A novel immunoturbidimetric kit for detecting procalcitonin
By using polystyrene latex microspheres coupled with monoclonal antibody combination antibodies in the procalcitonin immunoturbidimetry kit, combined with appropriate ratios and processes, the low response and false positive problems caused by multiple antibodies are solved. , significantly improves the accuracy and stability of the kit.
Patent Information
- Application Number
- CN202210418939.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Priority Date
- 2021-12-22
- Filing Date
- 2022-04-20
- Publication Date
- 2024-12-31
- Estimated Expiration
- 2042-04-20
AI Technical Summary
The existing procalcitonin immunoturbidity kits have low reaction levels and false positives due to the use of multiple antibodies, especially when the absorbance is low near the cut-off value.
Polystyrene latex microspheres coupled with monoclonal antibody combination antibody, combined with suitable buffers and alkali metal salts, and prepared a novel immunologic turbidity for detecting procalcitonin through specific ratios and process preparation methods. Kits to reduce batch difference and improve reaction.
It significantly improves the turbidity reaction of the kit, reduces false positives, and has a much higher accuracy than that of the kit using polyclonal antibodies. Through the use of CE510 blocking agent and tea saponin, the stability and credibility have been improved. .
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Figure CN114778824B_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of detection reagents, and in particular relates to a novel immunoturbidimetric kit for detecting procalcitonin. Background Art
[0002] Immunoturbidimetric kits are generally used in medical testing. The principle of latex-enhanced immunoturbidimetry is that the target in the sample binds to the antibodies on the surface of the latex particles. The higher the concentration of the target in the sample, the greater the degree of binding of the antibodies on the latex surface, and the higher the degree of latex aggregation that can be induced. The turbidity of the reaction system is positively correlated with the concentration of the target in the sample. By measuring the absorbance at a specific wavelength, a calibration curve is established to calculate the concentration of the target.
[0003] Currently, the antibodies coupled to the surface of latex microspheres are polyclonal antibodies. The higher the procalcitonin concentration in the sample, the greater the degree of binding of the antibodies on the latex surface, and the higher the degree of latex aggregation that can be induced. The turbidity of the reaction system is positively correlated with the procalcitonin concentration in the sample. By measuring the absorbance at 600nm, a calibration curve is established to calculate the procalcitonin concentration in serum or plasma samples.
[0004] The existing antibodies are polyclonal antibodies with large batch differences; there are few target antibodies for antigen epitopes, and there are mixed antibodies, which leads to low absorbance near the cut-off value and easy false positive phenomenon; the antibodies used in the calcitonin pro-immunoturbidimetric kit on the market are all polyclonal antibodies, and the reactivity of monoclonal antibodies in immunoturbidimetric analysis is low. It is necessary to find several monoclonal antibodies to combine with sensitized latex microspheres to solve the problem of low reactivity of monoclonal antibodies in immunoturbidimetric analysis. Summary of the invention
[0005] The object of the present invention is to provide a novel immunoturbidimetric kit for detecting procalcitonin to solve the problems in the background technology.
