CN107356764A - A kind of apolipoprotein E detection kit and detection method - Google Patents
A kind of apolipoprotein E detection kit and detection method Download PDFInfo
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- CN107356764A CN107356764A CN201710678282.4A CN201710678282A CN107356764A CN 107356764 A CN107356764 A CN 107356764A CN 201710678282 A CN201710678282 A CN 201710678282A CN 107356764 A CN107356764 A CN 107356764A
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- apolipoprotein
- buffer solution
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- nonidet
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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
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
Abstract
The present invention provides a kind of apolipoprotein E detection kit, it includes reagent R1 and reagent R2, included in wherein reagent R1 in Nonidet P40 1 51g/L, reagent R2 and include the 51g/L of Nonidet P40 1, the 16.5g/L of magnesium salts 0.01, the 20g/L of calcium acetate 0.01 and antibody.Kit antibody performance of the invention is good, reproducible, reagent testing result is accurate, disclosure satisfy that requirement.
Description
Technical field
The present invention relates to medical immunology in-vitro diagnosis field, and in particular to a kind of apolipoprotein E detection kit and detection
Method.
Background technology
Turbidimetry is widely used in clinical examination work.It is immunoturbidimetry that application is most common now.
It is all by observing the formation of sediment, aggegation and the generation of haemolysis that the immuno analytical method of early stage is most of
The presence or absence of specific protein and content in testing sample are analyzed in the light scattering caused by aggregate with measure, such as immune to expand
Scattered, immunoelectrophoresis, direct and brief introduction blood clotting, passive hemagglutination, complement fixation test etc., these detection method costs are low, result is easy
In judgement, technically it is easy to grasp, can be widely used for detecting polytype clinical sample.But because above method operation is numerous
It is trivial, time-consuming and sensitivity and poor accuracy and tend to be eliminated.
Immunoturbidimetry overcomes disadvantages mentioned above, and quantitatively accurate automation can be developed with reference to the demand of clinic
Instrument.Therefore, for immunology detection, immunoturbidimetry has immunology antigen, the specificity of antibody binding, has again
Biotrepy feature, during body fluid can be detected in automation biochemical instruments, the micro test substance particularly in blood,
It is a kind of clinical examination practical technique having very much using future.
Immunoturbidimetry (Turbidimetric inhibition immuno assay) is that antigen-antibody combines dynamic survey
Determine method.Its general principle is：When antigen and antibody react and suitable (the general provision antibody of ratio in special dilution system
It is excessive) when, in the presence of the poly- agent of rush of the soluble immune complex of formation in dilution system, separate out, formed micro- from liquid phase
Grain, makes reaction solution turbidity occur.When antibody concentration is fixed, the amount of the immune complex of formation is with the increasing of amount of antigen in sample
Add and increase, the turbidity of reaction solution is consequently increased.Compareed by the turbidity for determining reaction solution with series of standards product, you can meter
Calculate the content of antigen in sample.
The various detecting instruments developed according to the general principle of immunoturbidimetry, developed have been widely used in clinical inspection
The many aspects tested, it is the basic functional principle of Blood coagulation instrument optical method and immunization；It is the load fat in full-automatic biochemical measure
Albumen, haptens and other protein measuring principles；It can also be applied to microorganism detection simultaneously.By accurately to more
Kind material is quantified, and the diagnosis, treatment and prognosis evaluation to many diseases have larger clinical meaning.
The conventional immunoturbidimetry of clinic because its sample dosage is few, can directly on automatic clinical chemistry analyzer batch sample
Analyze, be simple to operate, but the reagent and method established at present all have some shortcoming and defect, are mainly manifested in：
Antibody is easy to cotton-shaped or flaky precipitate occur during preservation, cause antibody performance be deteriorated, repeatability it is bad,
The coefficient of variation (CV) becomes big, directly contributes reagent testing result inaccuracy, and increases filtration step, complex operation, and filters
Antibody may be removed, influences Detection results, patient is affected, it is impossible to meets requirement.
The content of the invention
To solve the above problems, the invention discloses a kind of apolipoprotein E detection kit and detection method, stable reagent
Property is good, homogeneity is good, testing result accuracy is good, easy to operate, easy to utilize.
