CN107490675A - A kind of Immunoturbidimetric kit and detection method - Google Patents

Abstract

The present invention provides a kind of Immunoturbidimetric kit, it includes reagent R1 and reagent R2, included in wherein reagent R1 in Nonidet P40 1 50g/L, reagent R2 and include the 50g/L of Nonidet P40 1, the 15g/L of magnesium salts 0.01, the 20g/L of calcium acetate 0.01 and antibody.Kit antibody performance of the invention is good, reproducible, reagent testing result is accurate, disclosure satisfy that requirement.

Description

Immunity turbidimetry kit and detection method

Technical Field

The invention relates to the field of medical immune in-vitro diagnosis, in particular to an immunoturbidimetric kit and a detection method.

Background

Turbidimetry is widely used in clinical testing. Immunoturbidimetry is now the most commonly used.

Most of the early immunoassay techniques analyze the presence or absence and content of specific proteins in a sample to be detected by observing the formation of precipitates, agglutination and hemolysis and measuring light scattering caused by aggregates, such as immunodiffusion, immunoelectrophoresis, direct and brief hemagglutination, passive hemagglutination, complement fixation experiments, etc., and these detection methods have the advantages of low cost, easy judgment of results, and easy technical mastery, and can be widely used for detecting various types of clinical samples. However, the above-mentioned methods tend to be eliminated due to their cumbersome operation, time-consuming and poor sensitivity and accuracy.

Immunoturbidimetry overcomes the above disadvantages and allows the development of automated instruments with precise quantitation in combination with clinical requirements. Therefore, for immunological detection, the immunoturbidimetry has the specificity of combining immunological antigen and antibody, has the characteristic of biochemical reaction, can detect a trace amount of substance to be detected in body fluid, particularly blood, on an automatic biochemical instrument, and is a practical clinical test technology with application prospect.

Immunoturbidimetry (Turbidimetric inhibition assay) is an antigen-antibody binding kinetic assay. The basic principle is as follows: when the antigen and antibody react in a special dilution system and in a suitable ratio (generally, an excess amount of antibody is specified), the formed soluble immune complex precipitates from the liquid phase under the action of the aggregation promoter in the dilution system to form microparticles, and turbidity appears in the reaction solution. When the antibody concentration is fixed, the amount of the immunocomplex formed increases with the increase in the amount of the antigen in the sample, and the turbidity of the reaction solution also increases. The content of the antigen in the sample can be calculated by measuring the turbidity of the reaction solution and comparing with a series of standard products.

Various detection instruments developed and developed based on the basic principle of immunoturbidimetry have been widely used in many aspects of clinical examination, which is the basic working principle of optical methods and immunological methods of blood coagulators; the method is a determination principle of apolipoprotein, hapten and other proteins in full-automatic biochemical determination; meanwhile, the method can also be applied to microorganism detection. By accurately quantifying a variety of substances, there is great clinical significance in the diagnosis, treatment and prognosis evaluation of many diseases.

The clinical commonly used immunoturbidimetry can directly analyze samples in batches on a full-automatic biochemical analyzer due to small sample consumption, and is simple to operate, but the currently established reagents and methods have some defects and shortcomings, which are mainly shown in that:

the antibody is easy to generate flocculent or flaky precipitation in the preservation process, so that the performance of the antibody is poor, the repeatability is poor, and the Coefficient of Variation (CV) is increased, the detection result of the reagent is inaccurate, a filtering step is added, the operation is complex, the antibody can be removed by filtering, the detection effect is influenced, a certain influence is caused on a patient, and the use requirement cannot be met.

Disclosure of Invention

In order to solve the problems, the invention discloses an immunoturbidimetric kit and a detection method, and the kit has the advantages of good reagent stability, good uniformity, good detection result accuracy, convenience in operation and convenience in popularization and application.

In order to achieve the purpose, the invention provides the following technical scheme:

the invention provides an immunoturbidimetric kit, which comprises a reagent R1 and a reagent R2,

the reagent R1 contains 1-50g/L of ethyl phenyl polyethylene glycol, preferably 5-40g/L, and more preferably 30 g/L;

the reagent R2 contains 1-50g/L of ethyl phenyl polyethylene glycol, preferably 5-40g/L, and more preferably 30 g/L;

the reagent R2 comprises magnesium salt 0.01-15g/L, preferably 0.05-10g/L, and more preferably 5 g/L; the magnesium salt is preferably one or more of magnesium sulfate, magnesium chloride or magnesium acetate.