[0006] The purpose of the present invention can be achieved by the following technical scheme: A novel immunoturbidimetric kit for detecting procalcitonin, comprising a procalcitonin standard, R1 reagent and R2 reagent, wherein the R1 reagent and the R2 reagent comprise the following components according to the ratio:
[0007] R1 reagent: 0.4-0.6wt% buffer, 0.08-0.12wt% bovine serum albumin, 0.8-1.2wt% alkali metal salt, 0.08-0.12wt% D-trehalose, 0.5-3.0wt% coagulant, 0.1-1.0wt% tea saponin, 0.08-0.12wt% sodium azide, and the balance is deionized water;
[0008] R2 reagent: 48-52mmol / L buffer, 0.08-0.12vol% CE510 blocking agent, 0.8-1.2wt% alkali metal salt, 0.08-0.12wt% D-trehalose, 0.08-0.12wt% sodium azide, 0.08-0.12wt% mouse anti-human procalcitonin monoclonal antibody-coupled polystyrene latex microspheres, the balance is deionized water;
[0009] Furthermore, procalcitonin standards were purchased from TRC (Toronto Research Chemicals);
[0010] Further, the buffer in the R1 reagent and the R2 reagent is one or more of 2-(N-morpholino)ethanesulfonic acid monohydrate-NaOH, 4-hydroxyethylpiperazineethanesulfonic acid-HCl, tris(hydroxymethyl)aminomethane, N-2-hydroxyethylpiperazine-N-3-propanesulfonic acid, 3-(N-morphinogen)propanesulfonic acid, triethanolamine, diethanolamine, 2-methyl-2-amino-1-propanol, N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, (cyclohexylamine)-1-propanesulfonic acid, (cyclohexylamine)-1-ethanesulfonic acid, 2-(N-morpholino)ethanesulfonic acid, piperazine-N,N-bis(2-ethanesulfonic acid), piperazine-N,N-bis(2-hydroxyethanesulfonic acid), tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid, and tris(hydroxymethyl)methyl-2-aminosulfonic acid, mixed in any ratio;
[0011] Further, the pH value of the R1 reagent is 5.1-7.4, and the coagulant is polyethylene glycol;
[0012] Further, the pH value of the R2 reagent is 7.4-7.8, the concentration of the mouse anti-human procalcitonin monoclonal combination antibody in the polystyrene latex microspheres coupled with the mouse anti-human procalcitonin monoclonal combination antibody is 0.02-0.09 mg / mL, and the final concentration of the latex microspheres is 0.09-0.13 wt %; the CE510 blocking agent is purchased from JSR;
[0013] Further, a mouse anti-human procalcitonin monoclonal combination antibody was prepared by mixing monoclonal antibody PCT-124E7, monoclonal antibody PCT-11B8 and monoclonal antibody PCT-37G5 at a mass ratio of 1:1:1;
[0014] Further, the alkali metal salt is one or more of NaCl, KCl, Na2HPO3, NaH2PO3, K2HPO3, KH2PO3, Na2CO3, NaHCO3 and K2CO3 mixed in any ratio;
[0015] Furthermore, the mouse anti-human procalcitonin monoclonal antibody-coupled polystyrene latex microspheres are prepared by the following steps:
[0016] The polystyrene latex microspheres containing carboxyl groups on the surface were mixed with a MES buffer having a pH value of 6.0 and a concentration of 0.01 mol / L, and then EDAC / NHS was added for activation after being blown evenly with a pipette. The mixture was reacted in a constant temperature shaker at 25°C for 15-20 minutes, and the supernatant was removed after centrifugation at a centrifugal force of 15000g for 30 minutes. The precipitate was resuspended in a 4-hydroxyethylpiperazineethanesulfonic acid buffer having a pH value of 7.6 and a concentration of 25 mmol / L to obtain a suspension, and the activated polystyrene latex microspheres containing carboxyl groups on the surface were obtained by ultrasonic treatment. polystyrene latex microspheres; adding mouse anti-human procalcitonin monoclonal combination antibody to the suspension after ultrasonic treatment, reacting in a constant temperature shaker at 25°C for 1-2 hours, then adding BSA glycine buffer with a concentration of 10g / L, reacting in a constant temperature shaker at 25°C for 40 minutes, centrifuging for 30 minutes under the condition of a centrifugal force of 13000g, resuspending the precipitate twice with R2 buffer, centrifuging and storing the precipitate in R2 buffer treated with ultrasonic treatment for 5 minutes, and obtaining mouse anti-human procalcitonin monoclonal combination antibody-coupled polystyrene latex microspheres;
[0017] The dosage ratio of polystyrene latex microspheres containing carboxyl groups on the surface, MES buffer, EDAC / NHS, 4-hydroxyethylpiperazineethanesulfonic acid buffer, mouse anti-human procalcitonin monoclonal combination antibody, BSA glycine buffer, R2 buffer and R2 buffer treated with ultrasound for 5 min is 0.1mL: 0.9mL: 0.2mL: 1mL: 0.1mg: 2mL: 1mL: 1mL; the mass ratio of EDAC and NHS in EDAC / NHS is 1:2;
[0018] Furthermore, the average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is preferably 100-400 nm;
[0019] Further, the surface carboxyl density of polystyrene latex microspheres containing carboxyl groups on the surface is preferably 0.03-0.30 mmol / g and is purchased from JSR Life Sciences;
[0020] Furthermore, on the basis of the R2 reagent, the polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combined antibody are not added, and the remaining components and the dosage ratio of each component remain unchanged to obtain the R2 buffer solution.
