CN107478846B - A kind of immunoglobulin G detection reagent box and detection method - Google Patents
A kind of immunoglobulin G detection reagent box and detection method Download PDFInfo
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- CN107478846B CN107478846B CN201710680451.8A CN201710680451A CN107478846B CN 107478846 B CN107478846 B CN 107478846B CN 201710680451 A CN201710680451 A CN 201710680451A CN 107478846 B CN107478846 B CN 107478846B
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- 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
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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- G01N21/82—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a precipitate or turbidity
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Abstract
The present invention provides a kind of Immunoturbidimetric kit, it includes reagent R1 and reagent R2, wherein includes Nonidet P40 in reagent R1, includes Nonidet P40, magnesium salts, calcium acetate and antibody in reagent R2.Kit antibody performance of the invention is good, reproducible, reagent testing result is accurate, can satisfy requirement.
Description
Technical field
The present invention relates to medical immunology in-vitro diagnosis fields, and in particular to a kind of immunoglobulin G detection reagent box and inspection
Survey method.
Background technique
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 scattering of measurement light caused by aggregate, 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, the specificity that there is immunoturbidimetry immunology antigen, antibody to combine, and have
Biotrepy feature can detect in body fluid, the micro test substance especially in blood in automation biochemical instruments,
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.The basic principle is that: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 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, Ji Keji
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 rouge in full-automatic biochemical measurement
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 biggish 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 effect is influenced, is affected to patient, requirement is not able to satisfy.
Summary of the invention
To solve the above problems, the invention discloses a kind of immunoglobulin G detection reagent box and detection method, reagent are steady
It is qualitative it is good, homogeneity is good, testing result accuracy is good, easy to operate, easy to promote and utilize.
For achieving the above object, the present invention provides following technical scheme:
We's invention provides a kind of immunoglobulin G detection reagent box, wherein including reagent R1 and reagent R2, the examination
Include Nonidet P40 0.9-51g/L in agent R1, includes Nonidet P40 0.9-51g/ in the reagent R2
L, magnesium salts 0.01-15g/L, calcium acetate 0.01-22g/L and antibody 10-1000mg/L.
A kind of immunoglobulin G detection reagent box, Nonidet P40 is 4.5-41g/L in the reagent R1, excellent
It is selected as 26g/L;Nonidet P40 is 5-40g/L, preferably 34g/L in the reagent R2.
A kind of immunoglobulin G detection reagent box, the magnesium salts are 0.05-10g/L, preferably 7g/L;The magnesium salts is excellent
It is selected as one of magnesium sulfate, magnesium chloride or magnesium acetate or a variety of.
A kind of immunoglobulin G detection reagent box, the calcium acetate are 0.05-16g/L, preferably 10g/L.
Wherein,
It also include buffer, inorganic salts, preservative and aggregation in the reagent R1;Preferably, in the reagent R1 also
Include buffer 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L and aggregation 1-60g/L.
It also include buffer, inorganic salts and preservative 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 immunoglobulin G detection reagent box, wherein also including calibration object, the calibration object is calibrated using multiple spot, institute
State calibration object include buffer 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 immunoglobulin G detection reagent box, wherein comprising including buffering in reagent R1 and reagent R2, the reagent R1
Liquid 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/L, the poly- second of ethylphenyl
It include buffer 20-100mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/ in glycol 0.9-51g/L, the reagent R2
L, antibody 10-1000mg/L, Nonidet P40 0.9-51g/L, magnesium sulfate 0.01-15g/L, calcium acetate 0.01-22g/
L;
Preferably,
It include buffer 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, poly- second in the reagent R1
It include buffer 20-100mmol/ in 6000 1-60g/L of glycol, Nonidet P40 4.5-41g/L, the reagent R2
L, inorganic salts 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 5-40g/L, magnesium sulfate
0.05-10g/L, calcium acetate 0.05-16g/L;
It is highly preferred that
It include buffer 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, poly- second in the reagent R1
It include buffer 20-100mmol/L, nothing in 6000 1-60g/L of glycol, Nonidet P40 26g/L, the reagent R2
Machine salt 1-30g/L, preservative 0.5-1g/L, antibody 10-1000mg/L, Nonidet P40 34g/L, magnesium sulfate 7g/L,
Calcium acetate 10g/L.