For achieving the above object, the present invention provides following technical scheme：
We's invention provides a kind of apolipoprotein E detection kit, wherein including reagent R1 and reagent R2, the reagent
Included in R1 in Nonidet P40 1-51g/L, the reagent R2 comprising Nonidet P40 1-51g/L, magnesium salts
0.01-16.5g/L, calcium acetate 0.01-20g/L and antibody 10-1000mg/L.
A kind of apolipoprotein E detection kit, Nonidet P40 is 5-40g/L in the reagent R1, is preferably
30g/L；Nonidet P40 is 4-41g/L, preferably 32g/L in the reagent R2.
A kind of apolipoprotein E detection kit, the magnesium salts are 0.05-10g/L, preferably 5g/L；The magnesium salts is preferred
For the one or more in magnesium sulfate, magnesium chloride or magnesium acetate.
A kind of apolipoprotein E detection kit, the calcium acetate are 0.03-16.5g/L, preferably 9g/L.
Wherein,
Buffer solution, inorganic salts, preservative and aggregation are also included in the reagent R1；Preferably, in the reagent R1 also
Include buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L and aggregation 1-60g/L.
Buffer solution, inorganic salts and preservative are also included in the reagent R2, it is preferable that also comprising buffering in the reagent R2
Liquid 20-100mmol/L, inorganic salts 1-30g/L and preservative 0.5-1g/L.
A kind of apolipoprotein E detection kit, wherein also including calibration object, the calibration object is calibrated using multiple spot, described
Calibration object includes buffer solution 20-100mmol/L, inorganic salts 1-30g/L, preservative 0.1-2g/L, glucan 0.3-100g/L, sea
Algae sugar 1-100g/L, sucrose 1-100g/L, bovine serum albumin(BSA) 1-100g/L and antigen.
A kind of apolipoprotein E detection kit, wherein comprising including buffer solution in reagent R1 and reagent R2, the reagent R1
The poly- second two of 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/L, ethylphenyl
Comprising buffer solution 20-100mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, anti-in alcohol 1-51g/L, the reagent R2
Body 10-1000mg/L, Nonidet P40 1-51g/L, magnesium sulfate 0.01-16.5g/L, calcium acetate 0.01-20g/L；
Preferably,
Buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, poly- second are included in the reagent R1
In the 1-60g/L of glycol 6000, Nonidet P40 5-40g/L, the reagent R2 comprising buffer solution 20-100mmol/L,
Inorganic salts 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 4-41g/L, magnesium sulfate
0.05-10g/L, calcium acetate 0.03-16.5g/L；
It is highly preferred that
Buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, poly- second are included in the reagent R1
Buffer solution 20-100mmol/L, nothing are included in the 1-60g/L of glycol 6000, Nonidet P40 30g/L, the reagent R2
Machine salt 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 32g/L, magnesium sulfate 5g/L,
Calcium acetate 9g/L.
A kind of apolipoprotein E detection kit,
Preferably, the antibody is goat-anti people, rabbit-anti people, horse is anti-human, mouse is anti-human or other animal anti-human antibodies；
Preferably, the buffer solution be acetate buffer, ammonium chloride buffer, phosphate buffer, TRIS buffer solutions,
One or more in borate buffer, glycine buffer, CAPSO, MOPS or Hepes buffer solution；
Preferably, the inorganic salts are one or both of sodium chloride, potassium chloride；
Preferably, the preservative is Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate or ethyl mercury
One or more in sodium thiosulfate；
Preferably, the aggregation is polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or polyethylene glycol
One or more in 8000；
Present aspect additionally provides a kind of detection method using above-mentioned apolipoprotein E detection kit, comprises the following steps：
(1) add reagent R1 into testing sample to mix, testing sample and reagent R1 are (1-9) by volume：300 add
Enter, 37 DEG C of incubations, under certain wavelength, read absorbance A 1；
(2) add reagent R2 into the mixed liquor of step (1) to mix, reagent R2 and reagent R1 is 1 by volume：(1-6)
Add, 37 DEG C of incubations, under certain wavelength, read absorbance A 2；
(3) absorbance △ A, △ A=A2-A1 are drawn；
(4) built-in curve matching model fitting standard curve, root are passed through on automatic clinical chemistry analyzer using calibration object
Calculate the content of apo E in testing sample automatically according to absorbance.
Signified aggregation is the material for promoting antigen-antibody aggegation in the reaction in the present invention.
