CN106645694A - Fn (fibronectin) detection kit - Google Patents
Fn (fibronectin) detection kit Download PDFInfo
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- CN106645694A CN106645694A CN201610845853.4A CN201610845853A CN106645694A CN 106645694 A CN106645694 A CN 106645694A CN 201610845853 A CN201610845853 A CN 201610845853A CN 106645694 A CN106645694 A CN 106645694A
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- fibronectin
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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
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
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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
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- Urology & Nephrology (AREA)
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- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses an Fn (fibronectin) detection kit which detects Fn with immunological transmission turbidimetry and belongs to the technical field of clinical in-vitro detection. The kit comprises a reagent R. A buffer solution of the reagent R is replaced with a phosphate buffer solution with the pH being 7.8, meanwhile, 10 mmol/L of EDTA-Na2 is added, the stability of the kit is improved, the linear range is wider, and further popularization and use in the market are facilitated.
Description
Technical field
The present invention relates to clinical vitro detection technical field, more particularly to a kind of fibronectin immunoturbidimetry detection
Kit.
Background technology
Fibronectin（Fibronectin, Fn ）The height on to be one group be distributed widely in blood plasma and various kinds of cell surface
Glycoproteins, with the adhesion adjusted between macrophage, neutrophil chemotactic, promotion cell and its between matrix
Etc. physiological action, their generations with various diseases, development have substantial connection.
Fibronectin is adjusted with non-immunity with disulfide bond by two almost identical subunits
The effect of reason element and a kind of macromolecule glycoprotein with multiple biological activities.Plasma fibronectin measurement contributes to cerebral thrombus
With the antidiastole of cerebral hemorrhage, it is extensive that dynamic observation cerebrovascular patients plasma fibronectin content contributes to the detection state of an illness
Multiple and curative effect.Patients with cor pulmonale acute stage plasma fibronectin is decreased obviously, and the phase that deteriorates extremely reduces, and the paracmasis is progressively extensive
It is multiple.Respiratory failure, pulmonary infection and acid-base disorder person plasma fibronectin are substantially reduced.Early-phase hepatocirrhosis, burst
Hepatic failure and decompensated liver cirrhosis plasma fibronectin are then substantially reduced, oxyhepatitis and chronic persistant hepatitis blood plasma
Fibronectin is normal or low.Uremia and serious renal function impaired subjects plasma fibronectin are substantially less than chronic
The still normal chronic nephritis of pyelonephritis, also significantly lower than renal function.Plasma fibronectin reduce it is more obvious, urea nitrogen and
Creatinine increases more notable, poorer.The fibronectin of primary thrombocytopenic purpura patient increases and blood platelet
Not enhancer bleeding also substantially mitigates, and illustrates that Fn plays an important role in hemostasis.
At present, the detection of plasma fibronectin mainly has latex agglutination test, enzyme immunoassay (EIA) and unidirectional immunity
Diffusion method, these method detection times are longer, and experiment condition is wayward, and precision is relatively low, are not suitable for the place of high-volume sample
Reason and Aulomatizeted Detect.The present invention provides a kind of protein-bonded kit of Immunity transmission turbidity detection fibers, optimizes it
Reaction system so as to simple to operate, reproducible, good stability, the full-automatic detection of batch sample can be carried out, so as to be conducive to
In the market further genralrlization is used.
The content of the invention
The present invention is intended to provide a kind of good stability for the protein-bonded kit of detection fibers, the kit is adopted
Immunity transmission turbidity, compared with conventional kit, stability and linear dependence are good, are conducive to reagent clinically to promote
Using.
General principle：
Fibronectin and goat-anti with Immunity transmission turbidity as measuring principle, after sample mixes with reagent, in sample
People's fibronectin antibody specifically binds, and forms insoluble antigen-antibody complex, makes solution produce turbidity,
Detect under 340nm wavelength, the height of the turbidity is directly proportional in the presence of a certain amount of antibody to the content of antigen.By with it is same
The calibration solution of process compares, and calculates the content of fibronectin in unknown sample.
