CN102809654A - Double-particle compounded C-reactive protein detection kit - Google Patents
Double-particle compounded C-reactive protein detection kit Download PDFInfo
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- CN102809654A CN102809654A CN2012103027755A CN201210302775A CN102809654A CN 102809654 A CN102809654 A CN 102809654A CN 2012103027755 A CN2012103027755 A CN 2012103027755A CN 201210302775 A CN201210302775 A CN 201210302775A CN 102809654 A CN102809654 A CN 102809654A
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Abstract
The invention provides a double-particle compounded C-reactive protein detection kit and relates to the field of in vitro diagnosis of medical immunology. The kit consists of three parts, namely an R1 reagent, an R2 reagent and a calibrator, wherein the R1 reagent is a phosphate buffer system and comprises the following components in mass ratio: 30-80mmol/l of phosphate buffer with the pH value of 7.0-7.6, 60-120mm/l of polyethylene glycol 6000-10000 and 10-20 mmol/L of disodium edta; the R2 reagent is an antibody sensitized grain suspension and comprises compounded CRP (C-Reactive Protein) antibody particles, phosphate buffer with the pH value of 7.0-7.6, disodium edta and the like; and the calibrator is a bovine serum substrate and comprises 0.2-2.2% of sodium azide, 1-10% of tween-20 and 1-3% of bovine serum albumin. The kit provided by the invention adopts an antibody coated with compounded latex particles, is high in sensitivity and can be used for detecting 0.2mg/l of samples and improving the linear value.
Description
Technical field:
The present invention relates to medical immunology in-vitro diagnosis field, be specifically related to a kind of pair of C-reactive protein detection kit that particle is compound.
Background technology:
C reactive protein; Nineteen thirty Tillet and Francis in acute lobar pneumonia patients serum, find can be when calcium ion exists and CPS play precipitation reaction and gain the name; It is human important acute phase reactive protein; Acute stage,, concentration can raise thousands of times, and the CRP half life period in the circulation is 19 hours.Human CRP is produced by liver, and by the ring-type pentamer that five identical subunits rely on non-covalent bonds to form, this characteristic structure makes it range five to gather element (one group of calbindin with immune defense characteristic) family.There is CRP equally in lower animal, as in king crab, freshwater mussel etc., also finding, but might not play acute phase reactive protein.The CRP characteristic reaction is under the condition that calcium ion exists, specificity to combine the phosphocholine group.
It is occur in recent years a kind of comparatively stable, body fluid albumen homogeneous phase immunoturbidimetry detection method accurately that latex particle strengthens turbidimetry.The PETIA method is divided into two kinds substantially.A kind of is the scattering turbidimetry detection method; Another kind is the turbid detection method of transmittance.The ultimate principle of these two kinds of methods is closely similar; It all is surface-crosslinked monoclonal antibody at the polymer latex microballoon; When the crosslinked microballoon that antibody arranged with after antigen combines, can flock together rapidly at short notice, changed the astigmatic performance or the light transmission of reactant liquor.And the reactant liquor astigmatism performance or the change of light transmission (being absorbance) and the concentration of tested antigen have stronger correlativity, can reflect the concentration of tested antigen within the specific limits.The PETIA detection method is the mensuration of in homogeneous reaction system, carrying out antigen, antibody response and result.
Because the content distribution of change of serum C RP differs greatly and whenever is raised to every liter of hundreds of milligram from several milligrams of zero points, its assay method needs higher sensitivity and specificity.At present, immunoturbidimetry is the most general method in common, but immunoturbidimetry highest detection limit can only reach 8mg/l, and linearity can only reach 150mg/l, in clinical practice, exists limitation.
Summary of the invention:
The purpose of this invention is to provide a kind of pair of C-reactive protein detection kit that particle is compound; It adopts compounded latex particle coated antibody, and is highly sensitive, can detect the 0.2mg/l sample; Improved linear value; While has improved the purity of antibody greatly and has tired, and has avoided non-specific responding to greatest extent, and making as a result, accuracy is greatly improved.
