CN107703308A - A kind of s100 β protein detection kits and its application method - Google Patents
A kind of s100 β protein detection kits and its application method Download PDFInfo
- Publication number
- CN107703308A CN107703308A CN201710917761.7A CN201710917761A CN107703308A CN 107703308 A CN107703308 A CN 107703308A CN 201710917761 A CN201710917761 A CN 201710917761A CN 107703308 A CN107703308 A CN 107703308A
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- Prior art keywords
- reagent
- detection kits
- protein detection
- protein
- application method
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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
- 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
- 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
-
- 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
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
Abstract
The invention discloses a kind of s100 β protein detection kits, including reagent R1：Borate buffer, PEG 6000, sucrose, trehalose, bovine serum albumin(BSA), the preservatives of HY 500, its solvent are purified water；Reagent R2：Borate buffer, bovine serum albumin(BSA), S9, latex microsphere, the preservatives of HY 500, S100 β protein antibodies, PVP, its solvent are purified water.The invention also discloses a kind of application method of s100 β protein detection kits.The present invention is that advantage is：(1) there is higher detection sensitivity and accuracy, it is simple to operate, quick；(2) stability is good, can at least be preserved 12 months at 28 DEG C；(3) it is S100 β protein reagent boxes using Latex-enhanced immunoturbidimetric assay, running cost is low, high specificity；(4) it is adapted to full-automatic testing, can carries out and promote on a large scale.
Description
Technical field
The present invention relates to technical field of medical examination, more particularly to a kind of s100 β protein detection kits and its user
Method.
Background technology
S100 β albumen is called nervous centralis differential protein, and s100 β albumen is described as " the C reaction eggs of brain by some scholars
In vain ".The s100 β albumen of certain physiological concentration has neurotrophic effect, and the s100 β albumen of high concentration has neurotoxicity,
The overexpression of S100 β albumen will increase neurological susceptibility of the brain to hypoxic-ischemic, cause the apoptosis of neuron.
The product on the market for S100 β Protein Detections is mainly with fluoroimmunoassay, ELISA at present
The mode of method is detected, and these method costs are high, the degree of accuracy is low, it is also necessary to is equipped with specific instrument and is carried out cooperation detection, grasps
Make comparisons cumbersome.
Therefore, it is badly in need of one kind at present and uses other detection methods, and low cost, high specificity, high sensitivity, standard can be obtained
The new S100 β Protein Detection products that exactness is high, stability is good, with the content of S100 β albumen in quantitative detection serum.
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 cost is low, high specificity, sensitivity
S100 β protein detection kits high, the degree of accuracy is high, stability is good and its application method.
The present invention is achieved by the following technical solutions：A kind of s100 β protein detection kits, including it is independent of each other
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.
A kind of application method of s100 β protein detection kits, comprises the following steps：
(1) testing sample is mixed with reagent R1,37 DEG C of incubation 1min；
(2) reacted absorbance A 1 is determined with automatic clinical chemistry analyzer；
(3) mixed again with reagent R2,37 DEG C of incubation 5min；
(4) reacted absorbance A 2 is determined with automatic clinical chemistry analyzer；
(5) content of the s100 β albumen 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 2:1.
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), in dominant wavelength 380nm, commplementary wave length
Absorbance A 1 and A2 are determined at 405nm.
One of preferred embodiment as the present invention, in the step (5), absorbance change value is Δ A, i.e., after A1 subtracts A2
Value.
The Cleaning Principle of the present invention：S100 β protein polyclone antibodies, S100 β protein polyclones are coated with latex particle
Agglutinating reaction occurs for the S100 β proteantigens in antibody and sample, forms antigen antibody complex and produces turbidity, the turbidity
Height with the S100 β proteantigens concentration in sample into positive correlation, determine its absorbance and calculated according to calibration curve
The content of S100 β proteantigens.
