A homogeneous chemiluminescence assay kit for procalcitonin

Abstract

The present invention belongs to the field of in vitro detection technology, and in particular to a homogeneous chemiluminescence assay kit for procalcitonin. The procalcitonin test kit of the present invention includes R1 reagent and R2 reagent independently of each other, wherein the R1 reagent includes luminescent particles coated with procalcitonin monoclonal antibody, surfactant A and stabilizer, and the R2 reagent includes biotin-labeled procalcitonin monoclonal antibody and surfactant B, wherein the surfactant A includes a mixture of Tween 80 and PVP K30 in a mass ratio of 1-2:1, and the surfactant B includes a mixture of Triton x-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:2-4. After adding surfactant A and surfactant B in a specific mass ratio, the present invention can exert a good component enhancement effect; by reasonably controlling the content of each substance in R1 reagent and R2, the accuracy, precision, stability and specificity of the procalcitonin assay kit are improved, and the linear relationship is good, which meets clinical needs.

Description

Technical Field

[0001] The invention belongs to the technical field of in vitro detection, and in particular relates to a homogeneous chemiluminescence assay kit for procalcitonin. Background Art

[0002] Procalcitonin (PCT) is a calcitonin propeptide substance without hormonal activity, which comes from a single-copy gene located on chromosome 11. The procalcitonin precursor generates 116 amino acids of PCT with a molecular weight of about 13kD under the action of endogenous peptidases. The concentration of PCT in healthy individuals is very low, and it is a very stable protein both in vivo and in vitro. When a systemic bacterial infection occurs, PCT can be ectopically produced and released into the blood circulation, and can be detected early after infection. The detection of PCT can be used as an indication for the initiation of antibiotic treatment, and dynamic monitoring of PCT levels can assist in evaluating the therapeutic effect of antibiotics. In addition, when there is severe non-infectious inflammatory stimulation, such as large-area burns, severe trauma, acute multiple organ failure and cardiac surgery, PCT can also be used as one of the evaluation indicators for early diagnosis.

[0003] At present, the main methods used to detect PCT are radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), colloidal gold immunochromatography (GICA) and chemiluminescent immunoassay (CLIA). RIA has high detection sensitivity, but due to the radioactive hazards of the marker, poor stability of the marker, and difficulty in handling waste, it has gradually withdrawn from the field of clinical testing; ELISA uses horseradish peroxidase or alkaline phosphatase to label antibodies and catalyze the substrate to produce color changes. It has the characteristics of simple operation and long reagent stability period, but the detection sensitivity of ELISA is low. It is currently mainly used in projects with low detection sensitivity requirements such as infectious disease screening; GICA has the advantages of simple operation and fast detection speed, but it also has the disadvantages of low sensitivity, unstable reagents, poor repeatability, and difficulty in quantification; CLIA is an immunoassay technology developed on the basis of enzyme-linked immunosorbent assay, which has the advantages of high sensitivity, wide detection linear range, simple operation, and high degree of automation.

[0004] Chinese invention patent CN103901203A discloses a chemiluminescent quantitative detection kit for procalcitonin and its preparation method and detection method. The kit includes the following reagents: a series of procalcitonin standards; a magnetic separation reagent: a luminescent microparticle suspension coupled with streptavidin; a first reagent containing an anti-procalcitonin monoclonal antibody solution labeled with N-hydroxysuccinimide biotin ester; and a second reagent containing an anti-procalcitonin monoclonal antibody solution labeled with alkaline phosphatase. The invention uses a luminescent microparticle suspension coupled with streptavidin, an anti-procalcitonin monoclonal antibody solution labeled with N-hydroxysuccinimide biotin ester, and an anti-procalcitonin monoclonal antibody solution labeled with alkaline phosphatase to prepare a kit, so that the sensitivity reaches 0.008 ng / mL. Although the sensitivity is improved, the specificity of the kit is poor and the detection range is general (0100 ng / mL).

[0005] Another Chinese invention patent CN102359958A discloses a PCT chemiluminescence detection kit, which uses luminescent microparticles coupled with antibodies as the fixed separation phase, a biotin-streptavidin multi-stage amplification system, and horseradish peroxidase as the catalytic enzyme to catalyze the luminescent substrate to emit light, but its detection sensitivity is only 0.05ng / mL.

[0006] In view of this, there is an urgent need in the art for a procalcitonin assay kit with high precision, strong specificity and wide detection range. Summary of the invention

[0007] The invention aims at the problems existing in the prior art and provides a homogeneous chemiluminescence assay kit for procalcitonin.

[0008] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0009] A procalcitonin assay kit comprises an R1 reagent and an R2 reagent which are independent of each other, wherein the R1 reagent comprises luminescent microparticles coated with a procalcitonin monoclonal antibody, a surfactant A and a stabilizer, and the R2 reagent comprises a biotin-labeled procalcitonin monoclonal antibody and a surfactant B, wherein the surfactant A comprises a mixture of Tween 80 and PVPK30 in a mass ratio of 1-2:1, and the surfactant B comprises a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:2-4.

