CN107490675B - A kind of Immunoturbidimetric kit and detection method - Google Patents
A kind of Immunoturbidimetric kit and detection method Download PDFInfo
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Abstract
The present invention provides a kind of Immunoturbidimetric kit, it includes reagent R1 and reagent R2, wherein include Nonidet P40 1-50g/L in reagent R1, includes Nonidet P40 1-50g/L, magnesium salts 0.01-15g/L, calcium acetate 0.01-20g/L 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 Immunoturbidimetric kit and detection 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 Immunoturbidimetric kit and detection method, reagent stability 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 the following technical schemes are provided:
The present invention provides a kind of Immunoturbidimetric kits, wherein include reagent R1 and reagent R2,
It include Nonidet P40 1-50g/L, preferably 5-40g/L, more preferably 30g/L in the reagent R1；
It include Nonidet P40 1-50g/L, preferably 5-40g/L, more preferably 30g/L in the reagent R2；
It include magnesium salts 0.01-15g/L, preferably 0.05-10g/L, more preferably 5g/L in the reagent R2；The magnesium salts
Preferably one of magnesium sulfate, magnesium chloride or magnesium acetate or a variety of.
It include calcium acetate 0.01-20g/L, preferably 0.05-15g/L, more preferably 10g/L in shown reagent R2.
It include antibody 10-1000mg/L in the reagent R2.
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, preservative in the reagent R2；Preferably, also comprising buffering in the reagent R2
Liquid 20-100mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L.
The kit includes calibration object, and the calibration object is calibrated using multiple spot；The calibration object includes 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.
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 or 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.
Wherein, the kit for detect Apolipoprotein A1, apolipoprotein B, immunoglobulin A, immunoglobulin G,
Immunoglobulin M, Complement C4, apo E, C reactive protein, RBP ELISA, rheumatoid factor, is immunized Complement C_3
Globulin E, prealbumin or lipoprotein a.
Present aspect additionally provides a kind of detection method using above-mentioned Immunoturbidimetric kit, includes the following steps:
(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:(1-6 by volume)
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 test substance 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.
Raw material sources involved in Immunoturbidimetric kit and detection method provided by the invention are as follows:
By adopting the above-described technical solution, the beneficial effects of the present invention are:
Immunoturbidimetric kit and detection method provided by the invention, reagent stability is good, and homogeneity is good, and antibody can be steady
It is fixed to save, it is not in deposited phenomenon, antibody titer is high, performance is good, and does not have filtration step, and it is easy to operate, it is low in cost,
Testing result is accurate, reproducible, has wider versatility.
Detailed description of the invention
Fig. 1 illustrates embodiment 5 and saves 1 month result.
Fig. 2 illustrates embodiment 5 and saves 3 months results.
Fig. 3 illustrates embodiment 5 and saves 12 months results.
Fig. 4 illustrates embodiment 8 and saves 1 month result.
Fig. 5 illustrates embodiment 8 and saves 3 months results
Fig. 6 illustrates embodiment 8 and saves 12 months results.