A kind of immunoglobulin G detection reagent box,
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 be acetate buffer, ammonium chloride buffer, phosphate buffer, TRIS buffer,
One or more of borate buffer, glycine buffer, CAPSO, MOPS or Hepes buffer;
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 immunoglobulin G detection reagent box, including walks as follows
Suddenly:
(1) reagent R1 is added into sample to be tested to mix, sample to be tested and reagent R1 are (1-9) by volume:300 add
Enter, 37 DEG C of incubations read absorbance A 1 under certain wavelength;
(2) reagent R2 is added into the mixed liquor of step (1) to mix, reagent R2 and reagent R1 are 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 immunoglobulin G 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 immunoglobulin G detection reagent box and detection method:
By adopting the above-described technical solution, the beneficial effects of the invention are as follows:
Immunoglobulin G detection reagent box provided by the invention and detection method, reagent stability is good, and homogeneity is good, resists
Body can stablize preservation, be not in deposited phenomenon, and antibody titer is high, performance is good, and does not have filtration step, easy to operate,
Low in cost, 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, below 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 immunoglobulin G detection reagent box of embodiment
Reagent R1:
TRIS buffer | 20mmol/L |
Sodium chloride | 1g/L |
Sodium azide | 0.5g/L |
Macrogol 6000 | 1g/L |
Nonidet P40 | 0.9g/L |
Reagent R2:
TRIS buffer | 20mmol/L |
Sodium chloride | 1g/L |
Sodium azide | 0.5g/L |
Goat anti-human immunoglobulin's G antibody | 10mg/L |
Nonidet P40 | 0.9g/L |
Magnesium sulfate | 0.01g/L |
Calcium acetate | 0.01g/L |
Calibration object:
Immunoglobulin G antigen standard dilutions (20mmol/L phosphate buffer, 2g/L sodium chloride, 0.2g/L
Sodium azide, 0.3g/L glucan, 1g/L trehalose, 2g/L sucrose, 2g/L bovine serum albumin(BSA)) dissolution, with commercially available contrast agents
It detects and adjusts to 120mg/L, packing is stored in -20 DEG C.Various concentration is diluted to using preceding taking-up, and with standard dilutions
Immunoglobulin G standard items (immunoglobulin G antigen concentration:0mg/L,10mg/L,20mg/L,50mg/L,70mg/L).So
Afterwards with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
2 immunoglobulin G detection reagent box of embodiment
Reagent R1:
Phosphate buffer | 40mmol/L |
Potassium chloride | 7g/L |
Phenol | 0.7g/L |
Macrogol 6000 | 30g/L |
Nonidet P40 | 51g/L |
Reagent R2:
Phosphate buffer | 40mmol/L |
Potassium chloride | 5g/L |
Phenol | 0.7g/L |
Goat anti-human immunoglobulin's G antibody | 50mg/L |
Nonidet P40 | 51g/L |
Magnesium sulfate | 15g/L |
Calcium acetate | 22g/L |
Calibration object:
Immunoglobulin G antigen standard dilutions (50mmol/L glycine buffer, 10g/L sodium chloride, 0.5g/L
Sodium azide, 20g/L glucan, 25g/L trehalose, 30g/L sucrose, 20g/L bovine serum albumin(BSA)) dissolution, is tried with commercially available control
Agent is detected and is adjusted to 100mg/L, and packing is stored in -20 DEG C.Using preceding taking-up, and it is diluted to standard dilutions different dense
Immunoglobulin G standard items (the immunoglobulin G antigen concentration of degree:0mg/L,10mg/L,30mg/L,50mg/L,70mg/L).
