CN109254159A - A kind of serum amyloid A protein immue quantitative detection reagent box - Google Patents
A kind of serum amyloid A protein immue quantitative detection reagent box Download PDFInfo
- Publication number
- CN109254159A CN109254159A CN201811317973.2A CN201811317973A CN109254159A CN 109254159 A CN109254159 A CN 109254159A CN 201811317973 A CN201811317973 A CN 201811317973A CN 109254159 A CN109254159 A CN 109254159A
- Authority
- CN
- China
- Prior art keywords
- protein
- magnetic particle
- quantitative detection
- serum amyloid
- detection reagent
- 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.)
- Withdrawn
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
-
- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Plasma & Fusion (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of serum amyloid A protein immue quantitative detection reagent boxes, including magnetic particle compound reagent, enzyme conjugates solution, Sample dilution, SAA calibration object and luminous substrate；Magnetic particle coupling in the magnetic particle compound reagent has the anti-human SAA monoclonal antibody of mouse；The enzyme conjugates solution is the anti-human SAA antibody-solutions of mouse of horseradish peroxidase-labeled or the anti-human SAA antibody-solutions of mouse of alkali phosphatase enzyme mark.Kit prepared by the present invention can arrange in pairs or groups full-automatic immune detector, easy to operate, as a result more acurrate, and linear wider, sensitivity is higher, and can directly detect whole blood.Compared with turbidimetry, the interference of blood lipid sample is reduced, can more accurately detect human serum, blood plasma, the SAA in whole blood；Carrying automated system is easy to operate, and flux is big；Using magnetic particle platform, using face is wider, and at low cost and audient is wide.
Description
Technical field
The present invention relates to vitro detection technologies, more particularly, to a kind of serum amyloid A protein immue quantitative detection reagent box.
Background technique
Serum amyloid A protein (SAA) belongs to Acute reaction protein, and inflammation or acute stage of infection are fast in 48-72h
Speed increases, and declines in the convalescence of disease.At present in bacterium, virus infection, atherosclerosis, coronary heart disease, acute grafing
Rejection can detect that Serum SA A is increased in the diseases such as tumour.Especially in certain diseases, such as virus infection, transplanting row
Denounce reaction, coronary heart disease etc., the sensibility of SAA is higher than CRP, therefore the quantitative detection of SAA can provide better reference value for clinic.
Serum amyloid A protein quantitative detection includes a variety of methods and platform, mainly includes immunoturbidimetry, colloidal gold infiltration
Filter method, enzyme linked immunosorbent assay and fluorescence immune chromatography method.Immunoturbidimetry is due to instrument requirements height, and in principle
Existing defects cause the testing result of immunoturbidimetry linearly short, testing result inaccuracy, vulnerable to the influence of blood lipid, and not
Whole blood can be detected；Colloidal gold percolation is suitable for examining by bed, and can detecte whole blood, but this method operating process is excessively
Complexity is unfavorable for staff and largely handles；Enzyme linked immunosorbent assay (ELISA) is measured sensitivity with higher, but
It is that operating process is cumbersome, each measured value is required to draw standard curve, and accuracy error is big, and minute is longer, expensive equipment
And it needs to operate by the personnel of certain professional training, therefore do not meet the clinical requirement quickly detected；Fluorescence immunoassay layer
Limitation of the analysis method due to chromatographing platform itself, accuracy problem cannot be resolved always, and the cost is relatively high, at present
Less appearance in the market.
Summary of the invention
It is more acurrate it is an object of the invention to for defect present in the above-mentioned prior art, provide a kind of testing result,
Linear wider, the higher serum amyloid A protein immue quantitative detection reagent box of sensitivity.
To achieve the above object, the present invention can take following technical proposals:
Serum amyloid A protein immue quantitative detection reagent box of the present invention, including magnetic particle compound reagent, enzyme conjugates are molten
Liquid, Sample dilution, SAA calibration object and luminous substrate；Magnetic particle coupling in the magnetic particle compound reagent has mouse anti-
People's SAA monoclonal antibody；The enzyme conjugates solution is the anti-human SAA antibody-solutions of mouse or alkalinity of horseradish peroxidase-labeled
The anti-human SAA antibody-solutions of the mouse of phosphatase enzyme mark.
