CN107490697B - A kind of kit for testing prealbumin and detection method - Google Patents
A kind of kit for testing prealbumin and detection method Download PDFInfo
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- CN107490697B CN107490697B CN201710679344.3A CN201710679344A CN107490697B CN 107490697 B CN107490697 B CN 107490697B CN 201710679344 A CN201710679344 A CN 201710679344A CN 107490697 B CN107490697 B CN 107490697B
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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
Abstract
The present invention provides a kind of Immunoturbidimetric kit, it includes reagent R1 and reagent R2, includes Nonidet P40 in wherein reagent R1, includes Nonidet P40, magnesium salts, calcium acetate and antibody in reagent R2.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 kit for testing prealbumin and detection side
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 kit for testing prealbumin 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 kit for testing prealbumin, wherein including reagent R1 and reagent R2, the reagent
Comprising including Nonidet P40 1-50g/L, magnesium salts in Nonidet P40 1-50g/L, the reagent R2 in R1
0.01-17g/L, calcium acetate 0.01-21g/L and antibody 10-1000mg/L.
A kind of kit for testing prealbumin, Nonidet P40 is 5-40g/L in the reagent R1, preferably
27g/L；Nonidet P40 is 4-43g/L, preferably 31g/L in the reagent R2.
A kind of kit for testing prealbumin, the magnesium salts are 0.05-10.5g/L, preferably 3g/L；The magnesium salts is preferred
It is one or more in magnesium sulfate, magnesium chloride or magnesium acetate.
A kind of kit for testing prealbumin, the calcium acetate are 0.05-15g/L, preferably 11g/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 kit for testing prealbumin, wherein also including calibration object, the calibration object is calibrated using multiple spot, the school
Quasi- product include buffer solution 20-100mmol/L, inorganic salts 1-30g/L, preservative 0.1-2g/L, glucan 0.3-100g/L, seaweed
Sugared 1-100g/L, sucrose 1-100g/L, bovine serum albumin(BSA) 1-100g/L and antigen.
A kind of kit for testing prealbumin, 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
Include buffer solution 20-100mmol/L in alcohol 1-50g/L, the reagent R2, inorganic salts 1-30g/L, preservative 0.5-1g/L, resist
Body 10-1000mg/L, Nonidet P40 1-50g/L, magnesium sulfate 0.01-17g/L, calcium acetate 0.01-21g/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 4-43g/L, magnesium sulfate
0.05-10.5g/L, calcium acetate 0.05-15g/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 27g/L, the reagent R2
Machine salt 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 31g/L, magnesium sulfate 3g/L,
Calcium acetate 11g/L.
A kind of kit for testing prealbumin,
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 kit for testing prealbumin, 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 prealbumin 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 kit for testing prealbumin and detection method：
By adopting the above-described technical solution, the beneficial effects of the invention are as follows：
Kit for testing prealbumin provided by the invention and detection method, reagent stability is good, and homogeneity is good, and antibody can
It is preserved with stablizing, is not in deposited phenomenon, antibody titer is high, performance is good, and does not have filtration step, easy to operate, cost
Cheap, 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 kit for testing prealbumin of embodiment
Reagent R1：
| Phosphate buffer | 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 people's prealbumin antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Magnesium sulfate | 0.01g/L |
| Calcium acetate | 0.01g/L |
Calibration object：
Prealbumin antigen standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L nitrine
Sodium, 0.3g/L glucans, 1g/L trehaloses, 2g/L sucrose, 2g/L bovine serum albumin(BSA)s) dissolving, is detected with commercially available contrast agents
And adjust to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, before being used in combination standard dilutions to be diluted to various concentration
Albumin standard (prealbumin antigen concentration：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then with 0.65 μm
Membrane filtration degerming, place 2~8 DEG C preservation.
2 kit for testing prealbumin of embodiment
Reagent R1：
Reagent R2：
| Phosphate buffer | 50mmol/L |
| Potassium chloride | 15g/L |
| Phenol | 0.6g/L |
| Goat-anti people's prealbumin antibody | 30mg/L |
| Nonidet P40 | 50g/L |
| Magnesium sulfate | 16.5g/L |
| Calcium acetate | 21g/L |
Calibration object：
(50mmol/L glycine buffers, 10g/L sodium chloride, 0.5g/L are folded with standard dilutions for prealbumin antigen
Nitrogen sodium, 20g/L glucans, 25g/L trehaloses, 30g/L sucrose, 20g/L bovine serum albumin(BSA)s) dissolving, with commercially available contrast agents
It detects and 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 various concentration
Prealbumin standard items (prealbumin antigen concentration：0mg/L、10mg/L、30mg/L、50mg/L、70mg/L).Then it uses
2~8 DEG C of preservations are placed in 0.65 μm of membrane filtration degerming.