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 apolipoprotein E detection kit of embodiment
Reagent R1:
| TRIS buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 1g/L |
Reagent R2:
| TRIS buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein E antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Magnesium sulfate | 0.01g/L |
| Calcium acetate | 0.01g/L |
Calibration object:
ApoE antigen with standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L Sodium azide,
0.3g/L glucan, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin(BSA)) dissolution, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.It is marked using preceding taking-up, and with the ApoE that standard dilutions are diluted to various concentration
Quasi- product (ApoE antigen concentration: 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 A1 detection kit of embodiment
Reagent R1:
| Phosphate buffer | 30mmol/L |
| Potassium chloride | 5g/L |
| Phenol | 0.7g/L |
| Macrogol 6000 | 20g/L |
| Nonidet P40 | 50g/L |
Reagent R2:
| Phosphate buffer | 40mmol/L |
| Potassium chloride | 5g/L |
| Phenol | 0.7g/L |
| Goat-anti human apolipoprotein A1 antibody | 50mg/L |
| Nonidet P40 | 50g/L |
| Magnesium sulfate | 15g/L |
| Calcium acetate | 20g/L |
Calibration object:
Apolipoprotein A1 antigen standard dilutions (40mmol/L glycine buffer, 10g/L sodium chloride, 0.7g/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
The Apolipoprotein A1 standard items (Apolipoprotein A1 antigen concentration: 0mg/L, 10mg/L, 30mg/L, 50mg/L, 70mg/L) of degree.So
Afterwards with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
Embodiment 3C- reactive protein detection kit
Reagent R1:
| Acetate buffer | 70mmol/L |
| Potassium chloride | 15g/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 | 1g/L |
| Goat-anti Human C-reactiveprotein antibody | 250mg/L |
| Nonidet P40 | 5g/L |
| Magnesium sulfate | 0.05g/L |
| Calcium acetate | 0.05g/L |
Calibration object:
C reactive protein antigen standard dilutions (80mmol/L TRIS buffer, 20g/L sodium chloride, 1g/L nitrine
Sodium, 55g/L glucan, 50g/L trehalose, 60g/L sucrose, 70g/L bovine serum albumin(BSA)) dissolution, it is examined with commercially available contrast agents
It surveys 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
C reactive protein standard items (C reactive protein antigen concentration: 2mg/L, 10mg/L, 40mg/L, 60mg/L, 80mg/L).Then it uses
2~8 DEG C of preservations are placed in 0.65 μm of membrane filtration degerming.
4 Retinal-binding protein detection kit of embodiment
Reagent R1:
| MOPS buffer | 150mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Macrogol 6000 | 60g/L |
| Nonidet P40 | 40g/L |
Reagent R2:
| MOPS buffer | 100mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Goat-anti human retinol-binding protein antibody | 500mg/L |
| Nonidet P40 | 40g/L |
| Magnesium sulfate | 10g/L |
| Calcium acetate | 15g/L |
Calibration object:
RBP ELISA antigen standard dilutions (100mmol/L TRIS buffer, 20g/L sodium chloride, 1g/
L Sodium azide, 10g/L glucan, 100g/L trehalose, 100g/L sucrose, 100g/L bovine serum albumin(BSA)) dissolution, with commercially available right
It detects and is adjusted to 160mg/L according to reagent, packing is stored in -20 DEG C.It is diluted to not using preceding taking-up, and with standard dilutions
With RBP ELISA standard items (the RBP ELISA antigen concentration: 0mg/L, 20mg/L, 40mg/L, 70mg/ of concentration
L,100mg/L).Then with 0.45 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
5 rheumatoid factor of embodiment (RF) detection kit
Reagent R1:
| MOPS buffer | 150mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Macrogol 6000 | 60g/L |
| Nonidet P40 | 30g/L |
Reagent R2:
| MOPS buffer | 100mmol/L |
| Sodium chloride | 30g/L |
| Sodium azide | 1g/L |
| Goat anti-human antibody | 1000mg/L |
| Nonidet P40 | 30g/L |
| Magnesium sulfate | 5g/L |
| Calcium acetate | 10g/L |
Calibration object:
Rabbit-anti goat-anti body with standard dilutions (100mmol/L TRIS buffer, 30g/L sodium chloride, 2g/L Sodium azide,
100g/L glucan, 100g/L trehalose, 100g/L sucrose, 100g/L bovine serum albumin(BSA)) dissolution, it is examined with commercially available contrast agents
It surveys and adjusts to 120IU/mL, packing is stored in -20 DEG C.Various concentration is diluted to using preceding taking-up, and with standard dilutions
Rabbit-anti goat-anti body with standard items (rabbit-anti goat-anti bulk concentration: 0IU/mL, 20IU/mL, 50IU/mL, 80IU/mL, 100IU/mL).