The partial size of the magnetic particle is 0.5um-20um, and surface modification has ~ COOH, ~ NH2, benzene mesyl or ~ SH base
Group.
The Sample dilution is Tris-HCl, Tris-NaCl, PBS buffer solution or physiological saline.
It include bovine serum albumin(BSA) or Casein protein in the Sample dilution.
The luminous substrate includes A liquid and B liquid, and wherein the main component of A liquid is hydrogen peroxide, and the main component of B liquid is Shandong
Minot or Derivative of Luminol, when use, mix according to 1:1.
The luminous substrate may be AP luminous substrate or horseradish peroxidase luminous substrate, such as Lumi-Phos
530, AMPPD etc..
The present invention designs quantitative detection human serum amyloid A detection kit by magnetic microparticle chemiluminescence method,
Its cardinal principle are as follows: mouse anti-human monoclonal's antibody is coated with by magnetic particle, in conjunction with the serum amyloid in human serum, blood plasma, whole blood
Sample albumin A, then another plant of anti-human monoclonal antibody of mouse being marked with horseradish peroxidase (HRP) or alkaline phosphatase (AP) formed it is double
Antibody sandwich, in the presence of substrate, catalysis substrate shines horseradish peroxidase, is determined by optical detection system
Amount detection.
Kit prepared by the present invention can arrange in pairs or groups full-automatic immune detector, easy to operate, as a result more acurrate, linearly more
Width, sensitivity is higher, and can directly detect whole blood.Compared with turbidimetry, the interference of blood lipid sample is reduced, it can be more
Accurately detect human serum, blood plasma, the SAA in whole blood；Carrying automated system is easy to operate, and flux is big；It is flat using magnetic particle
Platform, using face is wider, and at low cost and audient is wide.
Detailed description of the invention
Fig. 1 is the linear relationship chart of kit of the present invention.
The high level HOOK of Fig. 2 kit of the present invention schemes.
Fig. 3 is kit of the present invention and Siemens's kit clinical data correlation comparison diagram.
Specific embodiment
More detailed explanation is done to the present invention combined with specific embodiments below, in order to the reason of those skilled in the art
Solution.Unless otherwise specified, the reagent used in the present invention and laboratory apparatus are commercial product, and the method used is this field routine
Method.
Embodiment 1 prepares serum amyloid A protein immue quantitative detection reagent box
1, magnetic particle compound reagent is prepared
Magnetic bead buffer solution 30ul is taken, magnetic bead (partial size about 1um) is blown and beaten repeatedly, is placed on Beads enrichment device, after drawing supernatant, addition
Carbodiimide activation liquid reacts at room temperature 30min ~ 1h；Then the anti-human SAA capture antibody of mouse is added to carry out coupling 2h or stay overnight；
After coupling, as supernatant is drawn on Beads enrichment device, ethanolamine solutions is added and carry out closing 30min；Magnetic bead is also placed in point
From on device, after drawing supernatant, envelope is added and protects liquid (main component is Tris-Nacl buffer, BSA and EDTA), mixes, is placed in
Beads enrichment device abandons supernatant, and after repeating 3-5 times, envelope protects liquid and is settled to 3ml.
2, enzyme conjugates solution is prepared
Suitable enzymic-labelled antibody is taken, is added in enzyme dilution by the dilution ratio of 1:2000, enzyme conjugates solution is configured to；
Enzyme dilution used is 0.02MPBS buffer (pH7.4), wherein contain 2%BSA, 0.2%Prolin-300.
3, Sample dilution is prepared
Sample dilution is used as using 0.02MPBS buffer (pH7.4), wherein contain 2%BSA, 0.2%Prolin-300;It uses
When by sample carry out 200 times dilution.
4, SAA calibration object is prepared
Calibration object dilution is used as using 0.02MPBS buffer (pH7.4, wherein contain 2%BSA, 0.2%Prolin-300), it will
Recombinant antigen is diluted in calibration object, aimed concn be respectively 200mg/l, 100mg/l, 50mg/l, 25mg/l, 5mg/l and
0mg/l。
5, luminous substrate is prepared
A liquid: main component is hydrogen peroxide.
B liquid: main component is luminol or Derivative of Luminol.