Then with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
3 immunoglobulin G detection reagent box of embodiment
Reagent R1:
Acetate buffer | 80mmol/L |
Potassium chloride | 20g/L |
Sodium azide | 0.8g/L |
Macrogol 6000 | 45g/L |
Nonidet P40 | 4.5g/L |
Reagent R2:
Calibration object:
(70mmol/L TRIS buffer, 25g/L sodium chloride, 1g/L are folded with standard dilutions for immunoglobulin G antigen
Nitrogen sodium, 55g/L glucan, 50g/L trehalose, 60g/L sucrose, 75g/L bovine serum albumin(BSA)) dissolution, with commercially available contrast agents
It detects and adjusts to 200mg/L, packing is stored in -20 DEG C.Various concentration is diluted to using preceding taking-up, and with standard dilutions
Immunoglobulin G standard items (immunoglobulin G antigen concentration:2mg/L,10mg/L,40mg/L,60mg/L,80mg/L).So
Afterwards with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
4 immunoglobulin G detection reagent box of embodiment
Reagent R1:
MOPS buffer | 150mmol/L |
Sodium chloride | 30g/L |
Sodium azide | 1g/L |
Macrogol 6000 | 55g/L |
Nonidet P40 | 41g/L |
Reagent R2:
MOPS buffer | 100mmol/L |
Sodium chloride | 30g/L |
Sodium azide | 1g/L |
Goat anti-human immunoglobulin's G antibody | 600mg/L |
Nonidet P40 | 40g/L |
Magnesium sulfate | 10g/L |
Calcium acetate | 16g/L |
Calibration object:
(100mmol/L TRIS buffer, 15g/L sodium chloride, 1g/L are folded with standard dilutions for immunoglobulin G antigen
Nitrogen sodium, 10g/L glucan, 100g/L trehalose, 95g/L sucrose, 100g/L bovine serum albumin(BSA)) dissolution, is tried with commercially available control
Agent is detected and is adjusted to 160mg/L, and packing is stored in -20 DEG C.Using preceding taking-up, and it is diluted to standard dilutions different dense
Immunoglobulin G standard items (the immunoglobulin G antigen concentration of degree:0mg/L,20mg/L,40mg/L,70mg/L,100mg/
L).Then with 0.45 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
5 immunoglobulin G detection reagent box of embodiment
Reagent R1:
MOPS buffer | 150mmol/L |
Sodium chloride | 30g/L |
Sodium azide | 1g/L |
Macrogol 6000 | 55g/L |
Nonidet P40 | 26g/L |
Reagent R2:
MOPS buffer | 100mmol/L |
Sodium chloride | 30g/L |
Sodium azide | 1g/L |
Goat anti-human immunoglobulin's G antibody | 100mg/L |
Nonidet P40 | 34g/L |
Magnesium sulfate | 7g/L |
Calcium acetate | 10g/L |
Calibration object:
Immunoglobulin G antigen standard dilutions (100mmol/L TRIS buffer, 30g/L sodium chloride, 0.5g/L
Sodium azide, 90g/L glucan, 100g/L trehalose, 100g/L sucrose, 100g/L bovine serum albumin(BSA)) dissolution, with commercially available control
Reagent is detected and is adjusted to 120mg/L, and packing is stored in -20 DEG C.Difference is diluted to using preceding taking-up, and with standard dilutions
Immunoglobulin G standard items (the immunoglobulin G antigen concentration of concentration: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 immunoglobulin G detection reagent box of embodiment
Reagent R1:
TRIS buffer | 20mmol/L |
Sodium chloride | 1g/L |
Sodium azide | 0.5g/L |
Macrogol 6000 | 1g/L |
Nonidet P40 | 0.9g/L |
Reagent R2:
Calibration object:
Immunoglobulin G antigen standard dilutions (25mmol/L phosphate buffer, 2g/L sodium chloride, 0.2g/L
Sodium azide, 0.3g/L glucan, 1g/L trehalose, 3g/L sucrose, 3g/L bovine serum albumin(BSA)) dissolution, with commercially available contrast agents
