water and carry out 15 times of dilutions；
2. calorstat or water-bath temperature are adjusted to 37 DEG C, use after temperature stabilization；
3. magnetic particle suspension is fully mixed to being visible by naked eyes precipitation.
3 experimental techniques
1. a certain amount of reaction vessel (flat based tubes) is taken out, numbering.According to requirement of experiment be separately added into 60ul calibration object/
Quality-control product/clinical sample；
2. every hole is separately added into anti-reagent A 60ul, anti-reagent B 60ul；
2. by solution mix homogeneously in reaction vessel, 37 DEG C of incubations 15 minutes；
3. every hole is separately added into the magnetic particle reagent 30ul shaken up；
5. by solution mix homogeneously in reaction vessel, 37 DEG C of incubations 5 minutes；
6. take out reaction vessel, use Magneto separate and washing facility, by the wash liquid 3 times of magnetic particle in reaction vessel；
7. washing liquid, every hole is gone to add luminous substrate liquid 200ul after having washed, concussion；
8. chemiluminescence detector device detection luminous intensity；
9. use four parameter fitting modes, with calibration object concentration value as X-axis, with calibration object luminous intensity values as Y-axis, set up
Calibration curve.Luminous intensity values according to sample to be tested is back-calculated corresponding concentration value.
Test kit of the present invention is identified according to methodology, can reach following index:
Standard curve is linear: R ＞ 0.9900 lowest detectable limit :≤0.07ug/mL.
Table 1 is the testing result of clinical samples, it is known that the lowest detection of test kit of the present invention is limited to≤0.07ug/mL, by
The accurateness of table 2 clinical samples testing result understands the TIANZHU XINGNAO Capsul of test kit of the present invention, and 85%-115% can by table 3
Knowing, coefficient of variation CV≤8% of test kit of the present invention, as shown in Table 4, the coefficient of variation≤15% of test kit of the present invention, by table 5
Understand, the linear dilution of test kit of the present invention: R is more than 0.9900.
Table 1: the testing result of clinical samples
Table 2: the accurateness of clinical samples testing result
Table 3 repeatability quality-control product measured value
The difference between batch of table 4 test kit of the present invention
The linear dilution rate of table 5 test kit of the present invention
120 parts of serum sample measured values are contrasted with commercially available glycocholic acid detection kit A, compares test kit of the present invention with commercially available
The dependency of glycocholic acid test kit testing result., the results are shown in Table 6, accompanying drawing 1 show the test kit of the present invention and commercially available glycocholic acid
The linear equation of detection kit A comparison is y=0.991x+0.061, R=0.998
The test kit of table 6 present invention compares with commercially available glycocholic acid detection kit A
Finally it is noted that the foregoing is only the preferred embodiments of the present invention, it is not limited to the present invention,
Although being described in detail the present invention with reference to previous embodiment, for a person skilled in the art, it still may be used
So that the technical scheme described in foregoing embodiments to be modified, or wherein portion of techniques feature is carried out equivalent.
All within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made, should be included in the present invention's
Within protection domain.