The reagent R2 contains 0.01-20g/L of calcium acetate, preferably 0.05-15g/L, and more preferably 10 g/L.

The reagent R2 contains 10-1000mg/L antibody.

Wherein,

the reagent R1 also comprises buffer solution, inorganic salt, preservative and aggregate; preferably, the reagent R1 further comprises buffer solution 20-150mmol/L, inorganic salt 1-30g/L, preservative 0.5-1g/L and aggregate 1-60 g/L;

the reagent R2 also comprises buffer solution, inorganic salt and preservative; preferably, the reagent R2 further comprises a buffer solution 20-100mmol/L, an inorganic salt 1-30g/L and a preservative 0.5-1 g/L.

The kit comprises a calibrator, and the calibrator is calibrated by adopting multiple points; the calibrator comprises 20-100mmol/L of buffer solution, 1-30g/L of inorganic salt, 0.1-2g/L of preservative, 0.3-100g/L of glucan, 1-100g/L of trehalose, 1-100g/L of sucrose, 1-100g/L of bovine serum albumin and antigen.

Preferably, the antibody is a goat anti-human, rabbit anti-human, horse anti-human, mouse anti-human or other animal anti-human antibody.

Preferably, the buffer solution is one or more of acetate buffer solution, ammonium chloride buffer solution, phosphate buffer solution, TRIS buffer solution, boric acid buffer solution, glycine buffer solution, CAPSO, MOPS or Hepes buffer solution.

Preferably, the inorganic salt is one or both of sodium chloride and potassium chloride.

Preferably, the preservative is one or more of sodium azide, phenol, p-hydroxybenzoic acid, ethyl p-hydroxybenzoate or ethylmercuric thiosulfate.

Preferably, the aggregate is one or more of polyethylene glycol 2000, polyethylene glycol 4000, polyethylene glycol 6000 or polyethylene glycol 8000.

Wherein the kit is used for detecting apolipoprotein A1, apolipoprotein B, immunoglobulin A, immunoglobulin G, immunoglobulin M, complement C3, complement C4, apolipoprotein E, C-reactive protein, retinol binding protein, rheumatoid factor, immunoglobulin E, prealbumin or lipoprotein a.

The invention also provides a detection method using the immunoturbidimetric kit, which comprises the following steps:

(1) adding the reagent R1 into a sample to be detected, and uniformly mixing, wherein the volume ratio of the sample to be detected to the reagent R1 is (1-9): 300, incubating at 37 ℃, and reading the absorbance A1 at a certain wavelength;

(2) adding a reagent R2 into the mixed solution obtained in the step (1), and uniformly mixing, wherein the volume ratio of the reagent R2 to the reagent R1 is 1: (1-6) adding, incubating at 37 ℃, and reading the absorbance A2 at a certain wavelength;

(3) obtaining absorbance delta A, wherein the absorbance delta A is A2-A1;

(4) and (3) fitting a standard curve on a full-automatic biochemical analyzer by using the calibrator through a built-in curve fitting mode, and automatically calculating the content of the substance to be detected in the sample to be detected according to the absorbance.

The aggregate referred to in the present invention is a substance that promotes aggregation of antigen and antibody in a reaction.

The immunoturbidimetric kit and the detection method provided by the invention relate to the following raw material sources:

due to the adoption of the technical scheme, the invention has the beneficial effects that:

the immunoturbidimetric kit and the detection method provided by the invention have the advantages of good reagent stability, good uniformity, stable storage of antibodies, no precipitation phenomenon, high antibody titer, good performance, no filtration step, convenience in operation, low cost, accurate detection result, good repeatability and wider universality.

Drawings

Figure 1 shows the 1 month storage results of example 5.

Figure 2 shows the results of example 5 stored for 3 months.

Figure 3 shows the 12 month storage results of example 5.

Figure 4 shows the 1 month storage results of example 8.

FIG. 5 shows the results of example 8 with 3 months of storage

Figure 6 shows the 12 month results of example 8.