In order to solve the existing problem of background technology; The present invention adopts following technical scheme: it is made up of R1 reagent, R2 reagent and calibration object three parts; R1 reagent is phosphate buffer, and its mass ratio consists of: the phosphate buffer 30-80mmol/l of PH=7.0-7.6, Macrogol 6000-1000060-120mm/l and disodium ethylene diamine tetraacetate 10-20mmol/L; R2 reagent is the antibody sensitized particle suspension liquid, and it consists of compound CRP antibody sensitized particle, the phosphate buffer of PH=7.0-7.6 and disodium ethylene diamine tetraacetate etc.; Calibration object is a cow's serum matrix, comprises Sodium azide 0.2-2.2%, Tween-20 1-10%, bovine serum albumin(BSA) 1-3%, the number percent of the long-pending consumption of above-mentioned number percent behaviour serotonin plastid.
2 kinds of compound CRP antibody sensitized particles are characterized as the small grain size latex particle diameter between 40-100nm among the described reagent R2, and the large grain size latex particle diameter is between 150-190nm;
In the described reagent R2 antibody sandwich process, large grain size latex and small grain size latex are with the mixed of 1/3-1/6, and it is 1/10-6/10 that goat anti-human crp's polyclonal antibody and polystyrene latex encapsulate mass ratio.
The preparation method of described reagent R2 is: with the polystyrene latex particles of the polystyrene latex particles of 156nm and the 61nm mixed according to 1: 5, goat anti-human crp's polyclonal antibody and mixed polystyrene latex mix with 50: 100 mass ratio, and damping fluid adopts the phosphate buffer of 0.02M; With 37 ℃ of absorption behind both mixings 8 hours, the antibody on connecting was removed not in dialysis afterwards, adds the glycocoll of confining liquid 0.1% skimmed milk power, 0.2mm; Sealed 2 hours; The centrifugal supernatant that goes is with PH7.4 phosphate buffer 45mmol/l, disodium ethylene diamine tetraacetate 10.2mmol/l; 0.05% skimmed milk power, 0.9%NaN3 is diluted to 0.18%.
Described calibration object preparation method is: with treated cow's serum, add BAS 0.1%, NaN3 0.9% obtains the calibration object dilution, is dissolved in the solution of the similar human serum matrix of preparation with recombinant C RP albumen, prepares the standard items of variable concentrations.
The present invention has following beneficial effect: it adopts compounded latex particle coated antibody, and is highly sensitive, can detect the 0.2mg/l sample; Improved linear value; While has improved the purity of antibody greatly and has tired, and has avoided non-specific responding to greatest extent, and making as a result, accuracy is greatly improved.
Description of drawings:
Fig. 1 is the structural representation of embodiment among the present invention.
Embodiment:
This embodiment is taked following technical scheme: it is made up of R1 reagent, R2 reagent and calibration object three parts; R1 reagent is phosphate buffer, and its mass ratio consists of: the phosphate buffer 30-80mmol/l of PH=7.0-7.6, Macrogol 6000-1000060-120mm/l and disodium ethylene diamine tetraacetate 10-20mmol/L; R2 reagent is the antibody sensitized particle suspension liquid, and it consists of compound CRP antibody sensitized particle, the phosphate buffer of PH=7.0-7.6 and disodium ethylene diamine tetraacetate etc.; Calibration object is a cow's serum matrix, comprises Sodium azide 0.2-2.2%, Tween-20 1-10%, bovine serum albumin(BSA) 1-3%, the number percent of the long-pending consumption of above-mentioned number percent behaviour serotonin plastid.
2 kinds of compound CRP antibody sensitized particles are characterized as the small grain size latex particle diameter between 40-100nm among the described reagent R2, and the large grain size latex particle diameter is between 150-190nm;
In the described reagent R2 antibody sandwich process, large grain size latex and small grain size latex are with the mixed of 1/3-1/6, and it is 1/10-6/10 that goat anti-human crp's polyclonal antibody and polystyrene latex encapsulate mass ratio.
The preparation method of described reagent R2 is: with the polystyrene latex particles of the polystyrene latex particles of 156nm and the 61nm mixed according to 1: 5, goat anti-human crp's polyclonal antibody and mixed polystyrene latex mix with 50: 100 mass ratio, and damping fluid adopts the phosphate buffer of 0.02M; With 37 ℃ of absorption behind both mixings 8 hours, the antibody on connecting was removed not in dialysis afterwards, adds the glycocoll of confining liquid 0.1% skimmed milk power, 0.2mm; Sealed 2 hours; The centrifugal supernatant that goes is with PH7.4 phosphate buffer 45mmol/l, disodium ethylene diamine tetraacetate 10.2mmol/l; 0.05% skimmed milk power, 0.9%NaN3 is diluted to 0.18%.
