CN107478853B - A kind of apolipoprotein B detection kit and detection method - Google Patents
A kind of apolipoprotein B detection kit and detection method Download PDFInfo
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Abstract
The present invention provides a kind of apolipoprotein B detection kit, it includes reagent R1 and reagent R2, comprising including 0.9 52g/L of Nonidet P40,0.01 16g/L of magnesium salts, 0.01 20g/L of calcium acetate and antibody in Nonidet P40 0.9 52g/L, reagent R2 in wherein reagent R1.The kit antibody performance of the present 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 fields, and in particular to a kind of apolipoprotein B detection kit and detection
Method.
Background technology
Turbidimetry is widely used in clinical examination work.It is immunoturbidimetry that application is most common now.
The immuno analytical method of early stage is all largely formation, agglutination and the generation of haemolysis by observing sediment
The presence or absence of specific protein and content in sample to be tested are analyzed with the light scattering caused by aggregate is measured, it is such as immune to expand
Scattered, immunoelectrophoresis, direct and brief introduction blood clotting, passive hemagglutination, complement fixation test etc., these detection methods are at low cost, result is easy
In judgement, technically convenient for grasping, can be widely used for detecting a plurality of types of clinical samples.But since 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 in conjunction with clinical demand
Instrument.Therefore, for immunology detection, immunoturbidimetry has the specificity that immunology antigen, antibody combine, and has
Biotrepy feature, can be in automation biochemical instruments in detection body fluid, the micro test substance especially 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 to survey
Determine method.Its basic principle is：When antigen and antibody react and suitable (the general provision antibody of ratio in special dilution system
It is excessive) when, under the action of the poly- agent of the rush of the soluble immune complex of formation in dilution system, it is precipitated, is formed micro- from liquid phase
Grain, makes reaction solution turbidity occur.When antibody concentration is fixed, the amount of the immune complex of formation with amount of antigen in sample increasing
Add and increase, the turbidity of reaction solution is consequently increased.Turbidity by measuring reaction solution is compareed with series of standards product, you can meter
Calculate the content of antigen in sample.
The various detecting instruments developed according to the basic 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 during full-automatic biochemical measures
The measuring principle of albumen, haptens and other protein；It can also be applied to microorganism detection simultaneously.By accurately to more
Kind substance is quantified, and has larger clinical meaning to the diagnosis, treatment and prognosis evaluation of many diseases.
Clinically used immunoturbidimetry because its sample dosage is few, can directly on automatic clinical chemistry analyzer batch sample
It analyzes, is easy to operate, but the reagent and method established at present all have some shortcoming and defect, are mainly manifested in：
Antibody, it is easy to appear cotton-shaped or flaky precipitate, causes antibody performance to be deteriorated during preservation, repeatability is bad,
The coefficient of variation (CV) becomes larger, and directly contributes reagent testing result inaccuracy, and increase filtration step, complicated for operation, and filters
Antibody may be removed, detection result is influenced, is affected to patient, requirement cannot be met.
Invention content
To solve the above problems, the invention discloses a kind of apolipoprotein B 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 B detection kit, wherein including reagent R1 and reagent R2, the reagent
In R1 comprising in Nonidet P40 0.9-52g/L, the reagent R2 comprising Nonidet P40 0.9-52g/L,
Magnesium salts 0.01-16g/L, calcium acetate 0.01-20g/L and antibody 10-1000mg/L.
A kind of apolipoprotein B detection kit, Nonidet P40 is 5-40g/L in the reagent R1, preferably
28g/L；Nonidet P40 is 4.5-41g/L, preferably 30g/L in the reagent R2.
A kind of apolipoprotein B detection kit, the magnesium salts are 0.05-10g/L, preferably 6g/L；The magnesium salts is preferred
It is one or more in magnesium sulfate, magnesium chloride or magnesium acetate.
A kind of apolipoprotein B detection kit, the calcium acetate are 0.05-16g/L, preferably 12g/L.
