CN116519946A - C-reactive protein assay kit with stable bottle opening performance and detection method - Google Patents

Abstract

The invention relates to the technical field of immunonephelometry in-vitro diagnosis, in particular to a C-reactive protein assay kit with stable bottle opening performance and a detection method.

Description

C-reactive protein assay kit with stable bottle opening performance and detection method

Technical Field

The invention relates to the technical field of immunonephelometry in-vitro diagnosis, in particular to a C-reactive protein assay kit with stable bottle opening performance and a detection method.

Background

In clinic, an index in blood detection is C-reactive protein, CRP for short, and cardiovascular related conditions are hypertension patients and diabetes patients in general, and whether inflammation occurs in vivo or not, the degree of elevation and the severity of the inflammation can be predicted by a C-reactive protein test. If the C-reactive protein is obviously increased, vascular inflammation may exist, atherosclerosis is easy to occur, coronary heart disease and cerebrovascular disease are easy to occur, most of early immunoassay techniques analyze the presence and the content of specific proteins in a sample to be detected through the formation of precipitates, the occurrence of agglutination and hemolysis phenomena and determination of light scattering caused by aggregates, but the methods have complicated operation and poor accuracy.

The immunoturbidimetry is an antigen-antibody binding dynamic assay method which is based on the principle that: when the antigen and the antibody react in a special dilution system and the proportion is proper, the formed soluble immune complex is separated out from a liquid phase under the action of a polymerization promoter in the dilution system to form particles, and the reaction liquid is turbid. When the concentration of the antibody is fixed, the amount of the immunocomplex formed increases with an increase in the amount of the antigen in the sample, and the turbidity of the reaction solution increases.

Most of the kits on the market at present, such as a latex-enhanced turbidimetric immunoassay reagent, a kit and application thereof, have the patent number of CN 201310079651.X; the second one is a kit for measuring latex immune turbidimetry and its application, the patent number is CN 202010528691.8, the third one is a kit for enhancing latex sensitivity CRP, the kit is CN 202110864091.3, the kit base is sensitization latex immune turbidimetry, and the antibodies are easy to precipitate in the preservation process, so that the antibody performance is poor, the repeatability is poor, the variation coefficient is large, and the reagent detection result is inaccurate.

Disclosure of Invention

The invention aims to provide a C-reactive protein assay kit with stable bottle opening performance and a detection method, and aims to solve the problem that the existing immunonephelometry assay kit is poor in detection precision.

To achieve the above object, in a first aspect, the present invention provides a C-reactive protein assay kit with stable performance in opening bottles, comprising a first reagent, a second reagent and a standard diluent for complement CRP antigen.

Wherein the first reagent, the second reagent and the standard substance diluent for complement CRP antigen are all preserved at the temperature of 2-8 ℃.

In a second aspect, the invention also provides a method for detecting the C-reactive protein assay kit with stable bottle opening performance, which comprises the following steps:

adding the first reagent into a sample to be detected, uniformly mixing, incubating at 37 ℃ to obtain a first mixed solution, and reading absorbance A1 of the first mixed solution at a wavelength of 340 nm;

adding the second reagent into the first mixed solution, uniformly mixing, incubating at 37 ℃ to obtain a second mixed solution, and reading absorbance A2 of the second mixed solution at a wavelength of 340 nm;

obtaining the difference value between the absorbance A2 and the absorbance A1 to obtain an absorbance difference;

fitting a standard curve on a full-automatic biochemical analyzer by using the standard substance diluent for the complement CRP antigen, and calculating the content of complement CRP in the sample to be detected based on the absorbance difference.

Wherein, the volume ratio of the sample to be tested to the first reagent is 17:300, the volume ratio of the second reagent to the first reagent is 1:4.

in a third aspect, the invention also provides a preparation method of the C-reactive protein assay kit with stable bottle opening performance, which comprises the following steps:

obtaining polyethylene glycol, sodium azide, tween, purified water, disodium hydrogen phosphate and sodium dihydrogen phosphate, uniformly mixing at 25+/-3 ℃ and filtering to obtain a first reagent;

obtaining water-soluble sodium channel, triton, trehalose, glycerol, sodium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium azide, citric acid, CRP sheep anti-human antibody and purified water, uniformly mixing at 25+/-3 ℃ and filtering to obtain a second reagent;

obtaining citric acid monohydrate, sodium chloride, sodium azide, sodium hydroxide, triton, disodium hydrogen phosphate, sodium dihydrogen phosphate, polyvinylpyrrolidone, sodium complexing agent and purified water, and diluting the citric acid monohydrate, sodium chloride, sodium azide, sodium hydroxide, triton, sodium complexing agent and purified water into complement CRP calibrator with different concentrations by using a calibrator to obtain a standard diluent for complement CRP antigen;

and packaging the first reagent, the second reagent and the complement CRP antigen with a standard substance diluent to obtain a kit.

