CN109633145A - Use the research method of -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum - Google Patents
Use the research method of -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum Download PDFInfo
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- CN109633145A CN109633145A CN201811472904.9A CN201811472904A CN109633145A CN 109633145 A CN109633145 A CN 109633145A CN 201811472904 A CN201811472904 A CN 201811472904A CN 109633145 A CN109633145 A CN 109633145A
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- rheumatoid arthritis
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- albumen
- early stage
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
-
- 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
- G01N33/5436—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals with ligand physically entrapped within the solid phase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/24—Immunology or allergic disorders
Abstract
The present invention provides a kind of research methods using -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum, it include: 3-4 milliliters of blood that (1) extracts early stage rheumatoid arthritis patients using non-anticoagulant venae vasorum, blood plasma is isolated in condensation naturally；(2) expression of wherein FSTL1 albumen is detected using ELISA kit；(3) the expression data for the FSTL1 albumen that will test and the FSTL1 expression range of normal person compare.The present invention passes through research, follistatin sample albumen -1 in serum can be applied in diagnosis early stage rheumatoid arthritis, the index that the concentration of follistatin sample albumen -1 in serum is diagnosed as one, stability is good, and comparison is obvious, rheumatoid arthritis can be diagnosed early, be conducive to control progression of the disease and treatment.
Description
Technical field
The present invention relates to rheumatoid arthritis technical fields, and in particular to a kind of to use follistatin sample albumen -1 in serum
The research method of aided assessment early stage rheumatoid arthritis.
Background technique
Rheumatoid arthritis is to perplex the disease of many people now, and permanent pain is brought to many patients, because
Rheumatoid arthritis is a kind of chronic disease because unknown.Rheumatoid arthritis may to kidney, internal organ, exhale
Desorption system etc. causes damages, it is also possible to cause joint deformity, therefore, rheumatoid arthritis is more early, and discovery is better, convenient for treatment.
Summary of the invention
Follistatin sample egg in serum is used It is an object of the present invention to solve the deficiency of the existing technology and provide a kind of
The research method of white -1 aided assessment early stage rheumatoid arthritis.
The purpose of the present invention is what is be achieved through the following technical solutions:
A kind of research method using -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum, packet
It includes:
(1) 3-4 milliliters of blood that early stage rheumatoid arthritis patients are extracted using non-anticoagulant venae vasorum, naturally condensation separation
Blood plasma out；
(2) expression of wherein FSTL1 albumen is detected using ELISA kit；
(3) the expression data for the FSTL1 albumen that will test and the FSTL1 expression range of normal person compare.
In the above method, the step (2) is specifically included:
(2.1) 96 orifice plates of previous evening coating are detected and are used for ELISA, capturing antibody with PBS buffer solution, to be diluted to work dense
Degree is added in 96 orifice plates according to every 100 μ L of hole, with ambient temperature overnight after the closing of sealing plate film；
(2.2) the detection same day first absorbs the liquid in hole, carries out plate cleaning；The examination of 300 μ L is finally added in every hole
Dilution agent liquid, room temperature close at least one hour, the plate cleaning step before coming again；
(2.3) sample is taken out in advance from -80 DEG C of refrigerators, is dissolved on ice, then gradient dilution kit to specifications
Interior subsidiary people's FSTL1 standard items albumen is coated with before sequentially adding the standard items of 100 μ L and test sample in 15 minutes
In 96 good holes, it is incubated at room temperature 2 hours after being closed with sealing plate film；
(2.4) first the liquid in hole is absorbed, then carries out plate cleaning；The detection antibody that 100 μ L have diluted is added in every hole,
It is incubated at room temperature 2 hours after being closed with sealing plate film；
(2.5) first the liquid in hole is absorbed, then carries out plate cleaning；The strepto- that 100 μ LHRP lotus roots connection is added in every hole is affine
Plain solution is protected from light incubation 20 minutes；
(2.6) first the liquid in hole is absorbed, then carries out plate cleaning；The Chromogenic Substrate Solution of 100 μ L is added in every hole, keeps away
Light is incubated for 20 minutes；The stop bath of 50 μ L is added in every hole, pats 96 orifice plates and is uniformly mixed；Immediately using microplate luminometer in
Absorbance value is read at 450nm, is carried out data analysis, is obtained the expression data of FSTL1 albumen.
In the above method, step (2.2), step (2.4), (2.5), in (2.6), the method that carries out orifice plate cleaning are as follows: every
Hole is added 400 μ L cleaning buffer solutions and cleans 3 times, each all over all being blotted the cleaning buffer solution in hole with aspirator or in paper handkerchief
On pat dry.
Application of the follistatin sample albumen -1 in early stage Diagnosis of Rheumatoid Arthritis in serum, wherein by ovum in serum
Chalone sample albumen -1 is steeped as the biomarker for diagnosing early stage rheumatoid arthritis, in early stage rheumatoid arthritis patients serum
Follistatin sample albumen -1 concentration be higher than normal range (NR).