[0021] Beneficial effects of the present invention:
[0022] R1 and R2 reagents contain alkali metal salts, which make the reagent solution have appropriate ionic strength, which is conducive to the stable storage of latex microspheres coupled with anti-target detection antibodies and target detection objects; CE510 blocking agent in R2 reagent can occupy the blank sites on latex microspheres to prevent latex microspheres from agglutination; adding a certain concentration of tea saponin to R1 reagent, because tea saponin is a stronger surfactant and has the function of dissolving lipids, can eliminate the different degrees of interference of different sample detection reagents, and will not interfere with the binding of monoclonal antibodies to the target, thereby ensuring the credibility of the test results. The kit uses monoclonal antibodies, which can reduce the batch difference of the kit.
[0023] Through the latex-enhanced immunoturbidimetric detection test, it was found that the turbidimetric reactivity of the kit was high and there was no false positive phenomenon. The labeled combination monoclonal antibody used in the latex-enhanced immunoturbidimetric detection kit was highly consistent with the gold standard Roche electrochemiluminescence detection value, and its accuracy was much higher than that of the existing labeled polyclonal antibody kit. BRIEF DESCRIPTION OF THE DRAWINGS
[0024] The present invention will be further described below in conjunction with the accompanying drawings.
[0025] Figure 1 This is a correlation diagram of the clinical comparison between the labeled polyclonal antibody of the present invention and Roche's electrochemiluminescence;
[0026] Figure 2 It is a correlation diagram of the clinical comparison between the labeled combination monoclonal antibody of the present invention and Roche's electrochemiluminescence. DETAILED DESCRIPTION
[0027] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0028] Example 1
[0029] Preparation of mouse anti-human procalcitonin monoclonal antibody-coupled polystyrene latex microspheres comprises the following steps:
[0030] Mouse anti-human procalcitonin monoclonal combination antibody was prepared by mixing monoclonal antibody PCT-124E7, monoclonal antibody PCT-11B8 and monoclonal antibody PCT-37G5 at a mass ratio of 1:1:1;
[0031] The average particle size of polystyrene latex microspheres containing carboxyl groups on the surface is 100 nm and the density is 0.03 mmol / g;
[0032] 0.1 mL of polystyrene latex microspheres containing carboxyl groups on the surface were mixed with 0.9 mL of MES buffer with a pH value of 6.0 and a concentration of 0.01 mol / L, and then 0.2 mL of EDAC / NHS was added for activation after being blown evenly with a pipette, and the mixture was reacted in a constant temperature shaker at 25°C for 15 min, and the supernatant was removed after centrifugation at a centrifugal force of 15000 g for 30 min. The precipitate was resuspended with 1 mL of 4-hydroxyethylpiperazineethanesulfonic acid buffer with a pH value of 7.6 and a concentration of 25 mmol / L to obtain a suspension, and ultrasonic treatment was performed to obtain activated polystyrene latex microspheres containing carboxyl groups on the surface; 0.1 mg of mouse anti-human procalcitonin monoclonal combination antibody was added to the suspension after ultrasonic treatment, and the mixture was reacted in a constant temperature shaker at 25°C for 1 h, and then 1 mL of BSA glycine buffer with a concentration of 10 g / L was added, and the mixture was reacted in a constant temperature shaker at 25°C for 40 min, and the mixture was centrifuged at a centrifugal force of 13000 g for 30 min, and the precipitate was resuspended with 2 mL The mixture was resuspended twice in R2 buffer, and the precipitate was collected by centrifugation and stored in 1 mL of R2 buffer treated with ultrasound for 5 min to obtain polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combination antibodies; the mass ratio of EDAC to NHS in EDAC / NHS was 1:2.