According to the C-reactive protein determination kit and the detection method, the sample to be detected is mixed with the first reagent to obtain absorbance, the mixed solution is mixed with the second reagent again to obtain absorbance, the difference of the absorbance is obtained again, and finally the standard curve is fitted on a full-automatic biochemical instrument through a built-in curve fitting mode by utilizing the standard substance diluent for the complement CRP antigen, so that the content of the complement CRP in the sample to be detected is calculated according to the absorbance difference.

Drawings

In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic diagram of a C-reactive protein assay kit with stable bottle opening performance.

FIG. 2 is a flow chart of a method for detecting a C-reactive protein assay kit with stable bottle opening performance.

FIG. 3 is a flow chart of a method for preparing a C-reactive protein assay kit with stable opening performance.

In the figure: 1-first reagent, 2-second reagent, 3-complement CRP antigen with standard dilution.

Detailed Description

Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative and intended to explain the present invention and should not be construed as limiting the invention.

Referring to fig. 1, in a first aspect, the present invention provides a C-reactive protein assay kit with stable performance in opening bottles, which comprises a first reagent 1, a second reagent 2 and a standard diluent 3 for complement CRP antigen.

In this embodiment, the first reagent 1, the second reagent 2 and the standard substance dilution 3 for complement CRP antigen are all stored at 2-8 ℃, the standard substance dilution 3 for complement CRP antigen (s1=0 mg/L; s1=9.8 mg/L; s2=28.1 mg/L; s3=56.2 mg/L; s1=115.2 mg/L) is obtained by mixing the sample to be tested with the first reagent 1, the mixed solution is mixed again with the second reagent 2 to obtain absorbance again, and the difference of the absorbance is obtained for two times, finally, the standard curve is fitted on a fully automatic biochemical analyzer by using the standard substance dilution 3 for complement CRP antigen, and the content of complement CRP in the sample to be tested is calculated according to the difference of absorbance.

Referring to fig. 2, in a second aspect, the present invention further provides a method for detecting a C-reactive protein assay kit with stable bottle opening performance, comprising the following steps:

s01, adding the first reagent 1 into a sample to be detected, uniformly mixing, incubating at 37 ℃ to obtain a first mixed solution, and reading absorbance A1 of the first mixed solution at 340nm wavelength;

specifically, the volume ratio of the sample to be tested to the first reagent 1 is 17:300.

s02, adding the second reagent 2 into the first mixed solution, uniformly mixing, incubating at 37 ℃ to obtain a second mixed solution, and reading absorbance A2 of the second mixed solution at a wavelength of 340 nm;

specifically, the volume ratio of the second reagent 2 to the first reagent 1 is 1:4.

s03, obtaining a difference value between the absorbance A2 and the absorbance A1 to obtain an absorbance difference;

specifically, the difference Δa in absorbance, Δa=a2-A1, is obtained.

S04, fitting a standard curve on a full-automatic biochemical analyzer by using the standard substance diluent (3) for the complement CRP antigen, and calculating the content of the complement CRP in the sample to be detected based on the absorbance difference.

Referring to fig. 3, in a third aspect, the present invention further provides a preparation method of a C-reactive protein assay kit with stable bottle opening performance, comprising the following steps:

s11, obtaining polyethylene glycol, sodium azide, tween, purified water, disodium hydrogen phosphate and sodium dihydrogen phosphate, uniformly mixing at 25+/-3 ℃ and filtering to obtain a first reagent 1;

specifically, 5-6 parts of polyethylene glycol, 3-12 parts of sodium azide, 2-15 parts of tween, 1000-1650 parts of purified water, 0.45-0.55 parts of disodium hydrogen phosphate and 15-23 parts of sodium dihydrogen phosphate are respectively weighed, uniformly mixed at 25+/-3 ℃ and filtered to obtain a first reagent 1.