What invention was obtained through the following steps：
A kind of fibronectin detection kit, it is characterised in that it includes reagent R.Reagent composition is as follows：20mmol/L
PH is 7.8 phosphate buffer, 2mg/mL goat-anti people's fibronectin antibody, 2% Macrogol 6000,2% latex
Grain, 10mmol/L EDTA-Na2, 50mmol/L sodium chloride, 10mmol/L Sodium azides.
Described fibronectin detection kit, it is characterised in that reagent R buffer solutions are 25 DEG C, and pH is 7.8 phosphorus
Phthalate buffer.
Described fibronectin detection kit, it is characterised in that the latex particle is polystyrene, particle diameter is
80-160nm。
Described fibronectin detection kit, it is characterised in that the stabilizer is EDTA- Na2（Ethylenediamine tetraacetic
Acetic acid disodium）.
The described protein-bonded detection method of fibronectin detection kit detection fibers is：Using full-automatic raw
Change analyzer to be measured using end-point method, detection dominant wavelength is 340nm.
The kit of the present invention is carried out on the automatic clinical chemistry analyzer with double reagent function, its specifically used method
It is as follows：
Physiological saline, sample or the μ l of calibration object 3 are added, the reagent R of 300 μ l is added, after preincubate 5min absorbance is read
A1, then reading absorbance A 2 after 5min is reacted, and calculate Δ A.
Beneficial effects of the present invention：
1）Using new buffer system, the stability of reagent is improved；
2）Addition EDTA-Na2, reagent stability is enhanced, and impact will not be produced on the degree of accuracy of reagent；
3）The reagent degree of accuracy and good stability, low price is easy to use, is conducive to reagent further genralrlization in the market.
Description of the drawings
The reagent of Fig. 1 embodiments 1 and the correlation curve figure for compareing group reagent；
The reagent of Fig. 2 embodiments 1 imitates phase stability curve figure with group reagent is compareed；
The reagent of Fig. 3 embodiments 1 with compare group reagent contrasting detection result；
The reagent of Fig. 4 embodiments 1 with compare group reagent linear correlation confirmatory experiment testing result；
The reagent of Fig. 5 embodiments 1 and control group reagent stability confirmatory experiment testing result.
Specific embodiment
The present invention is further described with reference to specific embodiment：
Embodiment 1
A kind of conventional fibronectin detection kit, it includes reagent R.
Reagent R is consisted of：
20mmol/L phosphate buffers pH is 7.8
Goat-anti people fibronectin antibody 2mg/mL
Macrogol 6000 2%
Latex particle 2%
EDTA-Na2 10mmol/L
Sodium chloride 50mmol/L
Sodium azide 10mmol/L.
Reagent R described in the present embodiment need to first prepare phosphate buffer when configuring, and after being transferred to proper pH value, then add other things
Matter.When in use, its assay method is using auspicious 800 automatic clinical chemistry analyzer is stepped, using end to kit described in the present embodiment
Point method is measured, and detection dominant wavelength is 340nm, is operated as follows：
Physiological saline, sample or the μ l of calibration object 3 are added, the reagent R of 300 μ l is added, after preincubate 5min absorbance is read
A1, then reading absorbance A 2 after 5min is reacted, and calculate Δ A.
Δ A=A2- A1
Fibronectin Content（mg/L）=（A determines ÷ A standards）× C standards
Determine：Detection sample A standards：Standard items.
Embodiment 2
Accuracy validation is tested：Using the fibronectin detection reagent of embodiment 1 as experimental group, accreditation is obtained on market
A kind of degree of accuracy is good, the fibronectin detection reagent of good stability is detected as a control group, to 20 clinical serums
Sample is detected that testing result is as shown in Figure 3.Obtain the correlation curve of two kinds of reagents（As shown in Figure 1）, as a result table
Bright, the coefficient correlation of two group reagent boxes is 0.9967, illustrates that both correlations are relatively good.Prove that kit of the present invention adds and more
The component for changing does not result in impact on its accuracy, and kit still keeps the preferable degree of accuracy.