It detects and adjusts to 120mg/L, packing is stored in -20 DEG C.Various concentration is diluted to using preceding taking-up, and with standard dilutions
Immunoglobulin G standard items (immunoglobulin G antigen concentration:0mg/L,10mg/L,20mg/L,50mg/L,70mg/L).So
Afterwards with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
7 immunoglobulin G detection reagent box of embodiment
Reagent R1:
TRIS buffer | 20mmol/L |
Sodium chloride | 1g/L |
Sodium azide | 0.5g/L |
Macrogol 6000 | 1g/L |
Nonidet P40 | 0.9g/L |
Reagent R2:
TRIS buffer | 20mmol/L |
Sodium chloride | 1g/L |
Sodium azide | 0.5g/L |
Goat anti-human immunoglobulin's G antibody | 10mg/L |
Nonidet P40 | 0.9g/L |
Calcium acetate | 0.01g/L |
Calibration object:
Immunoglobulin G antigen standard dilutions (20mmol/L phosphate buffer, 2g/L sodium chloride, 0.2g/L
Sodium azide, 0.3g/L glucan, 1g/L trehalose, 2g/L sucrose, 2g/L bovine serum albumin(BSA)) dissolution, with commercially available contrast agents
It detects and adjusts to 120mg/L, packing is stored in -20 DEG C.Various concentration is diluted to using preceding taking-up, and with standard dilutions
Immunoglobulin G standard items (immunoglobulin G antigen concentration:0mg/L,10mg/L,20mg/L,50mg/L,70mg/L).So
Afterwards with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
8 immunoglobulin G detection reagent box of embodiment
Reagent R1:
TRIS buffer | 20mmol/L |
Sodium chloride | 1g/L |
Sodium azide | 0.5g/L |
Macrogol 6000 | 1g/L |
Polysorbas20 | 0.9g/L |
Reagent R2:
TRIS buffer | 20mmol/L |
Sodium chloride | 1g/L |
Sodium azide | 0.5g/L |
Goat anti-human immunoglobulin's G antibody | 10mg/L |
Polysorbas20 | 0.9g/L |
Calibration object:
Immunoglobulin G antigen standard dilutions (20mmol/L phosphate buffer, 2g/L sodium chloride, 0.2g/L
Sodium azide, 0.3g/L glucan, 1g/L trehalose, 2g/L sucrose, 2g/L bovine serum albumin(BSA)) dissolution, with commercially available contrast agents
It detects and adjusts to 120mg/L, packing is stored in -20 DEG C.Various concentration is diluted to using preceding taking-up, and with standard dilutions
Immunoglobulin G standard items (immunoglobulin G antigen concentration:0mg/L,10mg/L,20mg/L,50mg/L,70mg/L).So
Afterwards with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
Different embodiment testing results compare, wherein CV value=STDEV (1-7)/mean value:
1. preparing the sample that a IgG density is 40mg/L, sample is repeated to examine with the kit of embodiment 1
It surveys 7 times, testing result is as shown in table 1.
Table 1:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
Seen from table 1, embodiment 1 is in 1 month, 3 months, 12 months IgG densities measured close to true
Value, and the coefficient of variation (CV value) measured is respectively less than 2%, illustrates that kit of the invention is reproducible, performance is stablized, measurement
Accurately.
2. preparing the sample that a IgG density is 1.0mg/L, sample is repeated with the kit of embodiment 2
Detection 7 times, testing result is as shown in table 2.
Table 2:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
As can be seen from Table 2, embodiment 2 is in 1 month, 3 months, 12 months IgG densities measured close to true
Value, and the coefficient of variation measured is respectively less than 2%, illustrates that kit of the invention is reproducible, performance is stablized, measurement is accurate.