S12, obtaining water-soluble sodium channel, triton, trehalose, glycerol, sodium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium azide, citric acid, CRP sheep anti-human antibody and purified water, uniformly mixing at 25+/-3 ℃ and filtering to obtain a second reagent 2;

specifically, 3-8 parts of water-soluble sodium channel, 1-4 parts of triton, 2-5 parts of trehalose, 2-6 parts of glycerol, 1-4 parts of sodium chloride, 1-3 parts of disodium hydrogen phosphate, 1-2 parts of sodium dihydrogen phosphate, 1-2 parts of sodium azide, 1-3 parts of citric acid, 3-20 parts of CRP sheep anti-human antibody and 40-55 parts of purified water are respectively weighed, uniformly mixed at 25+/-3 ℃ and filtered to obtain a second reagent 2;

s13, obtaining citric acid monohydrate, sodium chloride, sodium azide, sodium hydroxide, triton, disodium hydrogen phosphate, sodium dihydrogen phosphate, polyvinylpyrrolidone, sodium complexing agent and purified water, and diluting the citric acid monohydrate, sodium chloride, sodium azide, sodium hydroxide, triton, sodium complexing agent and purified water into complement CRP calibrator with different concentrations by using a calibrator to obtain a standard product diluent 3 for complement CRP antigen;

specifically, 3 to 6 parts of citric acid monohydrate, 5 to 18 parts of sodium chloride, 1 to 6 parts of sodium azide, 2.3 to 6 parts of sodium hydroxide, 3.7 to 9 parts of triatomic acid, 2 to 15 parts of disodium hydrogen phosphate, 3 to 16 parts of sodium dihydrogen phosphate, 3 to 7 parts of polyvinylpyrrolidone, 3 to 8 parts of sodium complexing agent and 40 to 55 parts of purified water are respectively weighed, and the mixture is diluted into complement CRP calibrator with different concentrations by using a calibrator to obtain a standard diluent 3 for complement CRP antigen;

s14, packaging the first reagent 1, the second reagent 2 and the complement CRP antigen with a standard substance diluent 3 to obtain a kit.

The sources of the raw materials involved in the complement CRP detection kit provided by the invention are as follows:

raw materials	Source
		Polyethylene glycol	Guangdong Guanghua Sci-Tech Co.,Ltd.
Sodium azide	SHANGHAI SEEBIO BIOTECH Inc.
		Tween-type oil	SHANGHAI SEEBIO BIOTECH Inc.
Disodium hydrogen phosphate	SHANGHAI SEEBIO BIOTECH Inc.
		Sodium dihydrogen phosphate	SHANGHAI SEEBIO BIOTECH Inc.
Sodium chloride	SHANGHAI SEEBIO BIOTECH Inc.
		Citric acid	SHANGHAI SEEBIO BIOTECH Inc.
CRP sheep anti-human antibody	Hall (Beijing) diagnostic technology Co., ltd
		Sodium hydroxide	SHANGHAI SEEBIO BIOTECH Inc.
Polyvinyl pyrrolidone	SHANGHAI SEEBIO BIOTECH Inc.
		Cursor-pull medicine	SHANGHAI SEEBIO BIOTECH Inc.
Complex protein sodium	Hall (Beijing) diagnostic technology Co., ltd
		Cursor-pull medicine	SHANGHAI SEEBIO BIOTECH Inc.
Trehalose	SHANGHAI SEEBIO BIOTECH Inc.
		Glycerol	SHANGHAI SEEBIO BIOTECH Inc.

Specific examples are as follows:

example 1

Complement CRP detection kit

First reagent

Reagent name	Dosage of
		Polyethylene glycol	3 parts of
Sodium azide	4 parts of
		Tween-type oil	1 part of
Disodium hydrogen phosphate	2.3 parts of
		Sodium dihydrogen phosphate	2.3 parts of

Second reagent

Reagent name	Dosage of
		Polyethylene glycol	5 parts of
Sodium azide	7 parts of
		CRP sheep anti-human antibody	2 parts of
Disodium hydrogen phosphate	3 parts of
		Sodium dihydrogen phosphate	3 parts of
Sodium chloride	9 parts of
		Complex protein sodium	6 parts of
Cursor-pull medicine	2 parts of
		Trehalose	3 parts of
Glycerol	2 parts of

Calibration material:

standard dilutions of complement CRP antigen (3 parts citric acid monohydrate, 7 parts sodium chloride, 1 part sodium azide, 2.3 parts sodium hydroxide, 3.7 parts triton, 2 parts disodium hydrogen phosphate, 3 parts sodium dihydrogen phosphate, 5 parts polyvinylpyrrolidone, 3 parts sodium complexing agent,).

