CN105158473A - Myeloperoxidase measuring kit - Google Patents
Myeloperoxidase measuring kit Download PDFInfo
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- CN105158473A CN105158473A CN201510588749.7A CN201510588749A CN105158473A CN 105158473 A CN105158473 A CN 105158473A CN 201510588749 A CN201510588749 A CN 201510588749A CN 105158473 A CN105158473 A CN 105158473A
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- mpo
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- polyglycol
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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
- G01N33/577—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
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Abstract
The invention discloses a myeloperoxidase measuring kit. The kit comprises reagents R1 and R2 and a calibration solution, wherein the reagent R1 is a buffer solution with the pH value being 7.0-8.0, and the reagent R2 is an anti-human MPO antibody latex reagent; the calibration solution is recombinant protein containing quantitative MPO or natural MPO extracted from human serum. The kit is characterized by also comprising a reagent R3 used for eliminating chyle, and the reagent R3 includes octyl methoxycinnamate, polyoxypropylene polyoxyethylene propanediol ether and polyethylene glycol-sulfuric chrysanthemum glycan magnesium. According to the method, due to the adding of the reagent R3 for resisting chyle, the interference caused by chyle can be greatly lowered.
Description
Technical field
The present invention relates to a kind of myeloperoxidase enzymatic determination kit.
Background technology
Serum myeloperoxidase (myeloperoxidase, MPO) is more representative inflammation label.
The MPO detection method used clinically at present has: radioimmunology (IRA), and this method directly can survey blood plasma, but this method has environmental pollution and has operator and necessarily has radioactive radiation; Chemoluminescence method, this method comparatively radioimmunology is comparatively responsive, and accurately, but cost is relatively costly, needs supporting instrument just can use; Enzyme immunoassay (ELISA), the shortcomings such as there is complex operation, sample needs pre-service, and detection time is long.And latex enhancing immune turbidimetry has easy and simple to handle, fast, the advantages such as detection sensitivity is high, special, quantitatively accurate, can extensively be approved in clinical practice.
Existing latex immunoturbidimetry reagent, because its Cleaning Principle is the turbidity of the immune complex that assaying reaction is formed, is therefore very easily subject to the impact of chyle interference.The method of current this problem of reply, mainly uses special chyle remover to carry out pre-service to sample.And when large-scale mass survey, health examination examination and clinical patient are checked; often can run into the muddy samples such as a large amount of chyles, haemolysis; now by purchasing sample process agent; these samples are processed separately; greatly will increase the workload of operating personnel; improve operation cost, also for the organization arrangement of clinical examination brings a difficult problem.
Therefore, develop a kind of myeloperoxidase enzymatic determination kit can eliminating the latex immunoturbidimetry of chyle interference and seem very necessary.
Chinese patent literature CN102680676A discloses a kind of myeloperoxidase enzymatic determination kit of turbidimetry for Determination, has easy and simple to handle, fast, and the advantages such as detection sensitivity is high, special, quantitatively accurate, but the problem that still there is chyle interference.
Summary of the invention
The object of the invention is to set up a kind of detection method, by adding of anti-chyle reagent R3, chyle is disturbed and greatly reduces.
Technical scheme of the present invention is:
Myeloperoxidase enzymatic determination kit, comprises reagent R1, R2 and calibration solution; Described reagent R1 is the damping fluid of pH value 7.0-8.0, and described reagent R2 is anti-human MPO antibody latex reagent; The natural MPO that described calibration solution is recombinant protein containing quantitative MPO or extracts from human serum, it is characterized in that, described kit also comprises the reagent R3 for eliminating chyle, and described reagent R3 comprises octyl methoxycinnamate, polyoxyethylene polyoxypropylene propylene glycol and polyglycol-sulfuric acid chrysanthemum glycan magnesium.
Optimally, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20-50
Polyoxyethylene polyoxypropylene propylene glycol 20-100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100-200.
Optimally, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20-50
Polyoxyethylene polyoxypropylene propylene glycol 20-100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100-200
Cellulose acetate-butyrate 20-80.
Optimally, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20-50
Polyoxyethylene polyoxypropylene propylene glycol 20-100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100-200
Sodium cellulose glycolate 20-100
Cellulose acetate-butyrate 20-80.
Inventor draws under study for action, and in the mixing disperse system of the octyl methoxycinnamate of special ratios, polyoxyethylene polyoxypropylene propylene glycol and polyglycol-sulfuric acid chrysanthemum glycan magnesium, chyle sedimentation can occur, and does not affect normal latex turbidimetry simultaneously.
Find in practice, add sodium cellulose glycolate or the cellulose acetate-butyrate of proper proportion, the chyle after precipitation can be made more stable.