3 kit for testing prealbumin of embodiment
Reagent R1：
| Acetate buffer | 100mmol/L |
| Potassium chloride | 20g/L |
| Sodium azide | 0.8g/L |
| Macrogol 6000 | 40g/L |
| Nonidet P40 | 5g/L |
Reagent R2：
| Acetate buffer | 60mmol/L |
| Potassium chloride | 20g/L |
| Sodium azide | 0.8g/L |
| Goat-anti people's prealbumin antibody | 300mg/L |
| Nonidet P40 | 4g/L |
| Magnesium sulfate | 0.05g/L |
| Calcium acetate | 0.05g/L |
Calibration object：
Prealbumin antigen standard dilutions (70mmol/L TRIS buffer solutions, 25g/L sodium chloride, 1g/L nitrine
Sodium, 55g/L glucans, 50g/L trehaloses, 60g/L sucrose, 75g/L bovine serum albumin(BSA)s) dissolving, it is examined with commercially available contrast agents
It surveys and 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 various concentration
Prealbumin standard items (prealbumin antigen concentration：2mg/L、10mg/L、40mg/L、60mg/L、80mg/L).Then with 0.65
μm membrane filtration degerming, place 2~8 DEG C preservation.
4 kit for testing prealbumin of embodiment
Reagent R1：
| MOPS buffer solutions | 120mmol/L |
| Sodium chloride | 20g/L |
| Sodium azide | 1g/L |
| Macrogol 6000 | 50g/L |
| Nonidet P40 | 40g/L |
Reagent R2：
| MOPS buffer solutions | 100mmol/L |
| Sodium chloride | 20g/L |
| Sodium azide | 1g/L |
| Goat-anti people's prealbumin antibody | 500mg/L |
| Nonidet P40 | 41g/L |
| Magnesium sulfate | 11g/L |
| Calcium acetate | 15g/L |
Calibration object：
Prealbumin antigen standard dilutions (100mmol/L TRIS buffer solutions, 15g/L sodium chloride, 1g/L nitrine
Sodium, 10g/L glucans, 100g/L trehaloses, 95g/L sucrose, 100g/L bovine serum albumin(BSA)s) dissolving, with commercially available contrast agents
It detects 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
Prealbumin standard items (prealbumin antigen concentration：0mg/L、20mg/L、40mg/L、70mg/L、100mg/L).Then it uses
2~8 DEG C of preservations are placed in 0.45 μm of membrane filtration degerming.
5 kit for testing prealbumin of embodiment
Reagent R1：
Reagent R2：
| MOPS buffer solutions | 70mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Goat-anti people's prealbumin antibody | 100mg/L |
| Nonidet P40 | 31g/L |
| Magnesium sulfate | 3g/L |
| Calcium acetate | 11g/L |
Calibration object：
(100mmol/L TRIS buffer solutions, 30g/L sodium chloride, 0.5g/L are folded with standard dilutions for prealbumin antigen
Nitrogen sodium, 90g/L glucans, 100g/L trehaloses, 100g/L sucrose, 100g/L bovine serum albumin(BSA)s) dissolving, is tried with commercially available control
Agent is detected and is adjusted to 120mg/L, and packing is stored in -20 DEG C.Using preceding taking-up, it is used in combination standard dilutions to be diluted to different dense
Prealbumin standard items (the prealbumin antigen concentration of degree：0mg/L、20mg/L、50mg/L、80mg/L、100mg/L).Then
With 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
6 kit for testing prealbumin of embodiment
Reagent R1：
| Phosphate buffer | 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 people's prealbumin antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Magnesium sulfate | 0.01g/L |
Calibration object：
Prealbumin antigen standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L nitrine
Sodium, 0.3g/L glucans, 1g/L trehaloses, 2g/L sucrose, 2g/L bovine serum albumin(BSA)s) dissolving, is detected with commercially available contrast agents
And adjust to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, before being used in combination standard dilutions to be diluted to various concentration
Albumin standard (prealbumin antigen concentration：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then with 0.65 μm
Membrane filtration degerming, place 2~8 DEG C preservation.