The present invention compared with prior art the advantages of be：
(1) kit of the present invention has higher detection sensitivity and accuracy, simple to operate, quick, from detecting out
As a result at most only 6 minutes are needed；
(2) the S100 β protein antibodies that the S100 β protein antibodies combination latex microspheres in kit reagent R2 of the present invention obtain
Due to the chemical coupling method of use, stability is good, can at least be preserved 12 months at 2-8 DEG C for the preparation of latex particle；
(3) present invention is the S100 β protein reagent boxes using Latex-enhanced immunoturbidimetric assay, and running cost is low, special
It is different in nature strong；
(4) present invention can be used on automatic clinical chemistry analyzer, is adapted to full-automatic testing, can carry out and push away on a large scale
Extensively.
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 s100 β protein detection kits of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water.
Embodiment 2
A kind of s100 β protein detection kits of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water.
Embodiment 3
A kind of s100 β protein detection kits of the present embodiment, including reagent R1 independent of each other and reagent R2 biliquids
Component, including composition and corresponding content be：
Reagent R1：
Its solvent is purified water；
Reagent R2：
Its solvent is purified water.
Embodiment 4
The preparation method of s100 β protein detection kits, comprises the following steps in a kind of above-described embodiment of the present embodiment：
(1) reagent preparation R1：
1. according to the constituent content of reagent R1 in above-described embodiment, bovine serum albumin(BSA) is dissolved in borate buffer, stirred
Mix uniformly, treat fully to dissolve, be configured to R1 dispersion liquids；
2. according to the constituent content of reagent R1 in above-described embodiment, by PEG-6000, sucrose, trehalose, HY-500 anti-corrosions
Agent is dissolved in R1 dispersion liquids, is stirred, and treats 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, bovine serum albumin(BSA) is dissolved in borate buffer, stirred
Mix uniformly, treat fully to dissolve, be configured to dispersion liquid of R2；
2. according to the constituent content of reagent R2 in above-described embodiment, by S9, HY-500 preservative, latex microsphere, PVP in R2
In dispersion liquid, stir, produce R2 dispersion liquids；
3. the marking process of S100 β protein antibodies：
A. the latex particle that particle diameter is 150nm is cleaned 2 times with 0.5mmol/L PBS, each 15000rpm turns
The lower centrifugation 15min of speed, sucks supernatant；Then it is 3% with 0.5mmol/L PBS diluted latex particle to concentration, often
S9 0.5mg are added in milliliter bulk solution, are mixed, room temperature rocks reaction 0.5 hour, and 10min is centrifuged under 15000rpm rotating speeds,
Supernatant is removed, precipitation is suspended in 50mmol/L borate buffer, ultrasonic disperse；
B. 1.5min is centrifuged under 15000rpm rotating speeds, abandons supernatant, precipitation is suspended in 100mmol/L borate buffer
In liquid so that latex particle ultimate density is 5.0%, and ultrasonic disperse, the S100 β for adding 1/3rd volumes while stirring resist
Body, mix, react at room temperature 2h；
C.15000rpm 35min is centrifuged under rotating speed, abandons supernatant, by one times of 100mmol/L's of the antibody being coated with addition
In borate buffer, ultrasonic disperse adds BSA, 4 DEG C of closing 30min, and centrifugation removes supernatant, adds one times of 100mmol/L boron
Acid buffer dilutes, ultrasonic disperse, and 4 DEG C preserve overnight, obtain finally required S100 β protein antibodies latex particles；
4. dissolving obtained S100 β protein antibodies latex particles with R2 dispersion liquids, make latex particle final concentration therein
For 1.0%, ultrasonic disperse, final obtained reagent R2.