It is as follows to be related to raw material sources in present invention offer apolipoprotein E detection kit and detection method：
By adopting the above-described technical solution, the beneficial effects of the invention are as follows：
Apolipoprotein E detection kit provided by the invention and detection method, reagent stability is good, and homogeneity is good, antibody
It can stablize and preserve, be not in deposited phenomenon, antibody titer is high, performance is good, and does not have filtration step, easy to operate, into
This is cheap, and testing result is accurate, reproducible, has wider versatility.
Embodiment
In order that those skilled in the art more fully understand the technical scheme in the application, with reference to embodiment to this hair
It is bright to be described further, it is clear that described embodiment is only some embodiments of the present application, rather than whole implementation
Example.Based on the embodiment in the application, what those of ordinary skill in the art were obtained under the premise of creative work is not made
Every other embodiment, it should all belong to the scope of the application protection.
The apolipoprotein E detection kit of embodiment 1
Reagent R1：
| TRIS buffer solutions | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 1g/L |
Reagent R2：
| TRIS buffer solutions | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein E antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Magnesium sulfate | 0.01g/L |
| Calcium acetate | 0.01g/L |
Calibration object：
ApoE antigens with standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L Sodium azides,
0.3g/L glucans, 1g/L trehaloses, 2g/L sucrose, 2g/L bovine serum albumin(BSA)s) dissolving, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, and with standard dilutions be diluted to various concentrations ApoE mark
Quasi- product (ApoE antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then removed with 0.65 μm of membrane filtration
Bacterium, place 2~8 DEG C of preservations.
The apolipoprotein E detection kit of embodiment 2
Reagent R1：
| Phosphate buffer | 40mmol/L |
| Potassium chloride | 7g/L |
| Phenol | 0.7g/L |
| Macrogol 6000 | 30g/L |
| Nonidet P40 | 51g/L |
Reagent R2：
Calibration object：
ApoE antigens with standard dilutions (50mmol/L glycine buffers, 10g/L sodium chloride, 0.5g/L Sodium azides,
20g/L glucans, 25g/L trehaloses, 30g/L sucrose, 20g/L bovine serum albumin(BSA)s) dissolving, with the detection of commercially available contrast agents simultaneously
Adjust to 100mg/L, packing and be stored in -20 DEG C.Using preceding taking-up, and it is diluted to standard dilutions the ApoE of various concentrations
Standard items (ApoE antigen concentrations：0mg/L、10mg/L、30mg/L、50mg/L、70mg/L).Then with 0.65 μm of membrane filtration
It is degerming, place 2~8 DEG C of preservations.
The apolipoprotein E detection kit of embodiment 3
Reagent R1：
| Acetate buffer | 80mmol/L |
| Potassium chloride | 20g/L |
| Sodium azide | 0.8g/L |
| Macrogol 6000 | 45g/L |
| Nonidet P40 | 5g/L |
Reagent R2：
| Acetate buffer | 80mmol/L |
| Potassium chloride | 20g/L |
| Sodium azide | 1g/L |
| Goat-anti human apolipoprotein E antibody | 300mg/L |
| Nonidet P40 | 4g/L |
| Magnesium sulfate | 0.05g/L |
| Calcium acetate | 0.03g/L |
Calibration object：
ApoE antigens with standard dilutions (70mmol/L TRIS buffer solutions, 25g/L sodium chloride, 1g/L Sodium azides,
55g/L glucans, 50g/L trehaloses, 60g/L sucrose, 75g/L bovine serum albumin(BSA)s) dissolving, with the detection of commercially available contrast agents simultaneously
Adjust to 200mg/L, packing and be stored in -20 DEG C.Using preceding taking-up, and it is diluted to standard dilutions the ApoE of various concentrations
Standard items (ApoE antigen concentrations：2mg/L、10mg/L、40mg/L、60mg/L、80mg/L).Then with 0.65 μm of membrane filtration
It is degerming, place 2~8 DEG C of preservations.