Embodiment 3
Linear dependence checking test：High level sample of the Fibronectin Content for 360mg/L is chosen, is carried out with physiological saline
Be serially diluted, prepare 6 variable concentrations sample, concentration be followed successively by 360mg/dL, 288mg/dL, 216mg/dL, 144mg/dL,
72mg/dL、0mg/dL.It is utilized respectively the reagent of embodiment 1 and control group reagent is detected that the sample of each concentration is determined respectively
Three times, average respectively, testing result is as shown in Figure 4.
As shown in figure 4, embodiment 1 is all higher than 0.990 with group reagent testing result coefficient correlation is compareed, and embodiment 1 is tried
Slightly larger than the coefficient correlation of control group reagent testing result, this shows that reagent of the present invention has more to the coefficient correlation of agent testing result
Good linear dependence.
Embodiment 4
Stability confirmatory experiment：The store reagents in 2 DEG C～8 DEG C, the light protected environment of non-corrosiveness gas, detection embodiment 1 and
The stability of control group reagent.Monthly No. 1 determines same pooled serum sample with two group reagents respectively, determines three times and is averaged
Value, detection data is as shown in Figure 5.
Experimental result shows that the reagent of embodiment 1 is stored 15 months in 2 DEG C～8 DEG C, the light protected environment of non-corrosiveness gas
It is stable, and compare and start unstable after group reagent is stored 12 months in 2 DEG C～8 DEG C, the light protected environment of non-corrosiveness gas, say
Bright reagent of the present invention changes buffer solution and addition EDTA-Na2Rear stability strengthens.
In sum, the fibronectin detection kit that the present invention is provided, by the buffer solution of reagent R pH is replaced by
7.8 phosphate buffer, while adding the EDTA-Na of 10mmol/L2, improve the stability of kit, the range of linearity compared with
Good, the degree of accuracy of reagent is also preferable.Therefore, the fibronectin detection kit that the present invention is provided is conducive to entering in the market
One step is promoted the use of.
Claims (5)
1. a kind of fibronectin detection kit, comprising reagent R, it is characterised in that the reagent composition is as follows：20mmol/
L pH are 7.8 phosphate buffer, 2mg/mL goat-anti people's fibronectin antibody, 2% Macrogol 6000,2% latex
Grain, 10mmol/L EDTA-Na2, 50mmol/L sodium chloride, 10mmol/L Sodium azides.
2. fibronectin detection kit according to claim 1, it is characterised in that reagent R buffer solutions are 25 DEG C,
PH is 7.8 phosphate buffer.
3. fibronectin detection kit according to claim 1, it is characterised in that the latex particle is polyphenyl
Propylene, particle diameter is 80-160nm.
4. fibronectin detection kit according to claim 1, it is characterised in that the stabilizer is EDTA-
Na2（Disodium ethylene diamine tetraacetate）.
5. fibronectin detection kit according to claim 1, it is characterised in that analyzed using full-automatic biochemical
Instrument is measured using end-point method, and detection dominant wavelength is 340nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610845853.4A CN106645694A (en) | 2016-09-24 | 2016-09-24 | Fn (fibronectin) detection kit |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610845853.4A CN106645694A (en) | 2016-09-24 | 2016-09-24 | Fn (fibronectin) detection kit |
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| CN106645694A true CN106645694A (en) | 2017-05-10 |
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| CN201610845853.4A Pending CN106645694A (en) | 2016-09-24 | 2016-09-24 | Fn (fibronectin) detection kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108445225A (en) * | 2018-02-27 | 2018-08-24 | 济南百齐生物技术有限公司 | A kind of immunoturbidimetry fibronectin detection reagent that accuracy is high |
| CN109946295A (en) * | 2017-12-20 | 2019-06-28 | 济南腾奔生物技术有限公司 | A kind of microdose urine protein Immunity transmission turbidity detection kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109946295A (en) * | 2017-12-20 | 2019-06-28 | 济南腾奔生物技术有限公司 | A kind of microdose urine protein Immunity transmission turbidity detection kit |
| CN108445225A (en) * | 2018-02-27 | 2018-08-24 | 济南百齐生物技术有限公司 | A kind of immunoturbidimetry fibronectin detection reagent that accuracy is high |
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