CN107677840A - A kind of parathyroid hormone detection kit and its application method - Google Patents
A kind of parathyroid hormone detection kit and its application method Download PDFInfo
- Publication number
- CN107677840A CN107677840A CN201710915772.1A CN201710915772A CN107677840A CN 107677840 A CN107677840 A CN 107677840A CN 201710915772 A CN201710915772 A CN 201710915772A CN 107677840 A CN107677840 A CN 107677840A
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- China
- Prior art keywords
- reagent
- parathyroid hormone
- detection kit
- parathyroid
- polyclonal antibody
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- 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.)
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
-
- 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
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
-
- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/78—Thyroid gland hormones, e.g. T3, T4, TBH, TBG or their receptors
Abstract
The invention discloses a kind of parathyroid hormone detection kit, including reagent R1：MOPSO buffer solutions, NaCl, NaN3, Arabic gum, Tween 80, PEG 2000, solvent is purified water；Reagent R2：MOPSO buffer solutions, NaCl, NaN3, BSA, glycerine, the coated parathyroid gland polyclonal antibody of latex, solvent is purified water；The raw material of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland polyclonal antibody and polyethylene p-chloromethyl styrene copolymer.The invention also discloses a kind of application method of parathyroid hormone detection kit.Advantages of the present invention is：(1) kit of the present invention has accurate, quick compared with high detection sensitivity, simple to operate, quantitative detection；(2) antigen antibody complex that the present invention is formed, good stability, high specificity；(3) it is used for automatic clinical chemistry analyzer, cost is low, pollution-free.
Description
Technical field
The present invention relates to technical field of medical examination, more particularly to a kind of parathyroid hormone detection kit and its use
Method.
Background technology
Parathyroid hormone (parathyroid hormone), it is the alkaline single chain polypeptide of chief cell secretion
Parahormone, abbreviation PTH.Parathyroid hormone is made up of 84 amino acid, and major function is calcium and phosphorus in regulation vertebrate body
Metabolism, promote calcium level to raise, serum phosphorus levels decline.
Synthesize PTH the first precursor, referred to as Pre Pro PTH first in chief cell, contain
115 amino acid, this later precursor crack as by the second precursor first shape containing 90 amino acid in the cell
Glandular hormone is former, and the latter and then in the cell cracking turn into the polypeptide containing 84 amino acid, i.e. PTH.PTH's is dense in human normal plasma
Degree is about 1 nanograms/milliliter.
At present, the method for detecting parathyroid hormone is mainly enzyme linked immunosorbent assay and radio immunoassay.Wherein,
Enzyme linked immunosorbent assay is cumbersome, and quantitative Detection results are bad, and it is larger to be as a result affected by human factors result；Radio-immunity point
Analysis rule has radial pattern pollution, and required instrument is costly.
Therefore, be badly in need of at present it is a kind of it is simple to operate, quantitatively detect the inspection of parathyroid hormone accurate, pollution-free, that cost is low
Test agent box.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of detection simple to operate, quantitative is accurate, nothing
Pollution, cost low parathyroid hormone detection kit and its application method.
The present invention is achieved by the following technical solutions：A kind of parathyroid hormone detection kit, including it is only each other
Vertical reagent R1 and reagent R2 biliquid components, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water；
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland are polyclonal
Antibody and polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
In content be 9%-11%.
One of preferred embodiment as the present invention, including reagent R1 independent of each other and reagent R2 biliquid components, including
Composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water；
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland are polyclonal
Antibody and polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
In content be 10%.
One of preferred embodiment as the present invention, including reagent R1 independent of each other and reagent R2 biliquid components, including
Composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water；
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland are polyclonal
Antibody and polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
In content be 10.5%.
One of preferred embodiment as the present invention, the pH of the MOPSO buffer solutions of the reagent R1 is 6.5.
One of preferred embodiment as the present invention, the pH of the MOPSO buffer solutions of the reagent R2 is 6.5.
A kind of application method of parathyroid hormone detection kit, comprises the following steps：
(1) testing sample is mixed with reagent R1,37 DEG C of incubation 5min；
(2) reacted absorbance A 1 is determined with automatic clinical chemistry analyzer；
(3) mixed again with reagent R2,37 DEG C of reaction 5min；
(4) reacted absorbance A 2 is determined with automatic clinical chemistry analyzer；
(5) concentration of parathyroid hormone in sample is calculated according to absorbance change value.
One of preferred embodiment as the present invention, the reagent R1 of the step (1) and the reagent R2 of step (3) volume ratio
For 3:1.
One of preferred embodiment as the present invention, testing sample and the reagent R1 and reagent R2 compositions of the step (1)
Volume ratio between reagent solution is 1:50.