Detailed description of the preferred embodiments

In order to make the technical solutions in the present application better understood, the present invention is further described below with reference to examples, and it is obvious that the described examples are only a part of the examples of the present application, but not all of the examples. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

Example 1 detection kit for Apolipoprotein E

Reagent R1:

TRIS buffer	20mmol/L
		Sodium chloride	1g/L
Sodium azide	0.5g/L
		Polyethylene glycol 6000	1g/L
Ethyl phenyl polyethylene glycol	1g/L

Reagent R2:

TRIS buffer	20mmol/L
		Sodium chloride	1g/L
Sodium azide	0.5g/L
		Sheep anti-human apolipoprotein E antibody	10mg/L
Ethyl phenyl polyethylene glycol	1g/L
		Magnesium sulfate	0.01g/L
Calcium acetate	0.01g/L

Calibration products:

ApoE antigen was dissolved in standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L sodium azide, 0.3g/L dextran, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin), assayed with commercially available control reagents and adjusted to 120mg/L, stored at-20 ℃. Before use, the samples were taken out and diluted with standard dilutions to ApoE standards of different concentrations (ApoE antigen concentration: 0mg/L, 10mg/L, 20mg/L, 50mg/L, 70 mg/L). Then filtering and sterilizing by using a filter membrane with the diameter of 0.65 mu m, and storing at the temperature of 2-8 ℃.

Example 2 detection kit for apolipoprotein A1

Reagent R1:

phosphate buffer	30mmol/L
		Potassium chloride	5g/L
Phenol and its preparation	0.7g/L
		Polyethylene glycol 6000	20g/L
Ethyl phenyl polyethylene glycol	50g/L

Reagent R2:

phosphate buffer	40mmol/L
		Potassium chloride	5g/L
Phenol and its preparation	0.7g/L
		Sheep anti-human apolipoprotein A1 antibody	50mg/L
Ethyl phenyl polyethylene glycol	50g/L
		Magnesium sulfate	15g/L
Calcium acetate	20g/L

Calibration products:

the apolipoprotein A1 antigen is dissolved by standard diluent (40mmol/L glycine buffer solution, 10g/L sodium chloride, 0.7g/L sodium azide, 20g/L dextran, 25g/L trehalose, 30g/L sucrose, 20g/L bovine serum albumin), detected by a commercial control reagent, adjusted to 100mg/L, and stored at-20 ℃. Before use, the samples were taken out and diluted with standard dilutions to different concentrations of apolipoprotein A1 standard (apolipoprotein A1 antigen concentration: 0mg/L, 10mg/L, 30mg/L, 50mg/L, 70 mg/L). Then filtering and sterilizing by using a filter membrane with the diameter of 0.65 mu m, and storing at the temperature of 2-8 ℃.

Example 3C-reactive protein detection kit

Reagent R1:

acetate buffer solution	70mmol/L
		Potassium chloride	15g/L
Sodium azide	0.8g/L
		Polyethylene glycol 6000	50g/L
Ethyl phenyl polyethylene glycol	5g/L

Reagent R2:

acetate buffer solution	80mmol/L
		Potassium chloride	20g/L
Sodium azide	1g/L
		Anti-human C-reactive protein antibody for sheep	250mg/L
Ethyl phenyl polyethylene glycol	5g/L
		Magnesium sulfate	0.05g/L
Calcium acetate	0.05g/L

Calibration products:

the C-reactive protein antigen was dissolved in standard dilutions (80mmol/L TRIS buffer, 20g/L sodium chloride, 1g/L sodium azide, 55g/L dextran, 50g/L trehalose, 60g/L sucrose, 70g/L bovine serum albumin), assayed with commercially available control reagents and adjusted to 200mg/L, stored at-20 ℃. Before use, the sample was taken out and diluted with a standard dilution to obtain C-reactive protein standards (C-reactive protein antigen concentration: 2mg/L, 10mg/L, 40mg/L, 60mg/L, 80mg/L) of different concentrations. Then filtering and sterilizing by using a filter membrane with the diameter of 0.65 mu m, and storing at the temperature of 2-8 ℃.