[0033] Example 2
[0034] Preparation of mouse anti-human procalcitonin monoclonal antibody-coupled polystyrene latex microspheres comprises the following steps:
[0035] Mouse anti-human procalcitonin monoclonal combination antibody was prepared by mixing monoclonal antibody PCT-124E7, monoclonal antibody PCT-11B8 and monoclonal antibody PCT-37G5 at a mass ratio of 1:1:1;
[0036] The average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is preferably 400 nm and the density is 0.30 mmol / g;
[0037] 0.1 mL of polystyrene latex microspheres containing carboxyl groups on the surface were mixed with 0.9 mL of MES buffer with a pH value of 6.0 and a concentration of 0.01 mol / L, and then 0.2 mL of EDAC / NHS was added for activation, and the mixture was reacted in a constant temperature shaker at 25°C for 20 min. After centrifugation at a centrifugal force of 15000 g for 30 min, the supernatant was removed, and the precipitate was resuspended with 1 mL of 4-hydroxyethylpiperazineethanesulfonic acid buffer with a pH value of 7.6 and a concentration of 25 mmol / L to obtain a suspension, and ultrasonic treatment was performed to obtain activated polystyrene latex microspheres containing carboxyl groups on the surface; 0.1 mg of mouse anti-human procalcitonin monoclonal combination antibody was added to the suspension after ultrasonic treatment, and the mixture was reacted in a constant temperature shaker at 25°C for 2 h, and then 1 mL of BSA glycine buffer with a concentration of 10 g / L was added, and the mixture was reacted in a constant temperature shaker at 25°C for 40 min. After centrifugation at a centrifugal force of 13000 g for 30 min, the precipitate was resuspended with 2 mL The mixture was resuspended twice in R2 buffer, and the precipitate was collected by centrifugation and stored in 1 mL of R2 buffer treated with ultrasound for 5 min to obtain polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combination antibodies; the mass ratio of EDAC to NHS in EDAC / NHS was 1:2.
[0038] Example 3
[0039] Preparation of mouse anti-human procalcitonin monoclonal antibody-coupled polystyrene latex microspheres comprises the following steps:
[0040] Mouse anti-human procalcitonin monoclonal combination antibody was prepared by mixing monoclonal antibody PCT-124E7, monoclonal antibody PCT-11B8 and monoclonal antibody PCT-37G5 at a mass ratio of 1:1:1;
[0041] The average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is preferably 270 nm and the density is 0.134 mmol / g;
[0042] 0.1 mL of polystyrene latex microspheres containing carboxyl groups on the surface were mixed with 0.9 mL of MES buffer with a pH value of 6.0 and a concentration of 0.01 mol / L, and then 0.2 mL of EDAC / NHS was added for activation, and the mixture was reacted in a constant temperature shaker at 25°C for 20 min. After centrifugation at a centrifugal force of 15000 g for 30 min, the supernatant was removed, and the precipitate was resuspended with 1 mL of 4-hydroxyethylpiperazineethanesulfonic acid buffer with a pH value of 7.6 and a concentration of 25 mmol / L to obtain a suspension, and ultrasonic treatment was performed to obtain activated polystyrene latex microspheres containing carboxyl groups on the surface; 0.1 mg of mouse anti-human procalcitonin monoclonal combination antibody was added to the suspension after ultrasonic treatment, and the mixture was reacted in a constant temperature shaker at 25°C for 2 h, and then 1 mL of BSA glycine buffer with a concentration of 10 g / L was added, and the mixture was reacted in a constant temperature shaker at 25°C for 40 min. After centrifugation at a centrifugal force of 13000 g for 30 min, the precipitate was resuspended with 2 mL The mixture was resuspended twice in R2 buffer, and the precipitate was collected by centrifugation and stored in 1 mL of R2 buffer treated with ultrasound for 5 min to obtain polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combination antibodies; the mass ratio of EDAC to NHS in EDAC / NHS was 1:2.