Wherein,
Also include buffer solution, inorganic salts, preservative and aggregation in the reagent R1；Preferably, in the reagent R1 also
Including buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L and aggregation 1-60g/L.
Also include buffer solution, inorganic salts and preservative in the reagent R2, it is preferable that also include buffering in the reagent R2
Liquid 20-100mmol/L, inorganic salts 1-30g/L and preservative 0.5-1g/L.
A kind of apolipoprotein B 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 B detection kit, wherein comprising including buffer solution in reagent R1 and reagent R2, the reagent R1
The poly- second of 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/L, ethylphenyl two
In alcohol 0.9-52g/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 0.9-52g/L, magnesium sulfate 0.01-16g/L, calcium acetate 0.01-20g/L；
Preferably,
Include buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, poly- second in the reagent R1
In 6000 1-60g/L of glycol, 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 4.5-41g/L, magnesium sulfate
0.05-10g/L, calcium acetate 0.05-16g/L；
It is highly preferred that
Include buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, poly- second in the reagent R1
Include buffer solution 20-100mmol/L, nothing in 6000 1-60g/L of glycol, Nonidet P40 28g/L, the reagent R2
Machine salt 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 30g/L, magnesium sulfate 6g/L,
Calcium acetate 12g/L.
A kind of apolipoprotein B 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 of 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 of sodium thiosulfate；
Preferably, the aggregation is polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or polyethylene glycol
One or more of 8000；
Present aspect additionally provides a kind of detection method using above-mentioned apolipoprotein B detection kit, includes the following steps：
(1) reagent R1 mixings are added into sample to be tested, sample to be tested is (1-9) by volume with reagent R1：300 add
Enter, 37 DEG C of incubations read absorbance A 1 under certain wavelength；
(2) reagent R2 mixings are added into the mixed liquor of step (1), reagent R2 and reagent R1 is 1 by volume：(1-6)
It is added, 37 DEG C of incubations read absorbance A 2 under certain wavelength；
(3) absorbance △ A, △ A=A2-A1 are obtained；
(4) pass through built-in curve matching model fitting standard curve, root on automatic clinical chemistry analyzer using calibration object
Calculate the content of apolipoprotein B in sample to be tested automatically according to absorbance.
Signified aggregation is the substance for promoting antigen-antibody agglutination in the reaction in the present invention.
It is as follows that the present invention provides raw material sources involved in apolipoprotein B detection kit and detection method：
By adopting the above-described technical solution, the beneficial effects of the invention are as follows：
Apolipoprotein B detection kit and detection method provided by the invention, reagent stability is good, and homogeneity is good, antibody
Preservation can be stablized, be not in deposited phenomenon, antibody titer is high, performance is good, and does not have filtration step, easy to operate, at
This is cheap, and testing result is accurate, reproducible, has wider versatility.
Embodiment
In order to make those skilled in the art more fully understand the technical solution in the application, with reference to embodiment to this hair
It is bright to be described further, it is clear that described embodiments are only a part of embodiments of the present application, rather than whole implementation
Example.Based on the embodiment in the application, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present application.1 apolipoprotein B detection kit of embodiment
Reagent R1：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 0.9g/L |
Reagent R2：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein b antibody | 10mg/L |
| Nonidet P40 | 0.9g/L |
| Magnesium sulfate | 0.01g/L |
| Calcium acetate | 0.01g/L |
Calibration object：
ApoB antigens with standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.1g/L Sodium azides,
0.4g/L glucans, 1g/L trehaloses, 1g/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, standard dilutions is used in combination to be diluted to the ApoB marks of various concentration
Quasi- product (ApoB antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then it is removed with 0.65 μm of membrane filtration
Bacterium places 2~8 DEG C of preservations.2 apolipoprotein B detection kit of embodiment
Reagent R1：
| TRIS buffer solutions | 60mmol/L |
| Potassium chloride | 6g/L |
| Phenol | 0.7g/L |
| Macrogol 6000 | 25g/L |
| Nonidet P40 | 52g/L |
Reagent R2：
| TRIS buffer solutions | 60mmol/L |
| Potassium chloride | 6g/L |
| Phenol | 0.7g/L |
| Goat-anti human apolipoprotein b antibody | 50mg/L |
| Nonidet P40 | 52g/L |
| Magnesium sulfate | 16g/L |
| Calcium acetate | 20g/L |
Calibration object：
ApoB antigens with standard dilutions (40mmol/LTRIS buffer solutions, 10g/L sodium chloride, 0.1g/L Sodium azides,
50g/L glucans, 25g/L trehaloses, 30g/L sucrose, 20g/L bovine serum albumin(BSA)s) dissolving, simultaneously with the detection of commercially available contrast agents
It adjusts to 100mg/L, packing is stored in -20 DEG C.Using preceding taking-up, standard dilutions is used in combination to be diluted to the ApoB of various concentration
Standard items (ApoB antigen concentrations：0mg/L、10mg/L、30mg/L、50mg/L、70mg/L).Then with 0.65 μm of membrane filtration
2~8 DEG C of preservations are placed in degerming.