The detection method of the kit of the present invention of embodiment 2
1,200 times of 20ul Sample Dilution are taken；
2, the magnetic particle compound reagent of 20ul is added in the sample after taking 20ul to dilute, and piping and druming mixes, and adds the enzyme of 100ul
Conjugate solution, 37 DEG C of incubation 10min；
3,0.02M PBS washing lotion is added in the above mixed liquor, cleans 5 times, substrate (A liquid, each 50ul of B liquid) is added and is then pacifying
Scheme full-automatic immunochemiluminescence instrument A2000 to be detected.
The performance detection of the kit of the present invention of embodiment 3
1, the range of linearity:
High level sample (230mg/l) is subjected to gradient dilution, as a result see the table below and Fig. 1:
Conclusion: according to upper table and Fig. 1, the range of linearity of this kit meets 3mg/l-200mg/l.
2, the HOOK risk of kit of the present invention is examined
Antigen high level is subjected to gradient dilution, as a result as shown in following table and Fig. 2:
Conclusion: according to upper table and Fig. 2, this kit can accomplish 3600mg/l and without obvious " hook-shaped ", HOOK inflection point can be extremely
115200mg/l。
3, the correlation test of kit of the present invention
Kit of the present invention is detected into Siemens's reagent clinic definite value sample, clinical data correlation is as shown in figure 3, result table
It is bright, clinical correlation r > 0.99 of kit of the present invention and Siemens or more.
Claims (6)
1. a kind of serum amyloid A protein immue quantitative detection reagent box, it is characterised in that: including magnetic particle compound reagent, enzyme knot
Polymer solution, Sample dilution, SAA calibration object and luminous substrate；Magnetic particle coupling in the magnetic particle compound reagent
There is the anti-human SAA monoclonal antibody of mouse；The enzyme conjugates solution is the anti-human SAA antibody-solutions of mouse of horseradish peroxidase-labeled
Or the anti-human SAA antibody-solutions of mouse of alkali phosphatase enzyme mark.
2. serum amyloid A protein immue quantitative detection reagent box according to claim 1, it is characterised in that: the magnetic particle
Partial size be 0.5um-20um, surface modification has ~ COOH, ~ NH2, benzene mesyl or ~ SH group.
3. serum amyloid A protein immue quantitative detection reagent box according to claim 1, it is characterised in that: the sample is dilute
Releasing liquid is Tris-HCl, Tris-NaCl, PBS buffer solution or physiological saline.
4. serum amyloid A protein immue quantitative detection reagent box according to claim 3, it is characterised in that: the sample is dilute
Release in liquid includes bovine serum albumin(BSA) or Casein protein.
5. serum amyloid A protein immue quantitative detection reagent box according to claim 1, it is characterised in that: the luminous bottom
Object includes A liquid and B liquid, and wherein the main component of A liquid is hydrogen peroxide, and the main component of B liquid is that luminol or luminol are derivative
Object.
6. serum amyloid A protein immue quantitative detection reagent box according to claim 1, it is characterised in that: the luminous bottom
Object is AP luminous substrate or horseradish peroxidase luminous substrate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811317973.2A CN109254159A (en) | 2018-11-07 | 2018-11-07 | A kind of serum amyloid A protein immue quantitative detection reagent box |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811317973.2A CN109254159A (en) | 2018-11-07 | 2018-11-07 | A kind of serum amyloid A protein immue quantitative detection reagent box |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109254159A true CN109254159A (en) | 2019-01-22 |
Family
ID=65043239
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811317973.2A Withdrawn CN109254159A (en) | 2018-11-07 | 2018-11-07 | A kind of serum amyloid A protein immue quantitative detection reagent box |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109254159A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111289759A (en) * | 2020-03-10 | 2020-06-16 | 苏州翊讯生物科技有限公司 | SAA detection kit and SAA quantitative detection method |
| CN113933506A (en) * | 2021-10-29 | 2022-01-14 | 北京利德曼生化股份有限公司 | Magnetic particle chemiluminescence detection kit for determining content of human chitinase 3-like protein 1(CHI3L1) |
| CN114539401A (en) * | 2022-04-26 | 2022-05-27 | 北京科跃中楷生物技术有限公司 | Magnetic particle marking method and detection kit |