[0043] Example 4
[0044] The composition of the novel immunoturbidimetric kit for detecting procalcitonin is shown in Table 1 below:
[0045] Table 1
[0046]
[0047] The concentration of mouse anti-human procalcitonin monoclonal combined antibody in the polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combined antibody was 0.02 mg / mL, and the final concentration of the latex microspheres was 0.09 wt %.
[0048] Example 5
[0049] The composition of the novel immunoturbidimetric kit for detecting procalcitonin is shown in Table 2 below:
[0050] Table 2
[0051]
[0052] The concentration of mouse anti-human procalcitonin monoclonal combined antibody in the polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combined antibody was 0.05 mg / mL, and the final concentration of the latex microspheres was 0.11 wt %.
[0053] Example 6
[0054] The composition of the novel immunoturbidimetric kit for detecting procalcitonin is shown in Table 3 below:
[0055] Table 3
[0056]
[0057] The concentration of mouse anti-human procalcitonin monoclonal combined antibody in the polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combined antibody was 0.09 mg / mL, and the final concentration of the latex microspheres was 0.13 wt %.
[0058] Example 7
[0059] The composition of the novel immunoturbidimetric kit for detecting procalcitonin is shown in Table 4 below:
[0060] Table 4
[0061]
[0062] The concentration of mouse anti-human procalcitonin monoclonal combined antibody in the polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combined antibody was 0.03 mg / mL, and the final concentration of the latex microspheres was 0.1 wt %.
[0063] The latex enhanced immunoturbidimetric test was performed using the novel immunoturbidimetric kit for detecting procalcitonin in Example 7. First, parameters (Hitachi 7180 automatic biochemical analyzer) were set with reference to Table 5.
[0064] Table 5
[0065]
[0066] Mix 15 μl of procalcitonin standard with reagent R1, incubate at 37°C for 5 minutes, then mix with 60 μl of reagent R2, incubate at 37°C for about 20 seconds, read the absorbance value A1, incubate for 5 minutes, read the absorbance value A2, calculate ΔA=A2-A1, draw a standard curve, and calculate the procalcitonin content in the sample according to the standard curve. The process is shown in Table 6.
[0067] Table 6
[0068]
[0069] The test results are shown in Table 7 and Figure 1 and Figure 2 .
[0070] Table 7
[0071]
[0072]
[0073]
[0074] From Table 7, Figure 1 and Figure 2 It can be seen that the labeled combination monoclonal antibody used in the latex-enhanced immunoturbidimetric detection kit is highly consistent with the gold standard Roche electrochemiluminescence detection value, and its accuracy is much higher than that of the existing labeled polyclonal antibody kit, and no false positive phenomenon occurs near the cut-off value.
[0075] It should be noted that, in this article, terms such as "comprises", "includes" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or apparatus.
[0076] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A novel immunoturbidimetric kit for detecting procalcitonin, characterized in that: It includes procalcitonin standard, R1 reagent and R2 reagent. R1 reagent and R2 reagent include the following components according to the proportion: R1 reagent: 0.4-0.6wt% buffer, 0.08-0.12wt% bovine serum albumin, 0.8-1.2wt% alkali metal salt, 0.08-0.12wt% D-trehalose, 0.5-3.0wt% coagulant, 0.1-1.0wt% tea saponin, 0.08-0.12wt% sodium azide, and the balance is deionized water; R2 reagent: 48-52mmol / L buffer, 0.08-0.12vol% CE510 blocking agent, 0.8-1.2wt% alkali metal salt, 0.08-0.12wt% D-trehalose, 0.08-0.12wt% sodium azide, 0.08-0.12wt% mouse anti-human procalcitonin monoclonal antibody-coupled polystyrene latex microspheres, the balance is deionized water; The alkali metal salt is one or more of NaCl, KCl, Na2HPO3, NaH2PO3, K2HPO3, KH2PO3, Na2CO3, NaHCO3 and K2CO3 mixed in any ratio; The mouse anti-human procalcitonin monoclonal combination antibody is prepared by mixing monoclonal antibody PCT-124E7, monoclonal antibody PCT-11B8 and monoclonal antibody PCT-37G5 in a mass ratio of 1:1:
1.