Example 4 Retinol binding protein assay kit

Reagent R1:

MOPS buffer solution	150mmol/L
		Sodium chloride	30g/L
Sodium azide	1g/L
		Polyethylene glycol 6000	60g/L
Ethyl phenyl polyethylene glycol	40g/L

Reagent R2:

MOPS buffer solution	100mmol/L
		Sodium chloride	30g/L
Sodium azide	1g/L
		Goat anti-human retinol binding protein antibody	500mg/L
Ethyl phenyl polyethylene glycol	40g/L
		Magnesium sulfate	10g/L
Calcium acetate	15g/L

Calibration products:

retinol binding protein antigen was dissolved in standard diluent (100mmol/L TRIS buffer, 20g/L sodium chloride, 1g/L sodium azide, 10g/L dextran, 100g/L trehalose, 100g/L sucrose, 100g/L bovine serum albumin), detected with a commercially available control reagent, adjusted to 160mg/L, stored at-20 ℃. Before use, the samples were taken out and diluted with standard dilution to different concentrations of retinol binding protein standard (retinol binding protein antigen concentrations: 0mg/L, 20mg/L, 40mg/L, 70mg/L, 100 mg/L). Then filtering and sterilizing by using a filter membrane with the diameter of 0.45 mu m, and storing at the temperature of 2-8 ℃.

Example 5 detection kit for Rheumatoid Factor (RF)

Reagent R1:

MOPS buffer solution	150mmol/L
		Sodium chloride	30g/L
Sodium azide	1g/L
		Polyethylene glycol 6000	60g/L
Ethyl phenyl polyethylene glycol	30g/L

Reagent R2:

MOPS buffer solution	100mmol/L
		Sodium chloride	30g/L
Sodium azide	1g/L
		Goat anti-human antibodies	1000mg/L
Ethyl phenyl polyethylene glycol	30g/L
		Magnesium sulfate	5g/L
Calcium acetate	10g/L

Calibration products:

dissolving rabbit anti-goat antibody with standard diluent (100mmol/L TRIS buffer solution, 30g/L sodium chloride, 2g/L sodium azide, 100g/L dextran, 100g/L trehalose, 100g/L sucrose, 100g/L bovine serum albumin), detecting with commercial control reagent, adjusting to 120IU/mL, subpackaging and storing at-20 deg.C. Before use, the antibody is taken out and diluted into standard substances for rabbit anti-sheep antibodies with different concentrations by using a standard substance diluent (the concentration of the rabbit anti-sheep antibodies is 0IU/mL, 20IU/mL, 50IU/mL, 80IU/mL and 100 IU/mL). Then filtering and sterilizing by using a filter membrane with the diameter of 0.65 mu m, and storing at the temperature of 2-8 ℃.

Example 6 detection kit for Apolipoprotein E

Reagent R1:

reagent R2:

TRIS buffer	20mmol/L
		Sodium chloride	1g/L
Sodium azide	0.5g/L
		Sheep anti-human apolipoprotein E antibody	10mg/L
Ethyl phenyl polyethylene glycol	1g/L
		Magnesium sulfate	0.01g/L

Calibration products:

ApoE antigen was dissolved in standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L sodium azide, 0.3g/L dextran, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin), assayed with commercially available control reagents and adjusted to 120mg/L, stored at-20 ℃. Before use, the samples were taken out and diluted with standard dilutions to ApoE standards of different concentrations (ApoE antigen concentration: 0mg/L, 10mg/L, 20mg/L, 50mg/L, 70 mg/L). Then filtering and sterilizing by using a filter membrane with the diameter of 0.65 mu m, and storing at the temperature of 2-8 ℃.

Example 7 Apolipoprotein E detection kit

Reagent R1:

TRIS buffer	20mmol/L
		Sodium chloride	1g/L
Sodium azide	0.5g/L
		Polyethylene glycol 6000	1g/L
Ethyl phenyl polyethylene glycol	1g/L

Reagent R2:

calibration products:

ApoE antigen was dissolved in standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L sodium azide, 0.3g/L dextran, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin), assayed with commercially available control reagents and adjusted to 120mg/L, stored at-20 ℃. Before use, the samples were taken out and diluted with standard dilutions to ApoE standards of different concentrations (ApoE antigen concentration: 0mg/L, 10mg/L, 20mg/L, 50mg/L, 70 mg/L). Then filtering and sterilizing by using a filter membrane with the diameter of 0.65 mu m, and storing at the temperature of 2-8 ℃.