| CN117825708A (en) * | 2024-03-04 | 2024-04-05 | 宁波美康盛德生物科技有限公司 | sFlt-1 detection kit |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108169219A (en) * | 2018-01-15 | 2018-06-15 | 浙江艾明德生物科技有限公司 | A kind of kit and preparation method for quantitatively detecting serum amyloid A protein |
-
2018
- 2018-11-07 CN CN201811317973.2A patent/CN109254159A/en not_active Withdrawn
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108169219A (en) * | 2018-01-15 | 2018-06-15 | 浙江艾明德生物科技有限公司 | A kind of kit and preparation method for quantitatively detecting serum amyloid A protein |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111289759A (en) * | 2020-03-10 | 2020-06-16 | 苏州翊讯生物科技有限公司 | SAA detection kit and SAA quantitative detection method |
| CN111289759B (en) * | 2020-03-10 | 2023-10-27 | 苏州翊讯生物科技有限公司 | SAA detection kit and SAA quantitative detection method |
| CN113933506A (en) * | 2021-10-29 | 2022-01-14 | 北京利德曼生化股份有限公司 | Magnetic particle chemiluminescence detection kit for determining content of human chitinase 3-like protein 1(CHI3L1) |
| CN114539401A (en) * | 2022-04-26 | 2022-05-27 | 北京科跃中楷生物技术有限公司 | Magnetic particle marking method and detection kit |
| CN114539401B (en) * | 2022-04-26 | 2022-06-24 | 北京科跃中楷生物技术有限公司 | Magnetic particle marking method and detection kit |
| CN117825708A (en) * | 2024-03-04 | 2024-04-05 | 宁波美康盛德生物科技有限公司 | sFlt-1 detection kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109254159A (en) | A kind of serum amyloid A protein immue quantitative detection reagent box | |
| CN105548565B (en) | A kind of kit and its preparation and application for being used to detect schizotrypanum cruzi antibody | |
| JP7345002B2 (en) | Tumor marker measurement method and measurement reagent | |
| CN106226516B (en) | A kind of hyperbola calibrates quantitative immunochromatographic detection method | |
| CN102159591A (en) | Monoclonal antibody, and immunoassay using same | |
| CN102901810B (en) | Preparation method of latex particles coated with prostate specific antigen-antibody and PSA enhanced turbidimetric immunophelometry kit | |
| CN105651990A (en) | Chemiluminiscence detection kit for 17alpha-hydroxyprogesterone and preparation method and application thereof | |
| CN112014575B (en) | CYFRA21-1 determination kit and preparation method thereof | |
| CN112679605B (en) | Antibodies or antigen binding fragments thereof against novel coronavirus nucleocapsid proteins and uses thereof | |
| CN108398550B (en) | Composition, chip, preparation method of chip and detection device comprising chip | |
| CN108519487A (en) | A kind of quantitative detecting method and detection kit of interleukin-6 | |
| JP2019528438A (en) | Methods and compositions for assaying vitamin D | |
| CN105277690A (en) | Reagent kit and method for full-automatically measuring antiprotease 3 antibody IgG | |
| JP5276924B2 (en) | Antibody to ochratoxin, affinity column using the antibody, and kit for immunological detection of ochratoxin | |
| CN113150133B (en) | Monoclonal antibodies against SARS-CoV-2 or antigen binding fragments thereof | |
| CN117233378B (en) | Fluorescent immunochromatography test strip for quantitative detection of antibody and preparation method thereof | |
| Warrener et al. | Evaluation of a rapid diagnostic test for measles IgM detection; accuracy and the reliability of visual reading using sera from the measles surveillance programme in Brazil, 2015 | |
| CN110596380A (en) | Method for eliminating HOOK effect and immunochromatography detection card thereof | |
| CN109085358A (en) | Serum amyloid A protein assay kit and production method | |
| CN103558384A (en) | Chemiluminescent kit for simultaneously detecting four tumor markers and preparation method of chemiluminescent kit | |
| JP2003107089A (en) | Ultra-high speed and ultrasensitive measuring method | |
| Gao et al. | Highly specific and sensitive immunoassay for the measurement of prostaglandin E2 in biological fluids | |
| CN104749354B (en) | Test kit detecting four kinds of depression marks and preparation method thereof simultaneously | |
| CN112129933A (en) | Reagent, kit and method for resisting biotin interference in immunoassay system | |
| Nguyen et al. | Correlation of ELISA based with random access serologic immunoassays for identifying adaptive immune response to SARS-CoV-2 |
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 | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20190122 |