3 apolipoprotein B detection kit of embodiment
Reagent R1：
| Acetate buffer | 80mmol/L |
| Potassium chloride | 20g/L |
| Sodium azide | 0.8g/L |
| Macrogol 6000 | 50g/L |
| Nonidet P40 | 5g/L |
Reagent R2：
| Acetate buffer | 80mmol/L |
| Potassium chloride | 20g/L |
| Sodium azide | 0.8g/L |
| Goat-anti human apolipoprotein b antibody | 300mg/L |
| Nonidet P40 | 4.5g/L |
| Magnesium sulfate | 0.05g/L |
| Calcium acetate | 0.05g/L |
Calibration object：
ApoB antigens with standard dilutions (80mmol/L acetate buffers, 20g/L sodium chloride, 0.8g/L Sodium azides,
70g/L glucans, 60g/L trehaloses, 60g/L sucrose, 70g/L bovine serum albumin(BSA)s) dissolving, simultaneously with the detection of commercially available contrast agents
It adjusts to 200mg/L, packing is stored in -20 DEG C.Using preceding taking-up, standard dilutions is used in combination to be diluted to the ApoB of various concentration
Standard items (ApoB antigen concentrations：2mg/L、10mg/L、40mg/L、60mg/L、80mg/L).Then with 0.65 μm of membrane filtration
2~8 DEG C of preservations are placed in degerming.
4 apolipoprotein B detection kit of embodiment
Reagent R1：
| MOPS buffer solutions | 120mmol/L |
| Sodium chloride | 20g/L |
| Sodium azide | 1g/L |
| Macrogol 6000 | 60g/L |
| Nonidet P40 | 40g/L |
Reagent R2：
| MOPS buffer solutions | 100mmol/L |
| Sodium chloride | 20g/L |
| Sodium azide | 1g/L |
| Goat-anti human apolipoprotein b antibody | 600mg/L |
| Nonidet P40 | 41g/L |
| Magnesium sulfate | 10g/L |
| Calcium acetate | 16g/L |
Calibration object：
ApoB antigens with standard dilutions (100mmol/L MOPS buffer solutions, 25g/L sodium chloride, 2g/L Sodium azides,
20g/L glucans, 100g/L trehaloses, 100g/L sucrose, 100g/L bovine serum albumin(BSA)s) dissolving, it is examined with commercially available contrast agents
It surveys and adjusts to 160mg/L, packing is stored in -20 DEG C.Using preceding taking-up, standard dilutions is used in combination to be diluted to various concentration
ApoB standard items (ApoB antigen concentrations：0mg/L、20mg/L、40mg/L、70mg/L、100mg/L).Then with 0.45 μm of filter
2~8 DEG C of preservations are placed in membrane filtration degerming.