In conclusion by adopting the above-described technical solution, compared with prior art, the present invention having the advantage that
The present invention can be applied follistatin sample albumen -1 in serum by research in diagnosis early stage rheumatoid arthritis
In, the index that the concentration of follistatin sample albumen -1 in serum is diagnosed as one, stability is good, and compare it is obvious, can be with
Rheumatoid arthritis is diagnosed early, is conducive to control progression of the disease and treatment.
Specific embodiment
A specific embodiment of the invention is further described in detail below.
- 1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum is used the present invention provides a kind of
Research method, because of the index of early stage rheumatoid arthritis and indefinite, it is more likely that rheumatoid factor (RF) be it is negative, still
Diagnosis early is conducive to control progression of the disease and treatment.
Research method of the invention includes:
(1) 3-4 milliliters of blood that early stage rheumatoid arthritis patients are extracted using non-anticoagulant venae vasorum, naturally condensation separation
Blood plasma out；
(2) expression of wherein FSTL1 albumen is detected using ELISA kit；
(3) the expression data for the FSTL1 albumen that will test and the FSTL1 expression range of normal person compare.
In the above method, the step (2) is specifically included:
(2.1) 96 orifice plates of previous evening coating are detected and are used for ELISA, capturing antibody with PBS buffer solution, to be diluted to work dense
Degree is added in 96 orifice plates according to every 100 μ L of hole, with ambient temperature overnight after the closing of sealing plate film；
(2.2) the detection same day first absorbs the liquid in hole, carries out plate cleaning；The examination of 300 μ L is finally added in every hole
Dilution agent liquid, room temperature close at least one hour, the plate cleaning step before coming again；
(2.3) sample is taken out in advance from -80 DEG C of refrigerators, is dissolved on ice, then gradient dilution kit to specifications
Interior subsidiary people's FSTL1 standard items albumen is coated with before sequentially adding the standard items of 100 μ L and test sample in 15 minutes
In 96 good holes, it is incubated at room temperature 2 hours after being closed with sealing plate film；
(2.4) first the liquid in hole is absorbed, then carries out plate cleaning；The detection antibody that 100 μ L have diluted is added in every hole,
It is incubated at room temperature 2 hours after being closed with sealing plate film；
(2.5) first the liquid in hole is absorbed, then carries out plate cleaning；The strepto- that 100 μ LHRP lotus roots connection is added in every hole is affine
Plain solution is protected from light incubation 20 minutes；
(2.6) first the liquid in hole is absorbed, then carries out plate cleaning；The Chromogenic Substrate Solution of 100 μ L is added in every hole, keeps away
Light is incubated for 20 minutes；The stop bath of 50 μ L is added in every hole, pats 96 orifice plates and is uniformly mixed；Immediately using microplate luminometer in
Absorbance value is read at 450nm, is carried out data analysis, is obtained the expression data of FSTL1 albumen.
In above-mentioned steps (2.2), step (2.4), (2.5), (2.6), the method for progress orifice plate cleaning are as follows: every hole is added
400 μ L cleaning buffer solutions clean 3 times, each all over all being blotted with aspirator by the cleaning buffer solution in hole or clapped on paper handkerchief
It is dry.
By the expression data Yu normal expression range of follistatin sample albumen -1 in early stage rheumatoid arthritis patients serum
It compares and learns, the concentration of the follistatin sample albumen -1 in early stage rheumatoid arthritis patients serum is higher than normal range (NR).
Therefore, follistatin sample albumen -1 in serum can be applied in diagnosis early stage rheumatoid arthritis, by serum
The index that the concentration of middle follistatin sample albumen -1 is diagnosed as one, stability is good, and compare it is obvious, can diagnostics classes early
Rheumathritis is conducive to control progression of the disease and treatment.
Embodiment described above is merely to illustrate technical idea and feature of the invention, in the art its object is to make
Technical staff can understand the content of the present invention and implement it accordingly, patent model of the invention only cannot be limited with the present embodiment
It encloses, i.e., it is all according to same changes or modifications made by disclosed spirit, it still falls in the scope of the patents of the invention.
Claims (4)
1. a kind of research method using -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum, feature
It is, comprising:
(1) 3-4 milliliters of blood that early stage rheumatoid arthritis patients are extracted using non-anticoagulant venae vasorum, naturally condensation separation bleeding
Slurry；
(2) expression of wherein FSTL1 albumen is detected using ELISA kit；
(3) the expression data for the FSTL1 albumen that will test and the FSTL1 expression range of normal person compare.