The apolipoprotein E detection kit of embodiment 4
Reagent R1：
| MOPS buffer solutions | 150mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Macrogol 6000 | 55g/L |
| Nonidet P40 | 40g/L |
Reagent R2：
| MOPS buffer solutions | 100mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Goat-anti human apolipoprotein E antibody | 600mg/L |
| Nonidet P40 | 41g/L |
| Magnesium sulfate | 10g/L |
| Calcium acetate | 16.5g/L |
Calibration object：
ApoE antigens with standard dilutions (100mmol/L TRIS buffer solutions, 15g/L sodium chloride, 1g/L Sodium azides,
10g/L glucans, 100g/L trehaloses, 95g/L sucrose, 100g/L bovine serum albumin(BSA)s) dissolving, is detected with commercially available contrast agents
And adjust to 160mg/L, packing and be stored in -20 DEG C.Various concentrations are diluted to using preceding taking-up, and with standard dilutions
ApoE standard items (ApoE antigen concentrations：0mg/L、20mg/L、40mg/L、70mg/L、100mg/L).Then with 0.45 μm of filter
Membrane filtration is degerming, places 2~8 DEG C of preservations.
The apolipoprotein E detection kit of embodiment 5
Reagent R1：
| MOPS buffer solutions | 150mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Macrogol 6000 | 60g/L |
| Nonidet P40 | 30g/L |
Reagent R2：
| MOPS buffer solutions | 100mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Goat-anti human apolipoprotein E antibody | 1000mg/L |
| Nonidet P40 | 32g/L |
| Magnesium sulfate | 5g/L |
| Calcium acetate | 9g/L |
Calibration object：
ApoE antigens with standard dilutions (100mmol/L TRIS buffer solutions, 30g/L sodium chloride, 0.5g/L Sodium azides,
90g/L glucans, 100g/L trehaloses, 100g/L sucrose, 100g/L bovine serum albumin(BSA)s) dissolving, examined with commercially available contrast agents
Survey and adjust to 120mg/L, packing and be stored in -20 DEG C.Various concentrations are diluted to using preceding taking-up, and with standard dilutions
ApoE standard items (ApoE antigen concentrations：0mg/L、20mg/L、50mg/L、80mg/L、100mg/L).Then with 0.65 μm of filter
Membrane filtration is degerming, places 2~8 DEG C of preservations.
The apolipoprotein E detection kit of embodiment 6
Reagent R1：
Reagent R2：
| TRIS buffer solutions | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein E antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Magnesium sulfate | 0.01g/L |
Calibration object：
ApoE antigens with standard dilutions (25mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L Sodium azides,
0.3g/L glucans, 1g/L trehaloses, 3g/L sucrose, 3g/L bovine serum albumin(BSA)s) dissolving, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, and with standard dilutions be diluted to various concentrations ApoE mark
Quasi- product (ApoE antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then removed with 0.65 μm of membrane filtration
Bacterium, place 2~8 DEG C of preservations.
The apolipoprotein E detection kit of embodiment 7
Reagent R1：
| TRIS buffer solutions | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 1g/L |
Reagent R2：
| TRIS buffer solutions | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein E antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Calcium acetate | 0.01g/L |
Calibration object：
ApoE antigens with standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L Sodium azides,
0.3g/L glucans, 1g/L trehaloses, 2g/L sucrose, 2g/L bovine serum albumin(BSA)s) dissolving, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, and with standard dilutions be diluted to various concentrations ApoE mark
Quasi- product (ApoE antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then removed with 0.65 μm of membrane filtration
Bacterium, place 2~8 DEG C of preservations.
The apolipoprotein E detection kit of embodiment 8
Reagent R1：
| TRIS buffer solutions | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Polysorbas20 | 1g/L |
Reagent R2：
| TRIS buffer solutions | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein E antibody | 10mg/L |
| Polysorbas20 | 1g/L |
Calibration object：
ApoE antigens with standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L Sodium azides,
0.3g/L glucans, 1g/L trehaloses, 2g/L sucrose, 2g/L bovine serum albumin(BSA)s) dissolving, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, and with standard dilutions be diluted to various concentrations ApoE mark
Quasi- product (ApoE antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then removed with 0.65 μm of membrane filtration
Bacterium, place 2~8 DEG C of preservations.Different embodiment testing results compare, wherein CV values=STDEV (1-7)/average.
1. preparing the sample that a apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 1
7 times, testing result is as shown in table 1.
Table 1：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 1, apo E concentration that embodiment 1 measured 1 month, 3 months, 12 months close to actual value,
And the coefficient of variation (CV values) measured) 2% is respectively less than, illustrate that the kit of the present invention is reproducible, stable performance, measurement is accurate
Really.