Then with 0.65 μm of membrane filtration degerming, 2~8 DEG C of preservations are placed.
6 apolipoprotein E detection kit of embodiment
Reagent R1:
Reagent R2:
| TRIS buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein E antibody | 10mg/L |
| Nonidet P40 | 1g/L |
| Magnesium sulfate | 0.01g/L |
Calibration object:
ApoE antigen with standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L Sodium azide,
0.3g/L glucan, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin(BSA)) dissolution, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.It is marked using preceding taking-up, and with the ApoE that standard dilutions are diluted to various concentration
Quasi- product (ApoE antigen concentration: 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 E detection kit of embodiment
Reagent R1:
| TRIS buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Nonidet P40 | 1g/L |
Reagent R2:
Calibration object:
ApoE antigen with standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L Sodium azide,
0.3g/L glucan, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin(BSA)) dissolution, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.It is marked using preceding taking-up, and with the ApoE that standard dilutions are diluted to various concentration
Quasi- product (ApoE antigen concentration: 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 E detection kit of embodiment
Reagent R1:
| TRIS buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Macrogol 6000 | 1g/L |
| Polysorbas20 | 1g/L |
Reagent R2:
| TRIS buffer | 20mmol/L |
| Sodium chloride | 1g/L |
| Sodium azide | 0.5g/L |
| Goat-anti human apolipoprotein E antibody | 10mg/L |
| Polysorbas20 | 1g/L |
Calibration object:
ApoE antigen with standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L Sodium azide,
0.3g/L glucan, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin(BSA)) dissolution, is detected and is adjusted with commercially available contrast agents
It is whole to 120mg/L, packing is stored in -20 DEG C.It is marked using preceding taking-up, and with the ApoE that standard dilutions are diluted to various concentration
Quasi- product (ApoE antigen concentration: 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 value=STDEV (1-7)/mean value.
1. preparing the sample that a apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 1
7 times, testing result is as shown in table 1.
1:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
Seen from table 1, the apo E concentration that embodiment 1 is measured 1 month, 3 months, 12 months close to true value,
And the coefficient of variation (CV value) measured) it is respectively less than 2%, illustrate that kit of the invention is reproducible, performance is stablized, measurement is quasi-
Really.
2. preparing the sample that a Apolipoprotein A1 concentration is 1mg/L, sample is repeated to detect with the kit of embodiment 2
7 times, testing result is as shown in table 2.
2:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
As can be seen from Table 2, embodiment 2 is close true in the Apolipoprotein A1 concentration that 1 month, 3 months, 12 months measure
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 C reactive protein concentration 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.
3:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
Seen from table 3, the C reactive protein concentration that embodiment 3 was measured 1 month, 3 months, 12 months is close 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. the sample that a RBP ELISA concentration is 70mg/L is prepared, with the kit of embodiment 4 to sample weight
Reinspection is surveyed 7 times, and testing result is as shown in table 4.
4:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
By table 4 as it can be seen that embodiment 4 is close true in the RBP ELISA concentration that 1 month, 3 months, 12 months measure
Real 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 quasi-
Really.
5. preparing the sample that a rheumatoid factor concentration is 25IU/mL, sample is repeated to examine with the kit of embodiment 5
It surveys 7 times, testing result is as shown in table 5.
5:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
By table 5 as it can be seen that embodiment 5 in the rheumatoid factor concentration that 1 month, 3 months, 12 months measure is true value,
And the coefficient of variation measured is respectively less than 0.05%, illustrates that kit of the invention is reproducible, performance is stablized, measurement is accurate.
6. preparing the sample that a apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 6
7 times, testing result is as shown in table 6.
6:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
By table 6 as it can be seen that embodiment 6 deviates true value in the apo E concentration that 1 month, 3 months, 12 months measure,
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 apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 7
7 times, testing result is as shown in table 7.
7:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
By table 7 as it can be seen that embodiment 7 deviates true value in the apo E concentration that 1 month, 3 months, 12 months measure,
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 apo E concentration is 40mg/L, sample is repeated to detect with the kit of embodiment 8
7 times, testing result is as shown in table 8.
8:2-8 DEG C of table preservation 1 month, 3 months, 12 months measurement results
By table 8 as it can be seen that embodiment 8 deviates true value in the apo E concentration that 1 month, 3 months, 12 months measure,
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 Immunoturbidimetric kit, it is characterised in that: the kit includes in reagent R1 and reagent R2, the reagent R1
It include Nonidet P40 1-50g/L, magnesium sulfate in the reagent R2 comprising Nonidet P40 1-50g/L
0.01-15g/L, calcium acetate 0.01-20g/L and antibody 10-1000mg/L；It wherein also include buffer, nothing in the reagent R1
Machine salt, preservative and aggregation also include buffer, inorganic salts and preservative in the reagent R2；And wherein
The antibody is animal anti-human antibody,
The aggregation is one of polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or PEG 8000
Or it is several,
Inorganic salts described in reagent R2 are one or both of sodium chloride, potassium chloride.
2. Immunoturbidimetric kit according to claim 1, it is characterised in that: the Nonidet P40 is 5-
40g/L。
3. Immunoturbidimetric kit according to claim 2, it is characterised in that: the Nonidet P40 is 30g/
L。
4. Immunoturbidimetric kit according to claim 1, it is characterised in that: the magnesium sulfate is 0.05-10g/L.
5. Immunoturbidimetric kit according to claim 4, it is characterised in that: the magnesium sulfate is 5g/L.
6. Immunoturbidimetric kit according to claim 1, it is characterised in that: the calcium acetate is 0.05-15g/L.
7. Immunoturbidimetric kit according to claim 6, it is characterised in that: the calcium acetate is 10g/L.
8. Immunoturbidimetric kit according to claim 1-7, it is characterised in that: also include in the reagent R1
Buffer 20-150mmol/L, inorganic salts 1-30g/L, preservative 0.5-1g/L and aggregation 1-60g/L.
9. Immunoturbidimetric kit according to claim 1-7, it is characterised in that: also include in the reagent R2
Buffer 20-100mmol/L, inorganic salts 1-30g/L and preservative 0.5-1g/L, wherein inorganic salts described in reagent R2 are chlorination
One or both of sodium, potassium chloride.
10. Immunoturbidimetric kit according to claim 1, it is characterised in that: the kit includes calibration object, described
Calibration object is calibrated using multiple spot；The calibration object includes 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.
11. Immunoturbidimetric kit according to claim 1, 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 be acetate buffer, ammonium chloride buffer, phosphate buffer, TRIS buffer, borate buffer,
One or more of glycine buffer, CAPSO, MOPS or Hepes buffer；
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 of polyethylene glycol 2000, Macrogol 4000, Macrogol 6000 or PEG 8000
Or it is several.
12. Immunoturbidimetric kit according to claim 1, it is characterised in that: the kit carries rouge egg for detecting
White A1, apolipoprotein B, immunoglobulin A, immunoglobulin G, immunoglobulin M, Complement C_3, Complement C4, apo E, C-
Reactive protein, RBP ELISA, rheumatoid factor, immunoglobulin E, prealbumin or lipoprotein a.
13. a kind of detection method using Immunoturbidimetric kit described in claim 1, includes the following steps:
(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:(1-6 by volume) it 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 test substance in sample to be tested automatically.
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| CN109856067A (en) * | 2019-01-11 | 2019-06-07 | 河北省医疗器械与药品包装材料检验研究院(河北省医疗器械技术审评中心) | A kind of c reactive protein assay kit |
| CN111693699A (en) * | 2020-07-07 | 2020-09-22 | 上海怡珏生物科技有限公司 | Application of C4 antibody in preparation of detection kit |
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