Embodiment
With specific embodiment, the present invention will be further described below.
Embodiment 1
Adopt latex enhancing immune turbidimetry to detect a kit for MPO content, comprise reagent R1, R2 and calibration object; Described reagent R1: pH value is the damping fluid of 6.5-8.0, described reagent R2 are anti-human MPO antibody latex reagent; The natural MPO that described standard items are recombinant protein containing quantitative MPO or extract from human serum, R1 reagent in described MPO detection kit mainly comprises damping fluid, inorganic salts, accelerator and antiseptic, in described MPO detection kit, R2 reagent mainly comprises stabilizing agent, anti-human list (many) clonal antibody latex particle, damping fluid, stabilizing agent and antiseptic.Wherein latex beads diameter is 60-150nm.Inorganic salts described in reagent R1 are selected from sodium chloride, magnesium chloride, potassium chloride one or more.Damping fluid is: phosphate buffer, carbonate buffer solution, Glycine-NaOH damping fluid.Accelerator is Macrogol 4000, Macrogol 6000, PEG 8000.Antibody selected by reagent R2 is mouse-anti Human megakaryopoietin monoclonal antibody, goat-anti Human megakaryopoietin monoclonal antibody, one or several mixing of the anti-human MPO monoclonal antibody of rabbit.Also can select mouse-anti Human megakaryopoietin polyclonal antibody, goat-anti Human megakaryopoietin polyclonal antibody, rabbit anti-human MPO polyclonal antibody is wherein a kind of.Stabilizing agent in reagent R2 is selected from bovine albumin, glycocoll, gelatin, polysorbas20, Qu Latong one or several.Reagent R2 is anti-human MPO antibody latex reagent, employing be anti-human MPO antibody and the coupling of latex particle phase.Method through centrifugal (filtration) removes unreacted coupling agent and unreacted antibody, disperses to form in the damping fluid containing stabilizing agent and antiseptic.The preparation of the emulsion reagent of described anti-human MPO antibody comprises the following steps.
Step 1: latex is activated in the solution of PH6.5.
Step 2: wash the carbodiimides removed in solution, add the damping fluid of PH7.5-9.0, add MPO antibody, 37 degree of reactions 4 hours.
Step 3: the immobile liquid of the liquid PH7.5-9.0 of step 2 gained is fixed 6 hours.
Step 4: the sealing liquid of the liquid PH7.5-9.0 of step 3 gained is closed 12 hours.
Step 5: non-binding antibody is removed in the liquid scrubbing of step 4 gained, the buffer solution of solution containing stabilizing agent and antiseptic disperses and get final product.
Damping fluid used in step 1 is PBS damping fluid, MES damping fluid, any one damping fluid in carbonate buffer solution, and concentration at 20-100mmol/L, PH between 5.5-6.5.
Latex used in step 1 can be carboxylated polystyrene latex or amidized polystyrene latex.Its aperture is at 60nm-150nm.
Containing one or more in 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide, 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide salt or N-hydroxy thiosuccinimide in step 1.
In step 2, to remove in solution 1 contained unreacted material, employing be that the filtering membrane in high speed freezing centrifuge or ultra micro aperture filters.
In steps of 5, stabilizing agent used is bSA, gelatin, glycocoll, and one or several phases of skimmed milk power mix.
Described kit also comprises the reagent R3 for eliminating chyle, and described reagent R3 comprises octyl methoxycinnamate, polyoxyethylene polyoxypropylene propylene glycol and polyglycol-sulfuric acid chrysanthemum glycan magnesium.
Optimally, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20
Polyoxyethylene polyoxypropylene propylene glycol 20
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100.
Blank determination is carried out to the serum sample that with the addition of different chyle unit concentration, repeats 3 times, calculate equal deviation.Deviation range is considered as noiseless within ± 10%, and deviation range exceedes ± 10% be considered as interference.
Table 1 do not add R3 reagent serum sample result
Table 2 add R3 reagent serum sample result
Embodiment 2
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 50
Polyoxyethylene polyoxypropylene propylene glycol 100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 200.
Embodiment 3
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 30
Polyoxyethylene polyoxypropylene propylene glycol 40
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 120.
Embodiment 4
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 40
Polyoxyethylene polyoxypropylene propylene glycol 90
Polyglycol-sulfuric acid chrysanthemum glycan magnesium.
Embodiment 5
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20
Polyoxyethylene polyoxypropylene propylene glycol 20
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100
Cellulose acetate-butyrate 20.
Embodiment 6
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 50
Polyoxyethylene polyoxypropylene propylene glycol 100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 200
Cellulose acetate-butyrate 80.