Example 8 detection kit for Apolipoprotein E

Reagent R1:

TRIS buffer	20mmol/L
		Sodium chloride	1g/L
Sodium azide	0.5g/L
		Polyethylene glycol 6000	1g/L
Tween 20	1g/L

Reagent R2:

TRIS buffer	20mmol/L
		Sodium chloride	1g/L
Sodium azide	0.5g/L
		Sheep anti-human apolipoprotein E antibody	10mg/L
Tween 20	1g/L

Calibration products:

ApoE antigen was dissolved in standard dilutions (20mmol/L phosphate buffer, 1g/L sodium chloride, 0.1g/L sodium azide, 0.3g/L dextran, 1g/L trehalose, 1g/L sucrose, 1g/L bovine serum albumin), assayed with commercially available control reagents and adjusted to 120mg/L, stored at-20 ℃. Before use, the samples were taken out and diluted with standard dilutions to ApoE standards of different concentrations (ApoE antigen concentration: 0mg/L, 10mg/L, 20mg/L, 50mg/L, 70 mg/L). Then filtering and sterilizing by using a filter membrane with the diameter of 0.65 mu m, and storing at the temperature of 2-8 ℃. Comparing the detection results of different embodiments, wherein the CV value is STDEV (1-7)/mean value.

1. A sample with an apolipoprotein E concentration of 40mg/L was prepared, and the test of the sample was repeated 7 times using the kit of example 1, with the test results shown in Table 1.

Table 1: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 1, the apolipoprotein E concentrations measured in example 1 in 1 month, 3 months and 12 months are all close to the true values, and the measured variation coefficients (CV values)) are all less than 2%, which indicates that the kit provided by the invention has good repeatability, stable performance and accurate measurement.

2. A sample with an apolipoprotein A1 concentration of 1mg/L was prepared, and the assay was repeated 7 times using the kit of example 2, with the assay results shown in Table 2.

Table 2: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 2, the apolipoprotein A1 concentrations measured in 1 month, 3 months and 12 months in the example 2 are close to the true values, and the measured variation coefficients are less than 2%, which indicates that the kit has good repeatability, stable performance and accurate measurement.

3. A sample with a C-reactive protein concentration of 25mg/L was prepared, and the assay was repeated 7 times using the kit of example 3, with the assay results shown in Table 3.

Table 3: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 3, the concentrations of C-reactive protein measured in 1 month, 3 months and 12 months in example 3 are all close to the true values, and the measured variation coefficients are all less than 1%, which indicates that the kit of the invention has good repeatability, stable performance and accurate measurement.

4. A sample with a retinol binding protein concentration of 70mg/L was prepared, and the assay was repeated 7 times using the kit of example 4, with the assay results shown in Table 4.

Table 4: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 4, the retinol binding protein concentrations measured in 1 month, 3 months and 12 months in example 4 are all close to the true values, and the measured variation coefficients are all less than 1%, which indicates that the kit of the invention has good repeatability, stable performance and accurate measurement.

5. A sample with rheumatoid factor concentration of 25IU/mL is prepared, and the detection is repeated for 7 times by using the kit of example 5, and the detection results are shown in Table 5.

Table 5: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 5, the concentrations of the rheumatoid factors measured in 1 month, 3 months and 12 months in example 5 are all true values, and the measured variation coefficients are all less than 0.05%, which indicates that the kit of the invention has good repeatability, stable performance and accurate measurement.

6. A sample with an apolipoprotein E concentration of 40mg/L was prepared, and the test of the sample was repeated 7 times using the kit of example 6, with the test results shown in Table 6.

Table 6: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 6, the apolipoprotein E concentrations measured in example 6 in 1 month, 3 months and 12 months all deviate from the true values, and the measured variation coefficients are all more than 4%, which indicates that the kit has poor repeatability, unstable performance and inaccurate measurement.

7. A sample with an apolipoprotein E concentration of 40mg/L was prepared, and the sample was repeatedly tested 7 times using the kit of example 7, with the test results shown in Table 7.

Table 7: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 7, the apolipoprotein E concentrations measured in example 7 in 1 month, 3 months and 12 months all deviate from the true values, and the measured variation coefficients are all more than 4%, which indicates that the kit has poor repeatability, unstable performance and inaccurate measurement.

8. A sample with an apolipoprotein E concentration of 40mg/L was prepared, and the test of the sample was repeated 7 times using the kit of example 8, with the test results shown in Table 8.

Table 8: the measurement result of the storage at 2-8 ℃ for 1 month, 3 months and 12 months

As can be seen from Table 8, the apolipoprotein E concentrations measured in example 8 in 1 month, 3 months and 12 months all deviate from the true values, and the measured variation coefficients are all more than 5%, which indicates that the kit has poor repeatability, unstable performance and inaccurate measurement.