2. preparing the sample that a apo E concentration is 1mg/L, detection 7 is repeated to sample with the kit of embodiment 2
Secondary, testing result is as shown in table 2.
Table 2：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 2, apo E concentration that embodiment 2 measured 1 month, 3 months, 12 months close to actual value,
And the coefficient of variation measured is respectively less than 2%, illustrate that the kit of the present invention is reproducible, stable performance, measurement is accurate.
3. preparing the sample that a apo E concentration is 25mg/L, sample is repeated to detect with the kit of embodiment 3
7 times, testing result is as shown in table 3.
Table 3：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 3, apo E concentration that embodiment 3 measured 1 month, 3 months, 12 months close to actual value,
And the coefficient of variation measured is respectively less than 1%, illustrate that the kit of the present invention is reproducible, stable performance, measurement is accurate.
4. preparing the sample that a apo E concentration is 70mg/L, sample is repeated to detect with the kit of embodiment 4
7 times, testing result is as shown in table 4.
Table 4：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 4, apo E concentration that embodiment 4 measured 1 month, 3 months, 12 months close to actual value,
And the coefficient of variation measured is respectively less than 1%, illustrate that the kit of the present invention is reproducible, stable performance, measurement is accurate.
5. preparing the sample that a apo E concentration is 25mg/L, sample is repeated to detect with the kit of embodiment 5
7 times, testing result is as shown in table 5.
Table 5：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 5, embodiment 5 is actual value in the apo E concentration that 1 month, 3 months, 12 months measure, and
And the coefficient of variation measured is respectively less than 0.05%, illustrate that the kit of the present invention is reproducible, stable performance, measurement is accurate.
6. preparing the sample that a apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 6
7 times, testing result is as shown in table 6.
Table 6：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 6, embodiment 6 deviates actual value in the apo E concentration that 1 month, 3 months, 12 months measure,
And the coefficient of variation measured is all higher than 4%, illustrate that the kit repeatability is bad, performance is unstable, measurement is inaccurate.
7. preparing the sample that a apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 7
7 times, testing result is as shown in table 7.
Table 7：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 7, embodiment 7 deviates actual value in the apo E concentration that 1 month, 3 months, 12 months measure,
And the coefficient of variation measured is all higher than 4%, illustrate that the kit repeatability is bad, performance is unstable, measurement is inaccurate.
8. preparing the sample that a apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 8
7 times, testing result is as shown in table 8.
Table 8：2-8 DEG C preserves 1 month, 3 months, 12 months measurement results
From table 8, embodiment 8 deviates actual value in the apo E concentration that 1 month, 3 months, 12 months measure,
And the coefficient of variation measured is all higher than 5%, illustrate that the kit repeatability is bad, performance is unstable, measurement is inaccurate.
It should be understood that the present invention disclosed is not limited only to specific method, scheme and the material of description, because these
Equal alterable.It will also be understood that purpose of the terminology used here just for the sake of the specific embodiment scheme of description, rather than
It is intended to limit the scope of the present invention, the scope of the present invention is limited solely by appended claim.
Those skilled in the art, which will also be appreciated that or be able to confirm that, uses no more than normal experiment, institute herein
The many equivalents for the specific embodiment of the invention stated.These equivalents are also contained in appended claim.
Claims (10)
- A kind of 1. apolipoprotein E detection kit, it is characterised in that：The kit includes reagent R1 and reagent R2, the examination Included in agent R1 in Nonidet P40 1-51g/L, the reagent R2 comprising Nonidet P40 1-51g/L, magnesium Salt 0.01-16.5g/L, calcium acetate 0.01-20g/L and antibody 10-1000mg/L.
- 2. apolipoprotein E detection kit according to claim 1, it is characterised in that：Ethylphenyl in the reagent R1 Polyethylene glycol is 5-40g/L, preferably 30g/L；Nonidet P40 is 4-41g/L in the reagent R2, is preferably 32g/L。
- 3. apolipoprotein E detection kit according to claim 1 or 2, it is characterised in that：The magnesium salts is 0.05- 10g/L, preferably 5g/L；The magnesium salts is preferably the one or more in magnesium sulfate, magnesium chloride or magnesium acetate.
- 4. according to the apolipoprotein E detection kit described in claim any one of 1-3, it is characterised in that：The calcium acetate is 0.03-16.5g/L, preferably 9g/L.