5 apolipoprotein B detection kit of embodiment
Reagent R1：
| MOPS buffer solutions | 150mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Macrogol 6000 | 60g/L |
| Nonidet P40 | 28g/L |
Reagent R2：
| MOPS buffer solutions | 100mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Goat-anti human apolipoprotein b antibody | 1000mg/L |
| Nonidet P40 | 30g/L |
| Magnesium sulfate | 6g/L |
| Calcium acetate | 12g/L |
Calibration object：
ApoB antigens with standard dilutions (100mmol/L MOPS buffer solutions, 30g/L sodium chloride, 1g/L Sodium azides,
90g/L glucans, 100g/L trehaloses, 100g/L sucrose, 100g/L bovine serum albumin(BSA)s) dissolving, it is examined with commercially available contrast agents
It surveys and adjusts to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, standard dilutions is used in combination to be diluted to various concentration
ApoB standard items (ApoB antigen concentrations：0mg/L、20mg/L、50mg/L、80mg/L、100mg/L).Then with 0.65 μm of filter
2~8 DEG C of preservations are placed in membrane filtration degerming.
6 apolipoprotein B detection kit of embodiment
Reagent R1：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 0.9g/L |
Reagent R2：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein b antibody | 10mg/L |
| Nonidet P40 | 0.9g/L |
| Magnesium sulfate | 0.01g/L |
Calibration object：
ApoB antigens with standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.1g/L Sodium azides,
0.4g/L glucans, 1g/L trehaloses, 1g/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, standard dilutions is used in combination to be diluted to the ApoB marks of various concentration
Quasi- product (ApoB antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then it is removed with 0.65 μm of membrane filtration
Bacterium places 2~8 DEG C of preservations.
7 apolipoprotein B detection kit of embodiment
Reagent R1：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 0.9g/L |
Reagent R2：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein b antibody | 10mg/L |
| Nonidet P40 | 0.9g/L |
| Calcium acetate | 0.01g/L |
Calibration object：
ApoB antigens with standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.1g/L Sodium azides,
0.4g/L glucans, 1g/L trehaloses, 1g/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, standard dilutions is used in combination to be diluted to the ApoB marks of various concentration
Quasi- product (ApoB antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then it is removed with 0.65 μm of membrane filtration
Bacterium places 2~8 DEG C of preservations.
8 apolipoprotein B detection kit of embodiment
Reagent R1：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 0.9g/L |
Reagent R2：
| Sulfate buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein b antibody | 10mg/L |
| Polysorbas20 | 0.9g/L |
Calibration object：
ApoB antigens with standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.1g/L Sodium azides,
0.4g/L glucans, 1g/L trehaloses, 1g/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, standard dilutions is used in combination to be diluted to the ApoB marks of various concentration
Quasi- product (ApoB antigen concentrations：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then it is removed with 0.65 μm of membrane filtration
Bacterium places 2~8 DEG C of preservations.
Different embodiment testing results compare, wherein CV values=STDEV (1-7)/mean value.
1. preparing the sample of a concentration of 40mg/L of a apolipoprotein B, 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
By table 1 as it can be seen that the apolipoprotein B concentration that measures 1 month, 3 months, 12 months of embodiment 1 is close to actual value,
And the coefficient of variation (CV values) measured) be respectively less than 2%, illustrate the present invention kit is reproducible, performance is stablized, measure accurate
Really.
2. preparing the sample of a concentration of 1mg/L of a apolipoprotein B, 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
As can be seen from Table 2, the apolipoprotein B concentration that measures 1 month, 3 months, 12 months of embodiment 2 is close to actual value,
And the coefficient of variation measured is respectively less than 2%, illustrate the present invention kit is reproducible, performance is stablized, it is accurate to measure.
3. preparing the sample of a concentration of 25mg/L of a apolipoprotein B, 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
By table 3 as it can be seen that the apolipoprotein B concentration that measures 1 month, 3 months, 12 months of embodiment 3 is close to actual value,
And the coefficient of variation measured is respectively less than 1%, illustrate the present invention kit is reproducible, performance is stablized, it is accurate to measure.