Example 2

Complement CRP detection kit

First reagent

Reagent name	Dosage of
		Polyethylene glycol	4 parts of
Sodium azide	5 parts of
		Tween-type oil	3 parts of
Disodium hydrogen phosphate	5 parts of
		Sodium dihydrogen phosphate	6 parts of

Second reagent

Reagent name	Dosage of
		Polyethylene glycol	4 parts of
Sodium azide	5 parts of
		CRP sheep anti-human antibody	7 parts of
Disodium hydrogen phosphate	6 parts of
		Sodium dihydrogen phosphate	8 parts of
Sodium chloride	7 parts of
		Complex protein sodium	5 parts of
Trehalose	1 part of
		Cursor-pull medicine	3 parts of
Glycerol	4 parts of

Calibration material:

standard dilutions of complement CRP antigen (3 parts citric acid monohydrate, 9 parts sodium chloride, 2 parts sodium azide, 2.3 parts sodium hydroxide, 3.7 parts triton, 3 parts disodium hydrogen phosphate, 4 parts sodium dihydrogen phosphate, 6 parts polyvinylpyrrolidone, 3 parts sodium complexing).

Example 3

Complement CRP detection kit

First reagent

Reagent name	Dosage of
		Polyethylene glycol	5 parts of
Sodium azide	5 parts of
		Tween-type oil	4 parts of
Disodium hydrogen phosphate	8 parts of
		Sodium dihydrogen phosphate	7 parts of

Second reagent

Reagent name	Dosage of
		Polyethylene glycol	6 parts of
Sodium azide	11 parts of
		CRP sheep anti-human antibody	8 parts of
Disodium hydrogen phosphate	13 parts of
		Sodium dihydrogen phosphate	11 parts of
Sodium chloride	7 parts of
		Complex protein sodium	9 parts of
Trehalose	5 parts of
		Cursor-pull medicine	2 parts of
Glycerol	3 parts of

Calibration material:

standard dilution for complement CRP antigen (4 parts citric acid monohydrate, 7 parts sodium chloride, 1 part sodium azide, 2.9 parts sodium hydroxide, 3.9 parts triton, 2.4 parts disodium hydrogen phosphate, 3.8 parts sodium dihydrogen phosphate, 5.9 parts polyvinylpyrrolidone, 5 parts sodium collaterals protein)

Example 4

Complement CRP detection kit

First reagent

Reagent name	Dosage of
		Polyethylene glycol	8 parts of
Sodium azide	7 parts of
		Tween-type oil	4 parts of
Disodium hydrogen phosphate	6 parts of
		Sodium dihydrogen phosphate	7 parts of

Second reagent

Calibration material:

standard dilution for complement CRP antigen (3.8 parts citric acid monohydrate, 8 parts sodium chloride, 1.8 parts sodium azide, 2.7 parts sodium hydroxide, 3.7 parts triamcinolone, 4 parts disodium hydrogen phosphate, 5 parts sodium dihydrogen phosphate, 6 parts polyvinylpyrrolidone, 7 parts sodium collaterals protein)

Example 5

Complement CRP detection kit

First reagent

Reagent name	Dosage of
		Polyethylene glycol	15 parts of
Sodium azide	12 parts of
		Tween-type oil	17 parts of
Disodium hydrogen phosphate	13 parts of
		Sodium dihydrogen phosphate	12 parts of

Second reagent

Reagent name	Dosage of
		Polyethylene glycol	15 parts of
Sodium azide	18 parts of
		CRP sheep anti-human antibody	2 parts of
Disodium hydrogen phosphate	3 parts of
		Sodium dihydrogen phosphate	3 parts of
Sodium chloride	9 parts of
		Complex protein sodium	7 parts of
Trehalose	4 parts of
		Cursor-pull medicine	2 parts of
Glycerol	6 parts of