2. A novel immunoturbidimetric kit for detecting procalcitonin according to claim 1, characterized in that: The pH value of the R1 reagent is 5.1-7.4, and the coagulant is polyethylene glycol.
3. A novel immunoturbidimetric kit for detecting procalcitonin according to claim 1, characterized in that: The pH value of R2 reagent is 7.4-7.
8.
4. A novel immunoturbidimetric kit for detecting procalcitonin according to claim 1, characterized in that: The buffer in reagent R1 and reagent R2 is one or more of 2-(N-morpholino)ethanesulfonic acid monohydrate-NaOH, 4-hydroxyethylpiperazineethanesulfonic acid-HCl, tris(hydroxymethyl)aminomethane, N-2-hydroxyethylpiperazine-N-3-propanesulfonic acid, 3-(N-morpholino)propanesulfonic acid, triethanolamine, diethanolamine, 2-methyl-2-amino-1-propanol, N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, (cyclohexylamine)-1-propanesulfonic acid, (cyclohexylamine)-1-ethanesulfonic acid, 2-(N-morpholino)ethanesulfonic acid, piperazine-N,N-bis(2-ethanesulfonic acid), piperazine-N,N-bis(2-hydroxyethanesulfonic acid), tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid, and tris(hydroxymethyl)methyl-2-aminosulfonic acid mixed in any ratio.
5. A novel immunoturbidimetric kit for detecting procalcitonin according to claim 1, characterized in that: Mouse anti-human procalcitonin monoclonal antibody-coupled polystyrene latex microspheres were prepared by the following steps: The polystyrene latex microspheres containing carboxyl groups on the surface are mixed with MES buffer, blown evenly, and then EDAC / NHS is added for activation, reacted at 25°C for 15-20 minutes, centrifuged for 30 minutes, and then the precipitate is resuspended with 4-hydroxyethylpiperazineethanesulfonic acid buffer with a pH value of 7.6 and a concentration of 25mmol / L to obtain a suspension, and ultrasonic treatment is performed to obtain activated polystyrene latex microspheres containing carboxyl groups on the surface; mouse anti-human procalcitonin monoclonal combination antibody is added to the suspension after ultrasonic treatment, and the reaction is carried out in a constant temperature shaker at 25°C for 1-2 hours, and then BSA glycine buffer with a concentration of 10g / L is added, and the reaction is carried out at 25°C for 40 minutes, and centrifuged at a centrifugal force of 13000g for 30 minutes, and the precipitate is resuspended twice with R2 buffer, and the precipitate is centrifuged and stored in R2 buffer treated with ultrasonic treatment for 5 minutes to obtain polystyrene latex microspheres coupled with mouse anti-human procalcitonin monoclonal combination antibody.
6. A novel immunoturbidimetric kit for detecting procalcitonin according to claim 5, characterized in that: The dosage ratio of polystyrene latex microspheres containing carboxyl groups on the surface, MES buffer, EDAC / NHS, 4-hydroxyethylpiperazineethanesulfonic acid buffer, mouse anti-human procalcitonin monoclonal combination antibody, BSA glycine buffer, R2 buffer and R2 buffer treated with ultrasound for 5 min is 0.1mL:0.9mL:0.2mL:1mL:0.1mg:2mL:1mL:1mL; the mass ratio of EDAC and NHS in EDAC / NHS is 1:
2.
7. A novel immunoturbidimetric kit for detecting procalcitonin according to claim 5, characterized in that: The average particle size of the polystyrene latex microspheres containing carboxyl groups on the surface is 100-400nm.
8. A novel immunoturbidimetric kit for detecting procalcitonin according to claim 7, characterized in that: The surface carboxyl density of the polystyrene latex microspheres containing carboxyl groups on the surface is 0.03-0.30 mmole / g.