7 kit for testing prealbumin of embodiment
Reagent R1：
| Phosphate buffer | 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 people's prealbumin antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Calcium acetate | 0.01g/L |
Calibration object：
Prealbumin antigen standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L nitrine
Sodium, 0.3g/L glucans, 1g/L trehaloses, 2g/L sucrose, 2g/L bovine serum albumin(BSA)s) dissolving, is detected with commercially available contrast agents
And adjust to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, before being used in combination standard dilutions to be diluted to various concentration
Albumin standard (prealbumin antigen concentration：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then with 0.65 μm
Membrane filtration degerming, place 2~8 DEG C preservation.
8 kit for testing prealbumin of embodiment
Reagent R1：
| Phosphate buffer | 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 people's prealbumin antibody | 10mg/L |
| Polysorbas20 | 1g/L |
Calibration object：
Prealbumin antigen standard dilutions (20mmol/L phosphate buffers, 2g/L sodium chloride, 0.2g/L nitrine
Sodium, 0.3g/L glucans, 1g/L trehaloses, 2g/L sucrose, 2g/L bovine serum albumin(BSA)s) dissolving, is detected with commercially available contrast agents
And adjust to 120mg/L, packing is stored in -20 DEG C.Using preceding taking-up, before being used in combination standard dilutions to be diluted to various concentration
Albumin standard (prealbumin antigen concentration：0mg/L、10mg/L、20mg/L、50mg/L、70mg/L).Then with 0.65 μm
Membrane filtration degerming, place 2~8 DEG C preservation.
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 prealbumin, detection 7 is repeated to sample with the kit of embodiment 1
Secondary, 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 prealbumin 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 prealbumin, 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 prealbumin 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 prealbumin, detection 7 is repeated to sample with the kit of embodiment 3
Secondary, 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 prealbumin 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 60mg/L of a prealbumin, detection 7 is repeated to sample with the kit of embodiment 4
Secondary, 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 prealbumin 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 5mg/L of a prealbumin, detection 7 is repeated to sample with the kit of embodiment 5
Secondary, 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 prealbumin 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 0.5%, illustrate the present invention kit is reproducible, performance is stablized, it is accurate to measure.
6. preparing the sample of a concentration of 30mg/L of a prealbumin, detection 7 is repeated to sample with the kit of embodiment 6
Secondary, 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 prealbumin 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 5%, illustrates that the kit repeatability is bad, performance is unstable, measures inaccurate.
7. preparing the sample of a concentration of 30mg/L of a prealbumin, detection 7 is repeated to sample with the kit of embodiment 7
Secondary, 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 prealbumin 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 5%, illustrates that the kit repeatability is bad, performance is unstable, measures inaccurate.
8. preparing the sample of a concentration of 40mg/L of a prealbumin, detection 7 is repeated to sample with the kit of embodiment 8
Secondary, 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 prealbumin 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 kit for testing prealbumin, it is characterised in that：The kit includes reagent R1 and reagent R2, the reagent
Comprising including Nonidet P40 1-50g/L, magnesium salts in Nonidet P40 1-50g/L, the reagent R2 in R1
0.01-17g/L, calcium acetate 0.01-21g/L and the anti-human prealbumin antibody 10-500mg/L of animal；It is also wrapped in the reagent R1
Also include buffer solution, sodium chloride or potassium chloride and anti-corrosion containing buffer solution, inorganic salts, preservative and aggregation, in the reagent R2
Agent；And the wherein described aggregation is in polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or PEG 8000
It is one or more of.
2. kit for testing prealbumin according to claim 1, it is characterised in that：Ethylphenyl is poly- in the reagent R1
Ethylene glycol is 5-40g/L；Nonidet P40 is 4-43g/L in the reagent R2.
3. kit for testing prealbumin according to claim 2, it is characterised in that：Ethylphenyl is poly- in the reagent R1
Ethylene glycol is 27g/L；Nonidet P40 is 31g/L in the reagent R2.
4. kit for testing prealbumin according to claim 1, it is characterised in that：The magnesium salts is 0.05-10.5g/
L；The magnesium salts is one or more in magnesium sulfate, magnesium chloride or magnesium acetate.
5. kit for testing prealbumin according to claim 4, it is characterised in that：The magnesium salts is 3g/L.
6. kit for testing prealbumin according to claim 1, it is characterised in that：The calcium acetate is 0.05-15g/
L。
7. kit for testing prealbumin according to claim 6, it is characterised in that：The calcium acetate is 11g/L.
8. according to claim 1-7 any one of them kit for testing prealbumin, it is characterised in that：In the reagent R1 also
Including buffer solution 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L and aggregation 1-50g/L.