4. preparing the sample of a concentration of 70mg/L of a apolipoprotein B, 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
By table 4 as it can be seen that the apolipoprotein B concentration that measures 1 month, 3 months, 12 months of embodiment 4 is close to actual value,
And the coefficient of variation measured is respectively less than 1%, illustrate the present invention kit is reproducible, performance is stablized, it is accurate to measure.
5. preparing the sample of a concentration of 25mg/L of a apolipoprotein B, 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
By table 5 as it can be seen that the apolipoprotein B concentration that embodiment 5 was measured 1 month, 3 months, 12 months is actual value, and
And the coefficient of variation measured is respectively less than 0.05%, illustrate the present invention kit is reproducible, performance is stablized, it is accurate to measure.
6. preparing the sample of a concentration of 40mg/L of a apolipoprotein B, 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
By table 6 as it can be seen that the apolipoprotein B concentration that embodiment 6 was measured 1 month, 3 months, 12 months deviates actual value,
And the coefficient of variation measured is all higher than 4%, illustrates that the kit repeatability is bad, performance is unstable, measures inaccurate.
7. preparing the sample of a concentration of 40mg/L of a apolipoprotein B, 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
By table 7 as it can be seen that the apolipoprotein B concentration that embodiment 7 was measured 1 month, 3 months, 12 months deviates actual value,
And the coefficient of variation measured is all higher than 4%, illustrates that the kit repeatability is bad, performance is unstable, measures inaccurate.
8. preparing the sample of a concentration of 40mg/L of a apolipoprotein B, 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
By table 8 as it can be seen that the apolipoprotein B concentration that embodiment 8 was measured 1 month, 3 months, 12 months deviates actual value,
And the coefficient of variation measured is all higher than 5%, illustrates that the kit repeatability is bad, performance is unstable, measures inaccurate.
It should be understood that the present invention disclosed is not limited only to specific method, scheme and the substance of description, because these
It is 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 invention, the scope of the present invention is limited solely by the attached claims.
Those skilled in the art, which will also be appreciated that or be able to confirm that, uses no more than routine experiment, institute herein
Many equivalents of the specific embodiment of the present invention stated.These equivalents are also contained in the attached claims.
Claims (15)
1. a kind of apolipoprotein B detection kit, it is characterised in that：The kit includes reagent R1 and reagent R2, the examination
Comprising including Nonidet P40 0.9-52g/ in Nonidet P40 0.9-52g/L, the reagent R2 in agent R1
L, magnesium salts 0.01-16g/L, calcium acetate 0.01-20g/L and antibody 10-1000mg/L；It also include buffering in the wherein described reagent R1
Liquid, inorganic salts, preservative and aggregation include also buffer solution, inorganic salts and preservative in the reagent R2, wherein described in R2
Inorganic salts are one or both of sodium chloride, potassium chloride；And wherein
The antibody is the anti-human apolipoprotein B antibody of animal,
The aggregation is one kind in polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or PEG 8000
Or it is several.
2. apolipoprotein B detection kit according to claim 1, it is characterised in that：Ethylphenyl in the reagent R1
Polyethylene glycol is 5-40g/L；Nonidet P40 is 4.5-41g/L in the reagent R2.
3. apolipoprotein B detection kit according to claim 2, it is characterised in that：Ethylphenyl in the reagent R1
Polyethylene glycol is 28g/L；Nonidet P40 is 30g/L in the reagent R2.
4. apolipoprotein B detection kit according to claim 1, it is characterised in that：The magnesium salts is 0.05-10g/；
The magnesium salts is one or more in magnesium sulfate, magnesium chloride or magnesium acetate.
5. apolipoprotein B detection kit according to claim 4, it is characterised in that：The magnesium salts is 6g/L.
6. apolipoprotein B detection kit according to claim 1, it is characterised in that：The calcium acetate is 0.05-16g/
L。
7. apolipoprotein B detection kit according to claim 6, it is characterised in that：The calcium acetate is 12g/L.
8. according to claim 1-7 any one of them apolipoprotein B detection kits, it is characterised in that：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.