It is to be understood that the invention disclosed is not limited to the particular methodology, protocols, and materials described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

Those skilled in the art will also recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims (10)

1. An immunoturbidimetric kit, which is characterized in that: the kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises 1-50g/L of ethylphenyl polyethylene glycol, and the reagent R2 comprises 1-50g/L of ethylphenyl polyethylene glycol, 0.01-15g/L of magnesium salt, 0.01-20g/L of calcium acetate and 10-1000mg/L of antibody.

2. The immunoturbidimetric kit according to claim 1, characterized in that: the ethyl phenyl polyethylene glycol is 5-40g/L, and preferably 30 g/L.

3. The immunoturbidimetric kit according to claim 1 or 2, characterized in that: the magnesium salt is 0.05-10g/L, preferably 5 g/L; the magnesium salt is preferably one or more of magnesium sulfate, magnesium chloride or magnesium acetate.

4. The immunoturbidimetric kit according to any one of claims 1 to 3, characterized in that: the calcium acetate is 0.05-15g/L, preferably 10 g/L.

5. The immunoturbidimetric kit according to any one of claims 1 to 4, characterized in that: the reagent R1 also comprises buffer solution, inorganic salt, preservative and aggregate; preferably, the reagent R1 further comprises buffer solution 20-150mmol/L, inorganic salt 1-30g/L, preservative 0.5-1g/L and aggregate 1-60 g/L.

6. The immunoturbidimetric kit according to any one of claims 1 to 5, characterized in that: the reagent R2 further comprises a buffer solution, an inorganic salt and a preservative, and preferably, the reagent R2 further comprises 20-100mmol/L of the buffer solution, 1-30g/L of the inorganic salt and 0.5-1g/L of the preservative.

7. The immunoturbidimetric kit according to any one of claims 1 to 6, wherein: the kit comprises a calibrator, and the calibrator is calibrated by adopting multiple points; the calibrator comprises 20-100mmol/L of buffer solution, 1-30g/L of inorganic salt, 0.1-2g/L of preservative, 0.3-100g/L of glucan, 1-100g/L of trehalose, 1-100g/L of sucrose, 1-100g/L of bovine serum albumin and antigen.

8. The immunoturbidimetric kit according to any one of claims 1 to 7, characterized in that:

the antibody is preferably a sheep anti-human antibody, a rabbit anti-human antibody, a horse anti-human antibody, a mouse anti-human antibody or other animal anti-human antibody;

the buffer solution is preferably one or more of acetate buffer solution, ammonium chloride buffer solution, phosphate buffer solution, TRIS buffer solution, boric acid buffer solution, glycine buffer solution, CAPSO, MOPS or Hepes buffer solution; the inorganic salt is preferably one or two of sodium chloride and potassium chloride;

the preservative is preferably one or more of sodium azide, phenol, p-hydroxybenzoic acid, ethyl p-hydroxybenzoate or ethylmercuric thiosulfate;

the aggregate is preferably one or more of polyethylene glycol 2000, polyethylene glycol 4000, polyethylene glycol 6000 or polyethylene glycol 8000.

9. The immunoturbidimetric kit according to any one of claims 1 to 8, characterized in that: the kit is used for detecting apolipoprotein A1, apolipoprotein B, immunoglobulin A, immunoglobulin G, immunoglobulin M, complement C3, complement C4, apolipoprotein E, C-reactive protein, retinol binding protein, rheumatoid factor, immunoglobulin E, prealbumin or lipoprotein a.

10. A detection method using the immunoturbidimetric kit comprises the following steps:

(1) adding the reagent R1 into a sample to be detected, and uniformly mixing, wherein the volume ratio of the sample to be detected to the reagent R1 is (1-9): 300, incubating at 37 ℃, and reading the absorbance A1 at a certain wavelength;

(2) adding a reagent R2 into the mixed solution obtained in the step (1), and uniformly mixing, wherein the volume ratio of the reagent R2 to the reagent R1 is 1: (1-6) adding, incubating at 37 ℃, and reading the absorbance A2 at a certain wavelength;

(3) obtaining absorbance delta A, wherein the absorbance delta A is A2-A1;

(4) and (3) fitting a standard curve on a full-automatic biochemical analyzer by using the calibrator through a built-in curve fitting mode, and automatically calculating the content of the substance to be detected in the sample to be detected according to the absorbance.