9. according to claim 1-7 any one of them kit for testing prealbumin, it is characterised in that：In the reagent R2 also
Including buffer solution 20-100mmol/L, sodium chloride or potassium chloride 1-30g/L and preservative 0.5-1g/L.
10. kit for testing prealbumin according to claim 1, it is characterised in that：The kit includes calibration object,
The calibration object is calibrated using multiple spot, and the calibration object includes buffer solution 20-100mmol/L, inorganic salts 2-30g/L, preservative
0.2-1g/L, glucan 0.3-90g/L, trehalose 1-100g/L, sucrose 2-100g/L, bovine serum albumin(BSA) 2-100g/L and preceding
Albumin antigen.
11. a kind of kit for testing prealbumin, 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 1-50g/ in agent R1
L, Nonidet P40 1-50g/L；Include buffer solution 20-100mmol/L, sodium chloride or potassium chloride 1- in the reagent R2
The anti-human prealbumin antibody 10-500mg/L of 30g/L, preservative 0.5-1g/L, animal, Nonidet P40 1-50g/L,
Magnesium sulfate 0.01-17g/L, calcium acetate 0.01-21g/L.
12. kit for testing prealbumin according to claim 11, it is characterised in that：Include buffering in the reagent R1
Liquid 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-50g/L, the poly- second of ethylphenyl
Glycol 5-40g/L；Include buffer solution 20-100mmol/L, sodium chloride or potassium chloride 1-30g/L, preservative in the reagent R2
The anti-human prealbumin antibody 10-500mg/L of 0.5-1g/L, animal, Nonidet P40 4-43g/L, magnesium sulfate 0.05-
10.5g/L, calcium acetate 0.05-15g/L.
13. kit for testing prealbumin according to claim 12, it is characterised in that：Include buffering in the reagent R1
Liquid 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-50g/L, the poly- second of ethylphenyl
Glycol 27g/L；Include buffer solution 20-100mmol/L, sodium chloride or potassium chloride 1-30g/L, preservative 0.5- in the reagent R2
The anti-human prealbumin antibody 10-500mg/L of 1g/L, animal, Nonidet P40 31g/L, magnesium sulfate 3g/L, calcium acetate
11g/L。
14. the kit for testing prealbumin 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 the anti-human prealbumin antibody of other animals；
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.
15. a kind of utilizing the kit for testing prealbumin described in claim 1 or 11 for non-disease diagnoses and treatment purpose
Detection method, include 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 Δ A, Δ 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 prealbumin in sample to be tested automatically.
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| CN108680754B (en) * | 2018-05-04 | 2021-02-26 | 湖北科技学院 | Prealbumin determination kit |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101498732A (en) * | 2008-02-03 | 2009-08-05 | 北京九强生物技术有限公司 | Improved prealbumin detection kit |
| CN104725506A (en) * | 2013-12-23 | 2015-06-24 | 上海复星医药（集团）股份有限公司 | Method for purifying human serum prealbumin polyclonal antibody by application of immunoaffinity column |
| CN105137090A (en) * | 2015-09-30 | 2015-12-09 | 山东博科生物产业有限公司 | High-accuracy prealbumin immunoturbidimetry detection kit |
| CN106501526A (en) * | 2016-10-03 | 2017-03-15 | 王贤俊 | A kind of combination directing agent optimizes latex coupled antibody detection prealbumin（PA）Test kit |
| CN106526198A (en) * | 2016-10-03 | 2017-03-22 | 王贤俊 | Combined orientation agents optimized latex coupled antibody detection method of prealbumin (PA) |
-
2017
- 2017-08-10 CN CN201710679344.3A patent/CN107490697B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101498732A (en) * | 2008-02-03 | 2009-08-05 | 北京九强生物技术有限公司 | Improved prealbumin detection kit |
| CN104725506A (en) * | 2013-12-23 | 2015-06-24 | 上海复星医药（集团）股份有限公司 | Method for purifying human serum prealbumin polyclonal antibody by application of immunoaffinity column |
| CN105137090A (en) * | 2015-09-30 | 2015-12-09 | 山东博科生物产业有限公司 | High-accuracy prealbumin immunoturbidimetry detection kit |
| CN106501526A (en) * | 2016-10-03 | 2017-03-15 | 王贤俊 | A kind of combination directing agent optimizes latex coupled antibody detection prealbumin（PA）Test kit |
| CN106526198A (en) * | 2016-10-03 | 2017-03-22 | 王贤俊 | Combined orientation agents optimized latex coupled antibody detection method of prealbumin (PA) |
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