Described calibration object preparation method is: with treated cow's serum, add BAS 0.1%, NaN3 0.9% obtains the calibration object dilution, is dissolved in the solution of the similar human serum matrix of preparation with recombinant C RP albumen, prepares the standard items of variable concentrations.
This embodiment has following beneficial effect: it adopts compounded latex particle coated antibody; Highly sensitive; Can detect the 0.2mg/l sample, improve linear value, the while has improved the purity of antibody greatly and has tired; Avoided non-specific responding to greatest extent, making as a result, accuracy is greatly improved.
Embodiment 1: correlation test; With reference to Fig. 1: the CRP latex enhancement mode reagent that uses this law invention reagent and contrast agents A company; Adopt automatic 7170 automatic clinical chemistry analyzers that 85 parts of human serums are measured by each autoregressive parameter simultaneously, measured value is carried out correlation analysis.According to above-mentioned " the CRP assay method " in parameter measure, X, Y axle are measured value (the content mg/L of CRP).
Result by Fig. 1 finds out that the facies relationship of two kinds of reagent is R
2=0.987, regression equation is y=0.983x+4.096, and it is good that the result shows that this reagent and import reagent are measured patients serum's correlativity, has excellent specificity and accuracy.
Claims (5)
1. two compound C-reactive protein detection kit of particle; It is characterized in that it is made up of R1 reagent, R2 reagent and calibration object three parts; R1 reagent is phosphate buffer, and its mass ratio consists of: the phosphate buffer 30-80mmol/l of PH=7.0-7.6, Macrogol 6000-1000060-120mm/l and disodium ethylene diamine tetraacetate 10-20mmol/L; R2 reagent is the antibody sensitized particle suspension liquid, and it consists of compound CRP antibody sensitized particle, the phosphate buffer of PH=7.0-7.6 and disodium ethylene diamine tetraacetate etc.; Calibration object is a cow's serum matrix, comprises Sodium azide 0.2-2.2%, Tween-20 1-10%, bovine serum albumin(BSA) 1-3%, the number percent of the long-pending consumption of above-mentioned number percent behaviour serotonin plastid.
2. the C-reactive protein detection kit that a kind of pair of particle according to claim 1 is compound; It is characterized in that 2 kinds of compound CRP antibody sensitized particles among the described reagent R2; Be characterized as the small grain size latex particle diameter between 40-100nm, the large grain size latex particle diameter is between 150-190nm.
3. the C-reactive protein detection kit that a kind of pair of particle according to claim 1 is compound; It is characterized in that in the described reagent R2 antibody sandwich process; Large grain size latex and small grain size latex are with the mixed of 1/3-1/6, and it is 1/10-6/10 that goat anti-human crp's polyclonal antibody and polystyrene latex encapsulate mass ratio.
4. the C-reactive protein detection kit that a kind of pair of particle according to claim 1 is compound is characterized in that the preparation method of described reagent R2 is: with the polystyrene latex particles of the polystyrene latex particles of 156nm and the 61nm mixed according to 1: 5, goat anti-human crp's polyclonal antibody and mixed polystyrene latex mix with 50: 100 mass ratio; Damping fluid adopts the phosphate buffer of 0.02M; With 37 ℃ of absorption behind both mixings 8 hours, the antibody on connecting was removed not in dialysis afterwards, adds the glycocoll of confining liquid 0.1% skimmed milk power, 0.2mm; Sealed 2 hours; The centrifugal supernatant that goes is with PH7.4 phosphate buffer 45mmol/l, disodium ethylene diamine tetraacetate 10.2mmol/l; 0.05% skimmed milk power, 0.9%NaN3 is diluted to 0.18%.