3. preparing the sample that a IgG density is 25mg/L, sample is repeated to examine with the kit of embodiment 3
It surveys 7 times, testing result is as shown in table 3.
Table 3:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
Seen from table 3, embodiment 3 is in 1 month, 3 months, 12 months IgG densities measured close to true
Value, and the coefficient of variation measured is respectively less than 1%, illustrates that kit of the invention is reproducible, performance is stablized, measurement is accurate.
4. preparing the sample that a IgG density is 70mg/L, sample is repeated to examine with the kit of embodiment 4
It surveys 7 times, testing result is as shown in table 4.
Table 4:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
By table 4 as it can be seen that embodiment 4 approached really in 1 month, 3 months, 12 months IgG densities measured
Value, and the coefficient of variation measured is respectively less than 1%, illustrates that kit of the invention is reproducible, performance is stablized, measurement is accurate.
5. preparing the sample that a IgG density is 25mg/L, sample is repeated to examine with the kit of embodiment 5
It surveys 7 times, testing result is as shown in table 5.
Table 5:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
By table 5 as it can be seen that embodiment 51 month, 3 months, the IgG density that measures for 12 months be true value,
And the coefficient of variation measured is 0, illustrates that kit of the invention is reproducible, performance is stablized, measurement is accurate.
6. preparing the sample that a IgG density is 40mg/L, sample is repeated to examine with the kit of embodiment 6
It surveys 7 times, testing result is as shown in table 6.
Table 6:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
By table 6 as it can be seen that embodiment 6 deviateed really in 1 month, 3 months, 12 months IgG densities measured
Value, and the coefficient of variation measured is all larger than 4%, illustrates that the kit repeatability is bad, performance is unstable, measurement inaccuracy.
7. preparing the sample that a IgG density is 40mg/L, sample is repeated to examine with the kit of embodiment 7
It surveys 7 times, testing result is as shown in table 7.
Table 7:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
By table 7 as it can be seen that embodiment 7 deviateed really in 1 month, 3 months, 12 months IgG densities measured
Value, and the coefficient of variation measured is all larger than 4%, illustrates that the kit repeatability is bad, performance is unstable, measurement inaccuracy.
8. preparing the sample that a IgG density is 40mg/L, sample is repeated to examine with the kit of embodiment 8
It surveys 7 times, testing result is as shown in table 8.
Table 8:2-8 DEG C of preservation 1 month, 3 months, 12 months measurement results
By table 8 as it can be seen that embodiment 8 deviateed really in 1 month, 3 months, 12 months IgG densities measured
Value, and the coefficient of variation measured is all larger than 5%, illustrates that the kit repeatability is bad, performance is unstable, measurement inaccuracy.
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
The many equivalents for the specific embodiment of the invention stated.These equivalents are also contained in the attached claims.
Claims (13)
1. a kind of immunoglobulin G detection reagent box, it is characterised in that:The kit includes reagent R1 and reagent R2, described
Include Nonidet P40 0.9-51g/L in reagent R1, includes Nonidet P40 0.9- in the reagent R2
51g/L, magnesium salts 0.01-15g/L, calcium acetate 0.01-22g/L and antibody 10-600mg/L;
It also include buffer 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L and aggregation in the reagent R1
1-60g/L;The aggregation is the substance for promoting antigen-antibody agglutination in the reaction;
It also include buffer 20-100mmol/L, inorganic salts 1-30g/L and preservative 0.5-1g/L in the reagent R2;
The antibody is goat-anti people, rabbit-anti people, horse is anti-human, mouse is anti-human or other animal anti-human antibodies;
In reagent R1 and reagent R2, the inorganic salts are one or both of sodium chloride, potassium chloride.
2. immunoglobulin G detection reagent box according to claim 1, it is characterised in that:Ethylo benzene in the reagent R1
Base polyethylene glycol is 4.5-41g/L;Nonidet P40 is 5-40g/L in the reagent R2.