Embodiment 7
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 30
Polyoxyethylene polyoxypropylene propylene glycol 40
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 130
Cellulose acetate-butyrate 30.
Embodiment 8
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 40
Polyoxyethylene polyoxypropylene propylene glycol 80
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 170
Cellulose acetate-butyrate 70.
Embodiment 8
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20
Polyoxyethylene polyoxypropylene propylene glycol 20
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100
Sodium cellulose glycolate 20
Cellulose acetate-butyrate 20.
Embodiment 9
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 50
Polyoxyethylene polyoxypropylene propylene glycol 100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 200
Sodium cellulose glycolate 100
Cellulose acetate-butyrate 80.
Embodiment 10
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 25
Polyoxyethylene polyoxypropylene propylene glycol 35
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 110
Sodium cellulose glycolate 25
Cellulose acetate-butyrate 30.
Embodiment 11
As different from Example 1, the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 45
Polyoxyethylene polyoxypropylene propylene glycol 90
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 180
Sodium cellulose glycolate 80
Cellulose acetate-butyrate 60.
Claims (4)
1. myeloperoxidase enzymatic determination kit, comprises reagent R1, R2 and calibration solution; Described reagent R1 is the damping fluid of pH value 7.0-8.0, and described reagent R2 is anti-human MPO antibody latex reagent; The natural MPO that described calibration solution is recombinant protein containing quantitative MPO or extracts from human serum, it is characterized in that, described kit also comprises the reagent R3 for eliminating chyle, and described reagent R3 comprises octyl methoxycinnamate, polyoxyethylene polyoxypropylene propylene glycol and polyglycol-sulfuric acid chrysanthemum glycan magnesium.
2. require described kit according to right 1, it is characterized in that the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20-50
Polyoxyethylene polyoxypropylene propylene glycol 20-100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100-200.
3. require described kit according to right 1, it is characterized in that the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20-50
Polyoxyethylene polyoxypropylene propylene glycol 20-100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100-200
Cellulose acetate-butyrate 20-80.
4. require described kit according to right 1, it is characterized in that the weight portion of described reagent R3 component consists of:
Octyl methoxycinnamate 20-50
Polyoxyethylene polyoxypropylene propylene glycol 20-100
Polyglycol-sulfuric acid chrysanthemum glycan magnesium 100-200
Sodium cellulose glycolate 20-100
Cellulose acetate-butyrate 20-80.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510588749.7A CN105158473A (en) | 2015-09-16 | 2015-09-16 | Myeloperoxidase measuring kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510588749.7A CN105158473A (en) | 2015-09-16 | 2015-09-16 | Myeloperoxidase measuring kit |
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| CN105158473A true CN105158473A (en) | 2015-12-16 |
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| CN201510588749.7A Pending CN105158473A (en) | 2015-09-16 | 2015-09-16 | Myeloperoxidase measuring kit |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105954523A (en) * | 2016-07-14 | 2016-09-21 | 宏葵生物（中国）有限公司 | Cardiac troponin I detection kit |
| CN106053365A (en) * | 2016-05-26 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | Kit for measuring myeloperoxidase and preparation method of kit |
| CN106896227A (en) * | 2017-04-05 | 2017-06-27 | 济南蓄琪生物技术有限公司 | A kind of myeloperoxidase enzyme detection kit |
| CN113358869A (en) * | 2021-06-04 | 2021-09-07 | 山东博科生物产业有限公司 | Stable and sensitive myeloperoxidase detection kit |
-
2015
- 2015-09-16 CN CN201510588749.7A patent/CN105158473A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 唐伟国 主编: "《医学检验诊断试剂的制备与应用》", 31 October 1996 * |
| 朱征 等: "消除高脂血对临床生化测定影响的方法研究", 《西南军医》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106053365A (en) * | 2016-05-26 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | Kit for measuring myeloperoxidase and preparation method of kit |
| CN105954523A (en) * | 2016-07-14 | 2016-09-21 | 宏葵生物（中国）有限公司 | Cardiac troponin I detection kit |
| CN106896227A (en) * | 2017-04-05 | 2017-06-27 | 济南蓄琪生物技术有限公司 | A kind of myeloperoxidase enzyme detection kit |
| CN113358869A (en) * | 2021-06-04 | 2021-09-07 | 山东博科生物产业有限公司 | Stable and sensitive myeloperoxidase detection kit |
| CN113358869B (en) * | 2021-06-04 | 2022-10-14 | 山东博科生物产业有限公司 | Stable and sensitive myeloperoxidase detection kit |
| WO2022252241A1 (en) * | 2021-06-04 | 2022-12-08 | 山东博科生物产业有限公司 | Stable and sensitive myeloperoxidase detection kit |
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