- 5. according to the apolipoprotein E detection kit described in claim any one of 1-4, it is characterised in that：In the reagent R1 Also include buffer solution, inorganic salts, preservative and aggregation；Preferably, buffer solution 20-150mmol/ is also included in the reagent R1 L, inorganic salts 1-30g/L, preservative 0.5-1g/L and aggregation 1-60g/L.
- 6. according to the apolipoprotein E detection kit described in claim any one of 1-5, it is characterised in that：In the reagent R2 Also include buffer solution, inorganic salts and preservative, it is preferable that also comprising buffer solution 20-100mmol/L, inorganic in the reagent R2 Salt 1-30g/L and preservative 0.5-1g/L.
- 7. according to the apolipoprotein E detection kit described in claim any one of 1-6, it is characterised in that：The kit bag Containing calibration object, the calibration object is calibrated using multiple spot, and the calibration object includes buffer solution 20-100mmol/L, inorganic salts 1-30g/ L, preservative 0.1-2g/L, glucan 0.3-100g/L, trehalose 1-100g/L, sucrose 1-100g/L, bovine serum albumin(BSA) 1- 100g/L and antigen.
- A kind of 8. apolipoprotein E detection kit, it is characterised in that：The kit includes reagent R1 and reagent R2, the examination Buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/ are included in agent R1 L, Nonidet P40 1-51g/L；Buffer solution 20-100mmol/L, inorganic salts 1-30g/L are included in the reagent R2, is prevented Rotten agent 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 1-51g/L, magnesium sulfate 0.01-16.5g/L, acetic acid Calcium 0.01-20g/L；Preferably,Buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, polyethylene glycol are included in the reagent R1 6000 1-60g/L, Nonidet P40 5-40g/L；Comprising buffer solution 20-100mmol/L, inorganic in the reagent R2 Salt 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 4-41g/L, magnesium sulfate 0.05- 10g/L, calcium acetate 0.03-16.5g/L；It is highly preferred thatBuffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, polyethylene glycol are included in the reagent R1 6000 1-60g/L, Nonidet P40 30g/L；Buffer solution 20-100mmol/L, inorganic salts are included in the reagent R2 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 32g/L, magnesium sulfate 5g/L, acetic acid Calcium 9g/L.
- 9. according to the apolipoprotein E detection kit described in claim any one of 1-8, it is characterised in that：The antibody is preferably goat-anti people, rabbit-anti people, horse is anti-human, mouse is anti-human or other animal anti-human antibodies；The buffer solution is preferably that acetate buffer, ammonium chloride buffer, phosphate buffer, TRIS buffer solutions, boric acid delay One or more in fliud flushing, glycine buffer, CAPSO, MOPS or Hepes buffer solution；The inorganic salts are preferably chlorination One or both of sodium, potassium chloride；The preservative is preferably Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate or ethyl mercury thiosulfuric acid One or more in sodium；The aggregation is preferably in polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or PEG 8000 It is one or more of.
- 10. a kind of detection method using above-mentioned apolipoprotein E detection kit, comprise the following steps：(1) add reagent R1 into testing sample to mix, testing sample and reagent R1 are (1-9) by volume：300 add, and 37 DEG C be incubated, under certain wavelength, read absorbance A 1；(2) add reagent R2 into the mixed liquor of step (1) to mix, reagent R2 and reagent R1 is 1 by volume：(1-6) is added, 37 DEG C of incubations, under certain wavelength, read absorbance A 2；(3) absorbance △ A, △ A=A2-A1 are drawn；(4) use calibration object on automatic clinical chemistry analyzer by built-in curve matching model fitting standard curve, according to suction Luminosity calculates the content of apo E in testing sample automatically.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112240931A (en) * | 2020-09-29 | 2021-01-19 | 中生北控生物科技股份有限公司 | Immunity turbidimetry kit |
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| CN104198730A (en) * | 2014-08-28 | 2014-12-10 | 宁波瑞源生物科技有限公司 | Apolipoprotein E detection kit |
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| CN1606452A (en) * | 2001-12-21 | 2005-04-13 | 诺沃娜第克公司 | Liquid composition of modified factor vii polypeptides |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112240931A (en) * | 2020-09-29 | 2021-01-19 | 中生北控生物科技股份有限公司 | Immunity turbidimetry kit |
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