One of preferred embodiment as the present invention, in the step (2) and step (4), extinction is determined at wavelength 600nm
Spend A1 and A2.
One of preferred embodiment as the present invention, in the step (5), absorbance change value is Δ A, i.e., after A2 subtracts A1
Value.
The present invention compared with prior art the advantages of be：
(1) kit of the present invention has higher detection sensitivity, and simple to operate, quantitative detection is accurate, quick, from detection
Only needed 10 minutes to result is gone out；
(2) antigen antibody complex that the present invention is formed, good stability, there is certain absorbance at a particular wavelength,
High specificity；
(3) present invention can be directly used on automatic clinical chemistry analyzer, coordinate without large-scale instrument and equipment, and cost is low, and
"dead" pollution, it can carry out and promote on a large scale.
Embodiment
Embodiments of the invention are elaborated below, the present embodiment is carried out lower premised on technical solution of the present invention
Implement, give detailed embodiment and specific operating process, but protection scope of the present invention is not limited to following implementation
Example.
Embodiment 1
A kind of parathyroid hormone detection kit of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Body component, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water；
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland are polyclonal
Antibody and polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
In content be 9%.
Embodiment 2
A kind of parathyroid hormone detection kit of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Body component, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water；
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland are polyclonal
Antibody and polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
In content be 11%.
Embodiment 3
A kind of parathyroid hormone detection kit of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Body component, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water；
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland are polyclonal
Antibody and polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
In content be 10%.
Embodiment 4
A kind of parathyroid hormone detection kit of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Body component, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water；
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland are polyclonal
Antibody and polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
In content be 10.5%.
Embodiment 5
The preparation method of parathyroid hormone detection kit in a kind of above-described embodiment of the present embodiment, including following step
Suddenly：
(1) reagent preparation R1：
1. according to the constituent content of reagent R1 in above-described embodiment, MOPSO is dissolved in purified water, stirred, is adjusted
PH is configured to R1 buffer solutions to institute's indicating value is formulated；
2. according to the constituent content of reagent R1 in above-described embodiment, by NaCl, NaN3, Arabic gum, Tween-80, PEG-
2000 are dissolved in R1 buffer solutions, stir, and treat that all raw materials fully dissolve, that is, reagent R1 is made；
(2) reagent preparation R2：
1. according to the constituent content of reagent R2 in above-described embodiment, MOPSO is dissolved in purified water, stirred, is adjusted
PH is configured to R2 buffer solutions to institute's indicating value is formulated；
2. according to the constituent content of reagent R2 in above-described embodiment, by NaCl, NaN3, BSA, glycerine be dissolved in R2 buffer solutions
In, stir, produce R2 dispersion liquids；
3. prepare the coated parathyroid gland polyclonal antibody of latex：
A. the latex microsphere that particle diameter is 80nm and 120nm is taken to add 50g/L EDAC solution in 100mM MES buffer solutions
Mix after being incubated mixing 1h in 37 DEG C of environment, supernatant is removed in centrifugation；Add 50g/L NHS solution to recover to original volume, mix
Even be incubated in 37 DEG C of environment mixes 1h, and centrifugation removes supernatant, must mix microballoon emulsion；
B. polyethylene p-chloromethyl styrene copolymer is added in microballoon emulsion is mixed, is copolymerized 80nm and 120nm particle diameters
Latex；Centrifugation, surplus materials is scattered in MES buffer solutions, 3 times repeatedly, most after recovery in MES buffer solutions to original
Volume, parathyroid gland polyclonal antibody is added, 3.5h is reacted at 37 DEG C, centrifuged, sediment is scattered in the PBS of original volume
In buffer solution, 3 times repeatedly, finally sediment is scattered in the NHS buffer solutions of original volume, adds BSA 30g/L；
C.4 DEG C seal 48h up for safekeeping, obtain the coated parathyroid gland polyclonal antibody of finally required latex；
4. dissolve the obtained coated parathyroid gland polyclonal antibody of latex, ultrasonic disperse, final system with R2 dispersion liquids
Obtain reagent R2.
Embodiment 6
The application method of parathyroid hormone detection kit in a kind of above-described embodiment of the present embodiment, including following step
Suddenly：
(1) 4uL testing samples are mixed with 150uL reagents R1,37 DEG C of incubation 5min；
(2) reacted absorbance A 1 is determined at wavelength 600nm with automatic clinical chemistry analyzer；
(3) mixed again with 50uL reagents R2,37 DEG C of reaction 5min；
(4) reacted absorbance A 2 is determined at wavelength 600nm with automatic clinical chemistry analyzer；
(5) according to absorbance change value Δ A=A2-A1, the concentration of parathyroid hormone in sample is calculated.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should be included in the scope of the protection.