3. immunoglobulin G detection reagent box according to claim 2, it is characterised in that:Ethylo benzene in the reagent R1
Base polyethylene glycol is 26g/L;Nonidet P40 is 34g/L in the reagent R2.
4. immunoglobulin G detection reagent box according to claim 1, it is characterised in that:The magnesium salts is 7g/L;It is described
Magnesium salts is preferably one of magnesium sulfate, magnesium chloride or magnesium acetate or a variety of.
5. immunoglobulin G detection reagent box according to claim 4, it is characterised in that:The magnesium salts is 7g/L.
6. described in any item immunoglobulin G detection reagent boxes according to claim 1, it is characterised in that:The calcium acetate is
0.05-16g/L。
7. immunoglobulin G detection reagent box according to claim 6, it is characterised in that:The calcium acetate is 10g/L.
8. immunoglobulin G detection reagent box according to claim 1-7, it is characterised in that:The kit
Comprising calibration object, the calibration object is calibrated using multiple spot, and the calibration object includes buffer 20-100mmol/L, inorganic salts 1-
30g/L, preservative 0.1-1g/L, glucan 0.3-100g/L, trehalose 1-100g/L, sucrose 1-100g/L, bovine serum albumin
White 1-100g/L and immunoglobulin G antigen;The inorganic salts are one or both of sodium chloride, potassium chloride.
9. a kind of immunoglobulin G detection reagent box, it is characterised in that:The kit includes reagent R1 and reagent R2,
It include buffer 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, polyethylene glycol in the reagent R1
6000 1-60g/L, Nonidet P40 4.5-41g/L;It include buffer 20-100mmol/L, nothing in the reagent R2
Machine salt 1-30g/L, preservative 0.5-1g/L, antibody 10-600mg/L, Nonidet P40 5-40g/L, magnesium sulfate 0.05-
10g/L, calcium acetate 0.05-16g/L;The antibody is preferably goat-anti people, rabbit-anti people, horse is anti-human, mouse is anti-human or other animals are anti-
Human antibody;
In reagent R1 and reagent R2, the inorganic salts are one or both of sodium chloride, potassium chloride.
10. immunoglobulin G detection reagent box according to claim 9, it is characterised in that:Comprising slow in the reagent R1
Fliud flushing 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L, Macrogol 6000 1-60g/L, ethylphenyl are poly-
Ethylene glycol 26g/L;In the reagent R2 comprising buffer 20-100mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L,
Antibody 10-600mg/L, Nonidet P40 34g/L, magnesium sulfate 7g/L, calcium acetate 10g/L;
In reagent R1 and reagent R2, the inorganic salts are one or both of sodium chloride, potassium chloride.
11. immunoglobulin G detection reagent box according to claim 1-7, it is characterised in that:The buffer
For acetate buffer, ammonium chloride buffer, phosphate buffer, TRIS buffer, borate buffer, glycine buffer,
One or more of CAPSO, MOPS or Hepes buffer;
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 of polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or PEG 8000
Or it is several.
12. immunoglobulin G detection reagent box according to claim 8, it is characterised in that:
The buffer be acetate buffer, ammonium chloride buffer, phosphate buffer, TRIS buffer, borate buffer,
One or more of glycine buffer, CAPSO, MOPS or Hepes buffer;
The preservative is in Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate or ethyl mercury sodium thiosulfate
One or more.
13. a kind of detection method using any one of the claim 1-7 immunoglobulin G detection reagent box, including it is as follows
Step:
(1) reagent R1 is added into sample to be tested to mix, sample to be tested and reagent R1 are (1-9) by volume:300 are added, and 37
DEG C be incubated for, under certain wavelength, read absorbance A 1;
(2) reagent R2 is added into the mixed liquor of step (1) to mix, reagent R2 and reagent R1 are 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 immunoglobulin G in sample to be tested automatically.
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