2. according to claim 1 use -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum
Research method, which is characterized in that the step (2) specifically includes:
(2.1) 96 orifice plates of previous evening coating are detected and are used for ELISA, working concentration is diluted to by antibody is captured with PBS buffer solution, presses
It is added in 96 orifice plates according to every 100 μ L of hole, with ambient temperature overnight after the closing of sealing plate film；
(2.2) the detection same day first absorbs the liquid in hole, carries out plate cleaning；The reagent that 300 μ L are finally added in every hole is dilute
Liquid is released, room temperature closes at least one hour, the plate cleaning step before coming again；
(2.3) sample is taken out in advance from -80 DEG C of refrigerators, is dissolved on ice, then gradient dilution kit is included to specifications
People's FSTL1 standard items albumen of band has been coated with before sequentially adding the standard items of 100 μ L and test sample in 15 minutes
In 96 holes, it is incubated at room temperature 2 hours after being closed with sealing plate film；
(2.4) first the liquid in hole is absorbed, then carries out plate cleaning；The detection antibody that 100 μ L have diluted is added in every hole, with envelope
It is incubated at room temperature 2 hours after the closing of plate film；
(2.5) first the liquid in hole is absorbed, then carries out plate cleaning；The Streptavidin that 100 μ LHRP lotus roots connection is added in every hole is molten
Liquid is protected from light incubation 20 minutes；
(2.6) first the liquid in hole is absorbed, then carries out plate cleaning；The Chromogenic Substrate Solution of 100 μ L is added in every hole, is protected from light and incubates
It educates 20 minutes；The stop bath of 50 μ L is added in every hole, pats 96 orifice plates and is uniformly mixed；Immediately using microplate luminometer in 450nm
Place reads absorbance value, carries out data analysis, obtains the expression data of FSTL1 albumen.
3. according to claim 2 use -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum
Research method, which is characterized in that step (2.2), step (2.4), (2.5), in (2.6), the method that carries out orifice plate cleaning are as follows: every
Hole is added 400 μ L cleaning buffer solutions and cleans 3 times, each all over all being blotted the cleaning buffer solution in hole with aspirator or in paper handkerchief
On pat dry.
4. application of the follistatin sample albumen -1 in early stage Diagnosis of Rheumatoid Arthritis in serum, which is characterized in that by serum
Biomarker of the middle follistatin sample albumen -1 as diagnosis early stage rheumatoid arthritis, early stage rheumatoid arthritis patients blood
The concentration of follistatin sample albumen -1 in clear is higher than normal range (NR).
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| CN201811472904.9A CN109633145A (en) | 2018-12-04 | 2018-12-04 | Use the research method of -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum |
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| CN201811472904.9A CN109633145A (en) | 2018-12-04 | 2018-12-04 | Use the research method of -1 aided assessment early stage rheumatoid arthritis of follistatin sample albumen in serum |
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Citations (6)
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| US20110045507A1 (en) * | 2008-01-29 | 2011-02-24 | University Of Pittsburgh - Of The Commonwealth System Of Higher Education | Fstl-1 as a biomaker of inflammation |
| US20130149712A1 (en) * | 2010-08-05 | 2013-06-13 | University Of Pittsburgh - Of The Commonwealth System Of Higher Education | Follistatin-like protein-1 as a biomarker for inflammatory disorders |
| CN104215777A (en) * | 2014-09-18 | 2014-12-17 | 首都医科大学宣武医院 | Method for detecting hemoglobin-combined phosphorylated alpha-synuclein |
| CN104232590A (en) * | 2014-07-10 | 2014-12-24 | 南开大学 | Monoclonal antibody of follistatin-like protein l and application thereof |
| CN105884870A (en) * | 2016-01-25 | 2016-08-24 | 大连大学 | Method for preparing tetramer from O type foot and mouth disease virus polypeptide |
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2018
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Patent Citations (6)
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| US20110045507A1 (en) * | 2008-01-29 | 2011-02-24 | University Of Pittsburgh - Of The Commonwealth System Of Higher Education | Fstl-1 as a biomaker of inflammation |
| US20130149712A1 (en) * | 2010-08-05 | 2013-06-13 | University Of Pittsburgh - Of The Commonwealth System Of Higher Education | Follistatin-like protein-1 as a biomarker for inflammatory disorders |
| CN104232590A (en) * | 2014-07-10 | 2014-12-24 | 南开大学 | Monoclonal antibody of follistatin-like protein l and application thereof |
| CN104215777A (en) * | 2014-09-18 | 2014-12-17 | 首都医科大学宣武医院 | Method for detecting hemoglobin-combined phosphorylated alpha-synuclein |
| CN105884870A (en) * | 2016-01-25 | 2016-08-24 | 大连大学 | Method for preparing tetramer from O type foot and mouth disease virus polypeptide |
| CN105954521A (en) * | 2016-07-08 | 2016-09-21 | 广州华弘生物科技有限公司 | MTB (Mycobacterium Tuberculosis) infection diagnosis kit |
Non-Patent Citations (3)
| Title |
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| 刘茜 等: "干燥综合征患者血清卵泡抑素样蛋白1表达水平及临床意义", 《临床荟萃》 * |
| 张平 等: "卵泡抑素样蛋白1（FSTL1）在类风湿关节炎（RA）中的作用", 《中华医学会第七次全国中青年检验医学学术会议论文汇编》 * |
| 张红 等: "卵泡抑素样蛋白1在类风湿关节炎中的作用", 《国际检验医学杂志》 * |
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