Claims (6)
1. one kind is detected the test kit of content of glycocholic acid in serum, it is characterised in that including: anti-reagent A, anti-reagent B, magnetic particle
Reagent, calibration object, quality-control product, luminous substrate liquid；
Wherein, described anti-reagent A is the glycocholic acid derivant of alkali phosphatase enzyme mark, the anti-reagent A buffer containing BSA；Described
The glycocholic acid derivant of alkali phosphatase enzyme mark is will to be obtained after glycocholic acid derivant and alkali phosphatase coupling by coupling agent,
Concentration is 0.8ug/mL；Described anti-reagent A buffer is the Tris-HCl solution of 10～100mM, and pH value is 7-8, containing concentration
For the sodium chloride of PEG-6000,0.7-0.9wt% of 2-6wt%, the sodium azide of 0.05-0.1wt%, the PC-300 of 0.3wt%,
0.1-1wt% bovine serum albumin and 20mMEDTA；
Described anti-reagent B is the glycocholic acid antibody of FITC labelling, the anti-reagent B buffer containing BSA；The sweet gallbladder of described FITC labelling
Acid antibody is will to be obtained after glycocholic acid antibody and FITC coupling by coupling agent；Described anti-reagent A buffer is 10～100mM
Tris-HCl solution, pH value be PEG-6000,0.7-0.9wt% of ANS, 2-6wt% of 7-8,0.12wt% sodium chloride,
The sodium azide of 0.05-0.1wt%, PC-300,0.1-1wt% bovine serum albumin of 0.3wt% and 20mM EDTA；
Described magnetic particle reagent is the carboxyl magnetic particle suspension being coated with FITC antibody；
Described calibration object and quality-control product are made up of humanized's glycocholic acid and buffer, and buffer is the Tris-of concentration 50-200mM
HCl solution, containing the bovine serum albumin that concentration is 0.1～1wt%, 0.05～the second of the PC-300 of 0.5wt%, 5～50mM
Edetate disodium；
Described luminous substrate liquid is the Tris-HCl solution containing AMPPD, the pH value of described Tris-HCl solution be 8.0～
10.0, the Tris of KCl and 2.42wt% of NaCl, 0.4wt% of AMPPD, 16wt% containing concentration 20V%.
The test kit of content of glycocholic acid in detection serum the most according to claim 1, it is characterised in that described coupling agent selects
One or more in carbodiimides, glutaraldehyde, two Carbimide. compounds or dihalide dinitro benzene.
The test kit of content of glycocholic acid in detection serum the most according to claim 3, it is characterised in that described FITC antibody
One or more in human serum albumin, bovine serum albumin, cattle transferrins and hemocyanin.
The test kit of content of glycocholic acid in detection serum the most according to claim 1, it is characterised in that described carboxyl magnetic is micro-
Grain is nucleocapsid structure, and diameter range is 100～4500nm, and concentration is 1～5mg/mL；The surface of described carboxyl magnetic particle with by
The chemical group that one or more of carboxyl, amino, hydroxyl, hydrazide group or chloromethyl are modified；Described FITC antibody passes through carboxyl
The group bonding that magnetic particle carries is on carboxyl magnetic particle surface.
5. according to the test kit of content of glycocholic acid in the detection serum described in any one of claim 1-4, it is characterised in that also wrap
Including concentrated cleaning solution, described concentrated cleaning solution is the Tris-HCl solution containing polysorbas20, and pH value is 7.4, containing concentration
The Tris of KCl, 2.42wt% of NaCl, 0.4wt% of 16wt%, with distilled water 15 times dilution during use.
6. the preparation method of the test kit of content of glycocholic acid in detection serum as claimed in claim 5, it is characterised in that include
Following steps:
One, the preparation of magnetic particle reagent:
1), by carboxyl magnetic particle 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide and N-hydroxy-succinamide (NHS)
Activate under pH4.5～pH5.5, then stand on magnetic field and make carboxyl magnetic particle separate with liquid, supernatant discarded；
2) wash away unnecessaryization activator with the PBS of the 0.01M of pH7.6, be subsequently adding appropriate FITC antibody, make FITC
The concentration of antibody is 0.5ul/mg, concussion reaction under pH7.4～pH7.8；
3) standing on magnetic field after reaction terminates makes carboxyl magnetic particle separate with liquid, and supernatant discarded, with containing 1wt% Ox blood serum