Calibration material:

standard dilution for complement CRP antigen (6 parts citric acid monohydrate, 9 parts sodium chloride, 1 part sodium azide, 3 parts sodium hydroxide, 7 parts triton, 8 parts disodium hydrogen phosphate, 8 parts sodium dihydrogen phosphate, 4 parts polyvinylpyrrolidone, 2 parts sodium collaterals protein)

Comparison of test results in different embodiments

1. One sample with 35mg/L complement CRP concentration was prepared, and the sample was repeatedly tested 7 times with the kit of example 1, and the test 9 results are shown in the table:

it can be seen that the concentration of complement CRP measured in example 1 at 4 months, 6 months and 12 months is close to the true value, and the relative deviation is less than 5%, which indicates that the kit of the invention has good repeatability and accurate measurement.

Example 1 is the optimum formulation ratio.

2. One sample with a CRP concentration of 67.63mg/L was prepared, and the sample was repeatedly tested 7 times with the kit of example 2, and the test 9 results are shown in the table:

/	for 0 month	For 4 months	8 months of	For 12 months	13 months of	For 14 months
							1	67.75	67.94	67.66	67.63	57.71	47.73
2	67.79	67.65	67.98	67.58	57.78	47.57
							3	67.79	67.77	67.82	67.90	57.79	47.97
Theoretical concentration	67.63	67.63	67.63	67.63	67.63	67.63
							Relative deviation 1	0.18％	0.46％	0.04％	0.00％	-14.67％	-29.42％
Relative deviation 2	0.24％	0.03％	0.52％	-0.07％	-14.56％	-29.66％
							Relative deviation 3	0.24％	0.21％	0.28％	0.40％	-14.55％	-29.07％

It can be seen from the table that the concentration of complement CRP measured in example 2 at 4 months, 6 months and 12 months is close to the true value, and the relative deviation is less than 5%, which indicates that the kit of the invention has good repeatability and accurate measurement.

3. One sample with a CRP concentration of 10.00mg/L was prepared, and the sample was repeatedly tested 7 times with the kit of example 3, and the test 9 results are shown in the table:

/	for 0 month	For 4 months	8 months of	For 12 months	13 months of	For 14 months
							1	10.47	10.75	9.93	10.47	8.17	7.14
2	9.98	9.85	10.48	9.82	8.43	7.76
							3	10.78	10.41	10.65	9.59	8.85	7.51
Theoretical concentration	10.00	10.00	10.00	10.00	10.00	10.00
							Relative deviation 1	4.70％	7.50％	-0.70％	4.70％	-18.30％	-28.60％
Relative deviation 2	-0.20％	-1.50％	4.80％	-1.80％	-15.70％	-22.40％
							Relative deviation 3	7.80％	4.10％	6.50％	-4.10％	-11.50％	-24.90％

As can be seen from the table, the concentration of complement CRP measured in example 3 at 1 month, 6 months and 12 months is close to the true value, and the relative deviation is less than 5%, which indicates that the kit of the invention has good repeatability and accurate measurement.

4. One sample with a complement CRP concentration of 9.26mg/L was prepared separately, and the sample was repeatedly tested 7 times with the kit of example 4, with the test 9 results shown in the table:

/	for 0 month	For 4 months	8 months of	For 12 months	13 months of	For 14 months
							1	9.34	8.68	7.90	7.91	6.03	4.61
2	9.22	8.93	7.18	7.59	7.08	5.66
							3	9.34	7.00	7.95	8.40	6.25	7.26
Theoretical concentration	9.26	9.26	9.26	9.26	9.26	9.26
							Relative deviation 1	0.86％	-6.26％	-14.69％	-14.58％	-34.88％	-50.22％
Relative deviation 2	-0.43％	-3.56％	-22.46％	-18.03％	-23.54％	-38.88％
							Relative deviation 3	0.86％	-24.41％	-14.15％	-9.29％	-32.51％	-21.60％

As can be seen from Table 15, the concentration of complement CRP measured at the beginning of 4 months in example 4 all deviate from the true value, and the relative deviation is greater than 15% when left for 4 months, indicating that the kits of the invention have poor reproducibility and inaccurate measurements.