9. according to claim 1-7 any one of them apolipoprotein B detection kits, it is characterised in that：In the reagent R2
Also include buffer solution 20-100mmol/L, inorganic salts 1-30g/L and preservative 0.5-1g/L, wherein inorganic salts described in R2 are chlorine
Change one or both of sodium, potassium chloride.
10. apolipoprotein B detection kit according to claim 1, it is characterised in that：The kit includes calibration
Product, the calibration object are calibrated using multiple spot, and the calibration object includes buffer solution 20-100mmol/L, inorganic salts 2-30g/L, anti-corrosion
Agent 0.1-2g/L, glucan 0.4-90g/L, trehalose 1-100g/L, sucrose 1-100g/L, bovine serum albumin(BSA) 2-100g/L and
Human apolipoprotein b antigen.
11. a kind of apolipoprotein B detection kit, it is characterised in that：The kit includes reagent R1 and reagent R2, the examination
Include buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/ in agent R1
L, Nonidet P40 0.9-52g/L；Include buffer solution 20-100mmol/L, sodium chloride or potassium chloride in the reagent R2
The anti-human apolipoprotein B antibody 10-1000mg/L of 1-30g/L, preservative 0.5-1g/L, animal, Nonidet P40 0.9-
52g/L, magnesium sulfate 0.01-16g/L, calcium acetate 0.01-20g/L.
12. kit according to claim 11, wherein including buffer solution 20-150mmol/L in the reagent R1, inorganic
Salt 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/L, Nonidet P40 5-40g/L；The examination
Include buffer solution 20-100mmol/L, sodium chloride or potassium chloride 1-30g/L, preservative 0.5-1g/L, the anti-human load of animal in agent R2
Lipoprotein B antibody 10-1000mg/L, Nonidet P40 4.5-41g/L, magnesium sulfate 0.05-10g/L, calcium acetate 0.05-
16g/L。
13. kit according to claim 12, wherein including buffer solution 20-150mmol/L in the reagent R1, inorganic
Salt 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/L, Nonidet P40 28g/L；The reagent
Include buffer solution 20-100mmol/L, sodium chloride or potassium chloride 1-30g/L, preservative 0.5-1g/L, the anti-human load fat of animal in R2
Protein B antibody 10-1000mg/L, Nonidet P40 30g/L, magnesium sulfate 6g/L, calcium acetate 12g/L.
14. the apolipoprotein B detection kit according to claim 1 or 11, it is characterised in that：
The antibody is goat-anti people, rabbit-anti people, horse is anti-human, mouse is anti-human or other animal anti-human antibodies；
The buffer solution be acetate buffer, ammonium chloride buffer, phosphate buffer, TRIS buffer solutions, borate buffer,
One or more of glycine buffer, CAPSO, MOPS or Hepes buffer solution；
The inorganic salts are one or both of sodium chloride, potassium chloride；
The preservative is in Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate or ethyl mercury sodium thiosulfate
One or more；
The aggregation is one kind in polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or PEG 8000
Or it is several.
15. a kind of apolipoprotein B using any one of claim 1 or 11 for non-disease diagnosis or therapeutic purposes detects examination
The detection method of agent box, includes the following steps：
(1) reagent R1 mixings are added into sample to be tested, sample to be tested is (1-9) by volume with reagent R1：300 are added, and 37
DEG C be incubated, under certain wavelength, read absorbance A 1；
(2) reagent R2 mixings are added into the mixed liquor of step (1), reagent R2 and reagent R1 is 1 by volume：(1-6) is added,
37 DEG C of incubations read absorbance A 2 under certain wavelength；
(3) absorbance △ A, △ A=A2-A1 are obtained；
(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 apolipoprotein B in sample to be tested automatically.
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| CN111024963A (en) * | 2019-12-27 | 2020-04-17 | 桂林英美特生物技术研究所 | Liquid-stable anti-freezing immunoturbidimetric apolipoprotein B single reagent |
| CN111426845B (en) * | 2020-04-07 | 2021-06-15 | 中拓生物有限公司 | Serum apolipoprotein B determination kit and preparation method and application thereof |
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