Claims (10)
1. a kind of parathyroid hormone detection kit, it is characterised in that including reagent R1 independent of each other and reagent R2 biliquids
Body component, including composition and corresponding content be：
Reagent R1：
Reagent R2：
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland polyclonal antibody
With polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
Content is 9%-11%.
2. parathyroid hormone detection kit according to claim 1, it is characterised in that including reagent independent of each other
R1 and reagent R2 biliquid components, including composition and corresponding content be：
Reagent R1：
Reagent R2：
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland polyclonal antibody
With polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
Content is 10%.
3. parathyroid hormone detection kit according to claim 1, it is characterised in that including reagent independent of each other
R1 and reagent R2 biliquid components, including composition and corresponding content be：
Reagent R1：
Reagent R2：
The material composition of the coated parathyroid gland polyclonal antibody of latex is：Latex microsphere, parathyroid gland polyclonal antibody
With polyethylene p-chloromethyl styrene copolymer, wherein, polyethylene p-chloromethyl styrene copolymer is in overall reagent R2
Content is 10.5%.
4. according to the parathyroid hormone detection kit described in claim any one of 1-3, it is characterised in that the reagent R1
MOPSO buffer solutions pH be 6.5.
5. according to the parathyroid hormone detection kit described in claim any one of 1-3, it is characterised in that the reagent R2
MOPSO buffer solutions pH be 6.5.
6. a kind of method of parathyroid hormone detection kit using as described in claim any one of 1-3, its feature exist
In comprising the following steps：
(1) testing sample is mixed with reagent R1,37 DEG C of incubation 5min；
(2) reacted absorbance A 1 is determined with automatic clinical chemistry analyzer；
(3) mixed again with reagent R2,37 DEG C of reaction 5min；
(4) reacted absorbance A 2 is determined with automatic clinical chemistry analyzer；
(5) concentration of parathyroid hormone in sample is calculated according to absorbance change value.
7. the application method of parathyroid hormone detection kit according to claim 6, it is characterised in that the step
(1) reagent R1 and the reagent R2 of step (3) volume ratio is 3:1.
8. the application method of parathyroid hormone detection kit according to claim 6, it is characterised in that the step
(1) volume ratio between the reagent solution that testing sample and reagent R1 and reagent R2 is formed is 1:50.
9. the application method of parathyroid hormone detection kit according to claim 6, it is characterised in that the step
(2) and in step (4), absorbance A 1 and A2 are determined at wavelength 600nm.
10. the application method of parathyroid hormone detection kit according to claim 6, it is characterised in that the step
Suddenly in (5), absorbance change value is Δ A, i.e. A2 subtracts the value after A1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710915772.1A CN107677840A (en) | 2017-09-30 | 2017-09-30 | A kind of parathyroid hormone detection kit and its application method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710915772.1A CN107677840A (en) | 2017-09-30 | 2017-09-30 | A kind of parathyroid hormone detection kit and its application method |
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| Publication Number | Publication Date |
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| CN107677840A true CN107677840A (en) | 2018-02-09 |
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| CN201710915772.1A Pending CN107677840A (en) | 2017-09-30 | 2017-09-30 | A kind of parathyroid hormone detection kit and its application method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111596072A (en) * | 2020-06-11 | 2020-08-28 | 安徽大千生物工程有限公司 | Kit for determining PTH based on latex enhanced immunoturbidimetry and preparation and use methods thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103134926A (en) * | 2013-02-27 | 2013-06-05 | 上海交通大学 | Magnetic microsphere carrier and its making method |
| CN105353139A (en) * | 2015-09-28 | 2016-02-24 | 成都博奥新景医学科技有限公司 | Parathyroid hormone quantitative detection kit |
| CN106932588A (en) * | 2015-12-30 | 2017-07-07 | 上海复星长征医学科学有限公司 | Detection α1Kit of-microglobulin and preparation method thereof |
-
2017
- 2017-09-30 CN CN201710915772.1A patent/CN107677840A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103134926A (en) * | 2013-02-27 | 2013-06-05 | 上海交通大学 | Magnetic microsphere carrier and its making method |
| CN105353139A (en) * | 2015-09-28 | 2016-02-24 | 成都博奥新景医学科技有限公司 | Parathyroid hormone quantitative detection kit |
| CN106932588A (en) * | 2015-12-30 | 2017-07-07 | 上海复星长征医学科学有限公司 | Detection α1Kit of-microglobulin and preparation method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111596072A (en) * | 2020-06-11 | 2020-08-28 | 安徽大千生物工程有限公司 | Kit for determining PTH based on latex enhanced immunoturbidimetry and preparation and use methods thereof |
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Application publication date: 20180209 |
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