5. the C-reactive protein detection kit that a kind of pair of particle according to claim 1 is compound; It is characterized in that described calibration object preparation method is: with treated cow's serum; Add BAS 0.1%; NaN30.9% obtains the calibration object dilution, is dissolved in the solution of the similar human serum matrix of preparation with recombinant C RP albumen, the standard items of preparation variable concentrations.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103134934A (en) * | 2013-02-27 | 2013-06-05 | 宁波美康生物科技股份有限公司 | Kit for simultaneously detecting retinol-binding protein (RBP) in urine sample and serum sample |
| CN104198725A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Anti-cyclic citrullinated peptide (CCP) antibody detection kit |
| CN107656069A (en) * | 2017-09-28 | 2018-02-02 | 宁波普瑞柏生物技术股份有限公司 | Full-range C reactive protein quantitative detecting reagent and method in whole blood |
| CN108508201A (en) * | 2018-03-27 | 2018-09-07 | 北京九强生物技术股份有限公司 | A kind of carcinomebryonic antigen latex enhancing immune is than turbid kit |
| CN109297919A (en) * | 2018-09-22 | 2019-02-01 | 山东博科生物产业有限公司 | A kind of c reactive protein detection kit of antiheparin interference |
| CN109975555A (en) * | 2019-03-11 | 2019-07-05 | 杭州利安生物科技有限公司 | A kind of reagent, method and high thermal stability c reactive protein kit improving c reactive protein kit thermal stability |
| CN110007074A (en) * | 2019-04-18 | 2019-07-12 | 桂林优利特医疗电子有限公司 | For detecting kit, the preparation method and the usage of c reactive protein |
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| CN101769932A (en) * | 2010-01-26 | 2010-07-07 | 宁波美康生物科技有限公司 | Full-range C-reactive protein detection kit |
| CN101907628A (en) * | 2009-06-08 | 2010-12-08 | 武汉中太生物技术有限公司 | Reagent kit for measuring content of C-reactive protein in whole blood and measuring method thereof |
| US20110151435A1 (en) * | 2009-12-17 | 2011-06-23 | Abaxis, Inc. | Novel assays for detecting analytes in samples and kits and compositions related thereto |
| CN102636653A (en) * | 2012-04-19 | 2012-08-15 | 上海蓝怡科技有限公司 | Compounded latex particle-enveloped cystatin C detection kit |
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2012
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101907628A (en) * | 2009-06-08 | 2010-12-08 | 武汉中太生物技术有限公司 | Reagent kit for measuring content of C-reactive protein in whole blood and measuring method thereof |
| US20110151435A1 (en) * | 2009-12-17 | 2011-06-23 | Abaxis, Inc. | Novel assays for detecting analytes in samples and kits and compositions related thereto |
| CN101769932A (en) * | 2010-01-26 | 2010-07-07 | 宁波美康生物科技有限公司 | Full-range C-reactive protein detection kit |
| CN102636653A (en) * | 2012-04-19 | 2012-08-15 | 上海蓝怡科技有限公司 | Compounded latex particle-enveloped cystatin C detection kit |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103134934A (en) * | 2013-02-27 | 2013-06-05 | 宁波美康生物科技股份有限公司 | Kit for simultaneously detecting retinol-binding protein (RBP) in urine sample and serum sample |
| CN104198725A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Anti-cyclic citrullinated peptide (CCP) antibody detection kit |
| CN104198725B (en) * | 2014-08-14 | 2016-04-06 | 上海睿康生物科技有限公司 | Cyclic citrullinated peptid detection kit |
| CN107656069A (en) * | 2017-09-28 | 2018-02-02 | 宁波普瑞柏生物技术股份有限公司 | Full-range C reactive protein quantitative detecting reagent and method in whole blood |
| CN108508201A (en) * | 2018-03-27 | 2018-09-07 | 北京九强生物技术股份有限公司 | A kind of carcinomebryonic antigen latex enhancing immune is than turbid kit |
| CN109297919A (en) * | 2018-09-22 | 2019-02-01 | 山东博科生物产业有限公司 | A kind of c reactive protein detection kit of antiheparin interference |
| CN109975555A (en) * | 2019-03-11 | 2019-07-05 | 杭州利安生物科技有限公司 | A kind of reagent, method and high thermal stability c reactive protein kit improving c reactive protein kit thermal stability |
| CN109975555B (en) * | 2019-03-11 | 2022-03-08 | 杭州利安生物科技有限公司 | Reagent and method for improving thermal stability of C-reactive protein kit and high-thermal stability C-reactive protein kit |
| CN110007074A (en) * | 2019-04-18 | 2019-07-12 | 桂林优利特医疗电子有限公司 | For detecting kit, the preparation method and the usage of c reactive protein |
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