Embodiment 5
The application method of s100 β protein detection kits, comprises the following steps in a kind of above-described embodiment of the present embodiment：
(1) 12 μ L testing samples are mixed with 400 μ L reagents R1,37 DEG C of incubation 1min；
(2) reacted absorbance A 1 is determined at dominant wavelength 380nm, commplementary wave length 405nm with automatic clinical chemistry analyzer；
(3) mixed again with 200 μ L reagents R2,37 DEG C of incubation 5min；
(4) reacted absorbance A 2 is determined at dominant wavelength 380nm, commplementary wave length 405nm with automatic clinical chemistry analyzer；
(5) according to absorbance change value, " Δ A=A1-A2 " calculates the content of the s100 β albumen in sample.
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 (6)
1. a kind of s100 β protein detection kits, it is characterised in that including reagent R1 independent of each other and reagent R2 biliquid groups
Point, including composition and corresponding content be：
Reagent R1：
Reagent R2：
A kind of 2. method using s100 β protein detection kits as claimed in claim 1, it is characterised in that including as follows
Step：
(1) testing sample is mixed with reagent R1,37 DEG C of incubation 1min；
(2) reacted absorbance A 1 is determined with automatic clinical chemistry analyzer；
(3) mixed again with reagent R2,37 DEG C of incubation 5min；
(4) reacted absorbance A 2 is determined with automatic clinical chemistry analyzer；
(5) content of the s100 β albumen in sample is calculated according to absorbance change value.
3. the application method of s100 β protein detection kits according to claim 2, it is characterised in that the step (1)
Reagent R1 and step (3) reagent R2 volume ratio be 2:1.
4. the application method of s100 β protein detection kits according to claim 2, it is characterised in that the step (1)
Testing sample and the reagent solution that forms of reagent R1 and reagent R2 between volume ratio be 1:50.
5. the application method of s100 β protein detection kits according to claim 2, it is characterised in that the step (2)
In step (4), absorbance A 1 and A2 are determined at dominant wavelength 380nm, commplementary wave length 405nm.
6. the application method of s100 β protein detection kits according to claim 2, it is characterised in that the step (5)
In, absorbance change value is Δ A, i.e. A1 subtracts the value after A2.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710917761.7A CN107703308A (en) | 2017-09-30 | 2017-09-30 | A kind of s100 β protein detection kits and its application method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710917761.7A CN107703308A (en) | 2017-09-30 | 2017-09-30 | A kind of s100 β protein detection kits and its application method |
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| CN107703308A true CN107703308A (en) | 2018-02-16 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102161716A (en) * | 2010-12-30 | 2011-08-24 | 北京九强生物技术股份有限公司 | Method and reagent for latex sensitization |
| CN103604931A (en) * | 2013-11-15 | 2014-02-26 | 陆上苏 | Human S100 protein detection reagent and preparation method thereof |
| CN106405106A (en) * | 2016-08-31 | 2017-02-15 | 镇江华测金太医学检验所有限公司 | Chemiluminescence enzyme-linked immunosorbent assay kit of central nervous specific protein |
| 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 CN201710917761.7A patent/CN107703308A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102161716A (en) * | 2010-12-30 | 2011-08-24 | 北京九强生物技术股份有限公司 | Method and reagent for latex sensitization |
| CN103604931A (en) * | 2013-11-15 | 2014-02-26 | 陆上苏 | Human S100 protein detection reagent and preparation method thereof |
| 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 |
| CN106405106A (en) * | 2016-08-31 | 2017-02-15 | 镇江华测金太医学检验所有限公司 | Chemiluminescence enzyme-linked immunosorbent assay kit of central nervous specific protein |
Non-Patent Citations (3)
| Title |
|---|
| 令狐颖等: "血清S100β 蛋白测定在老年认知功能障碍疾病诊断中的意义", 《中国老年学杂志》 * |
| 夏昱等: "血清高敏C一反应蛋白S1 00 13蛋白及NSE与急性脑梗死的关系研究", 《中国急救医学》 * |
| 王菁等: "血清S100β 蛋白及超敏C-反应蛋白含量与急性脑梗死患者病情及预后关系的研究", 《现代检验医学杂志》 * |
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Application publication date: 20180216 |