5. One sample with a CRP concentration of 70.2mg/L was prepared, and the sample was repeatedly tested 7 times with the kit of example 5, and the test 9 results are shown in the table:

/	for 0 month	For 4 months	8 months of	For 12 months	13Month of moon	For 14 months
							1	70.41	72.35	63.54	58.29	58.08	55.99
2	70.02	70.89	61.88	57.9	61.93	54.53
							3	71.56	70.91	60.88	56.35	59.14	55.11
Theoretical concentration	70.02	70.02	70.02	70.02	70.02	70.02
							Relative deviation 1	0.56％	3.33％	-9.25％	-16.75％	-17.05％	-20.04％
Relative deviation 2	0.00％	1.24％	-11.63％	-17.31％	-11.55％	-22.12％
							Relative deviation 3	2.20％	1.27％	-13.05％	-19.52％	-15.54％	-21.29％

As can be seen from the table, the concentration of complement CRP measured at 1 month, 6 months and 12 months in example 5 deviates from the true value, and the relative deviation is greater than 15% when left for 8 months, indicating that the kit of the invention has poor reproducibility and inaccurate measurement.

In summary, the bottle opening performance verification of example 2 with the best performance was selected:

it can be seen from the table that the concentration of complement CRP measured in example 1 at 0 to 40 days is close to the true value, and the relative deviation is less than 5%, which indicates that the kit of the invention has good repeatability and accurate measurement.

The above disclosure is only a preferred embodiment of a C-reactive protein assay kit and a detection method with stable performance, but the scope of the invention is not limited thereto, and those skilled in the art will understand that all or part of the procedures for implementing the above embodiments are equivalent and still fall within the scope of the invention.

Claims (5)

1. A C-reactive protein assay kit with stable bottle opening performance is characterized in that,

comprises a first reagent, a second reagent and a standard diluent for complement CRP antigen.

2. A stable C-reactive protein assay kit according to claim 1, wherein,

the first reagent, the second reagent and the standard substance diluent for complement CRP antigen are all preserved at the temperature of 2-8 ℃.

3. A method for detecting a C-reactive protein assay kit with stable bottle opening performance, which is applied to the C-reactive protein assay kit with stable bottle opening performance as claimed in claim 1, and is characterized by comprising the following steps:

adding the first reagent into a sample to be detected, uniformly mixing, incubating at 37 ℃ to obtain a first mixed solution, and reading absorbance A1 of the first mixed solution at a wavelength of 340 nm;

adding the second reagent into the first mixed solution, uniformly mixing, incubating at 37 ℃ to obtain a second mixed solution, and reading absorbance A2 of the second mixed solution at a wavelength of 340 nm;

obtaining the difference value between the absorbance A2 and the absorbance A1 to obtain an absorbance difference;

fitting a standard curve on a full-automatic biochemical analyzer by using the standard substance diluent for the complement CRP antigen, and calculating the content of complement CRP in the sample to be detected based on the absorbance difference.

4. The C-reactive protein assay kit and the assay method for stable performance in opening bottles of claim 3, wherein,

the volume ratio of the sample to be tested to the first reagent is 17:300, the volume ratio of the second reagent to the first reagent is 1:4.

5. a method for preparing a C-reactive protein assay kit with stable opening performance, for preparing a C-reactive protein assay kit with stable opening performance according to claim 1, comprising the steps of:

obtaining polyethylene glycol, sodium azide, tween, purified water, disodium hydrogen phosphate and sodium dihydrogen phosphate, uniformly mixing at 25+/-3 ℃ and filtering to obtain a first reagent;

obtaining water-soluble sodium channel, triton, trehalose, glycerol, sodium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium azide, citric acid, CRP sheep anti-human antibody and purified water, uniformly mixing at 25+/-3 ℃ and filtering to obtain a second reagent;

obtaining citric acid monohydrate, sodium chloride, sodium azide, sodium hydroxide, triton, disodium hydrogen phosphate, sodium dihydrogen phosphate, polyvinylpyrrolidone, sodium complexing agent and purified water, and diluting the citric acid monohydrate, sodium chloride, sodium azide, sodium hydroxide, triton, sodium complexing agent and purified water into complement CRP calibrator with different concentrations by using a calibrator to obtain a standard diluent for complement CRP antigen;

and packaging the first reagent, the second reagent and the complement CRP antigen with a standard substance diluent to obtain a kit.