The PBS of albuminous 0.01M is closed；
4) after closing and terminating, add confining liquid to preserve magnetic particle reagent, make to be coated with the carboxyl magnetic particle of FITC antibody
Final concentration of 2mg/mL；This magnetic particle suspension is placed in 2-8 DEG C of preservation, in case using；
Two, the preparation of anti-reagent A
The preparation of glycocholic acid derivant
Use SMCC activator to be activated by glycocholic acid, generate maleimide-glycocholic acid derivant；Maleimide-sweet
Chlolic acid derivatives contains and can add the alkali phosphatase containing-SH, by CG with the maleimide base group of-SH radical reaction
Derivant links together with alkali phosphatase；
The preparation of anti-reagent A buffer
9gNaCl, 0.5g sodium azide, 3gPC-300,10g bovine serum albumin and 5.84g EDTA are dissolved in 900mL distilled water
In, adjust PH7.0-8.0 with HCl, be settled to 1000mL with distilled water；
Three, the preparation of anti-reagent B
1.2gANS, 9gNaCl, 0.5g sodium azide, 3gPC-300,10g bovine serum albumin and 5.84g EDTA are dissolved in 900mL
In distilled water, adjust pH7.0～8.0 with HCl, be settled to 1000mL with distilled water, it is thus achieved that the buffer containing BSA；
In BSA buffer, under the conditions of 25 DEG C, FITC produces coupling reaction with glycocholic acid antibody, through protein purification instrument purification,
Collect and be coated with the glycocholic acid antibody of FITC, be coated with glycocholic acid antibody dense of FITC through UV spectrophotometer measuring
Degree, adjust to working concentration be 1.4ug/mL；
Four, calibration object and the preparation of quality-control product
With the bovine serum albumin containing final concentration 0.1-1wt%, the ethylenediamine tetraacetic of PC-300,5-50mM of 0.05-0.5wt%
It is 0ug/mL, 1ug/mL, 2.5ug/mL, 10ug/ that humanized's glycocholic acid sterling is configured to concentration by the BSA buffer of acetic acid disodium
A series of calibration objects of mL, 20ug/mL, 40ug/mL；
With the bovine serum albumin containing final concentration 0.1-1wt%, the ethylenediamine tetraacetic of PC-300,5-50mM of 0.05-0.5wt%
Humanized's glycocholic acid sterling is configured to the quality-control product that concentration is 2.5ug/mL, 20ug/mL by the BSA buffer of acetic acid disodium；
Five, the preparation of luminous substrate liquid
200mL AMPPD, 160gNaCl, 4gKCl, 24.2g trishydroxymethylaminomethane is dissolved in 900mL distilled water, uses HCl
Adjust PH8.0～10.0, be settled to 1000mL with distilled water；
Six, the preparation of concentrated cleaning solution
160NaCl, 4gKCl, 24.2g trishydroxymethylaminomethane, 1mL polysorbas20 are dissolved in 900mL distilled water, adjust with HCl
Whole it is settled to 1000mL to PH7.4 with distilled water, with distilled water 15 times dilution during use.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610663497.4A CN106290820A (en) | 2016-08-12 | 2016-08-12 | A kind of detect test kit of content of glycocholic acid and preparation method thereof in serum |
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| CN108196043A (en) * | 2017-11-28 | 2018-06-22 | 泰州泽成生物技术有限公司 | Kit of microdose urine protein content and preparation method thereof in a kind of detection serum |
| CN109212195A (en) * | 2018-10-18 | 2019-01-15 | 郑州安图生物工程股份有限公司 | A kind of kit quickly detecting people AMH using Magnetism particulate immuno chemistry luminescence method |
| CN109444399A (en) * | 2018-10-26 | 2019-03-08 | 重庆中元汇吉生物技术有限公司 | A kind of glycocholic acid detection kit |
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| CN109521005A (en) * | 2018-11-21 | 2019-03-26 | 北京利德曼生化股份有限公司 | A kind of magnetic microparticle chemiluminescence detection kit measuring human body content of glycocholic acid |
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| CN109212195A (en) * | 2018-10-18 | 2019-01-15 | 郑州安图生物工程股份有限公司 | A kind of kit quickly detecting people AMH using Magnetism particulate immuno chemistry luminescence method |
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| CN115420825A (en) * | 2022-08-31 | 2022-12-02 | 北京大学第三医院（北京大学第三临床医学院） | Bile acid detection method and bile acid derivative |
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| CN117030997A (en) * | 2023-08-01 | 2023-11-10 | 美康生物科技股份有限公司 | Homogeneous phase immunoassay method of small molecule compound |
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