[0010] Preferably, the surfactant A comprises a mixture of Tween 80 and PVP K30 in a mass ratio of 1-1.5:1, and the surfactant B comprises a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:2-3.

[0011] Preferably, the stabilizer comprises a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 2-10:1-4:1.

[0012] More preferably, the stabilizer comprises a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 2-5:1-3:1.

[0013] Preferably, the procalcitonin assay kit further comprises streptavidin-coated photosensitive microspheres, the R1 reagent comprises a buffer solution, and the R2 reagent comprises a permeabilizing agent and a preservative.

[0014] Preferably, the buffer is selected from one or more of a phosphate buffer, a Tris buffer, a MOPS buffer, a PBS buffer, a MES buffer and a glycine buffer.

[0015] Preferably, the pH of the buffer solution is 6.5-7.5.

[0016] Preferably, the osmotic agent is selected from one or more of sodium chloride, β-cyclodextrin and carboxymethyl cellulose.

[0017] Preferably, the preservative is selected from one or more of phenol, sodium benzoate and sodium azide.

[0018] Preferably, the procalcitonin assay kit comprises R1 reagent and R2 reagent which are independent of each other, wherein the R1 reagent comprises 30-40 mg / L of luminescent microparticles coated with procalcitonin monoclonal antibody, 2-3 g / L of surfactant A, 0.5-1 g / L of stabilizer and 4-6 g / L of buffer, and the R2 reagent comprises 6-10 mg / L of biotin-labeled procalcitonin monoclonal antibody, 2-4 mL / L of surfactant B, 4-6 g / L of osmotic agent and 1-1.5 mL / L of preservative.

[0019] Preferably, the method of using the procalcitonin assay kit is to perform assay using an automatic chemiluminescence immunoassay using an endpoint method, with the dominant detection wavelength being 680 nm, wherein the volume ratio of the R1 reagent to the R2 reagent is 1-2:1.

[0020] The present invention also provides the use of the procalcitonin determination kit in determining procalcitonin in serum for non-diagnostic purposes.

[0021] The procalcitonin assay kit of the present invention adopts the principle of double antibody sandwich immunoassay, which is a homogeneous immune reaction.

[0022] The first incubation: a certain amount of the sample to be tested is added, and a PCT monoclonal antibody-coated luminescent microsphere and a biotin-labeled PCT monoclonal antibody are incubated together to react and form an antibody-antigen-antibody sandwich complex.

[0023] Second incubation: After adding streptavidin-coated photosensitive microspheres, the complex specifically binds to them through the interaction between biotin and streptavidin.

[0024] In a homogeneous environment, under 680nm laser excitation conditions, singlet oxygen was transferred between the particles, eventually generating a 615nm light signal. The light signal value was measured by a single-photon counter and then the final detection result was obtained through the calibration curve of the detector. The calibration curve was calculated by a 6-point calibration fit, and the total detection time was 12 minutes.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] (1) During the implementation of the present invention, it was unexpectedly found that when surfactant A composed of Tween 80 and PVP K30 in a specific mass ratio and surfactant B composed of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a specific mass ratio are present in R1 reagent and R2 reagent at a certain content, a relatively good enhancement effect can be exerted.

[0027] (2) The present invention adds a stabilizer to the R1 reagent, namely bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 2-10:1-4:1, thereby improving the stability of the procalcitonin assay kit while reducing the amount of preservative added.

[0028] (3) The present invention improves the accuracy, precision, stability and specificity of the procalcitonin assay kit by reasonably controlling the content of each substance in the R1 reagent and the R2 reagent, and has a good linear relationship, which meets clinical needs. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is the linear range of the procalcitonin assay kit of Example 1 of the present invention. DETAILED DESCRIPTION

[0030] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further explained in conjunction with specific examples below, but the following examples are only preferred embodiments of the present invention, not all. Based on the examples in the implementation mode, other embodiments obtained by those skilled in the art without creative work all belong to the protection scope of the present invention. The experimental methods in the following examples, unless otherwise specified, are conventional methods, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0031] Among them, the luminescent particles (Dynabeads TMM-280Tosylactivated, catalog number 14204, purchased from Thermo Fisher Scientific Inc.; procalcitonin monoclonal antibody, catalog number C7113-100UL, purchased from Sigma; biotin (EZ-Link TM NHS, catalog number 20217, purchased from Thermo Fisher Scientific Inc.; pure procalcitonin, catalog number P7010, purchased from Guangzhou Greenline Biotechnology Co., Ltd.; streptavidin-coated photosensitive microspheres (Power-Bind TM ), catalog number 29000701010150, purchased from Thermo Fisher Scientific Inc.; bovine serum albumin, CAS number: 9048-46-8, purchased from Aladdin.

[0032] The preparation method of luminescent microparticles coated with procalcitonin monoclonal antibody is as follows: take an appropriate amount of luminescent microparticles, add them to 10 mL of buffer solution, so that the final concentration of the luminescent microparticles is 1.0%; then add an appropriate amount of procalcitonin monoclonal antibody, mix and stir at room temperature for about 3 hours, add 1 ml of 10 g / L bovine serum albumin solution, centrifuge at 12000 rpm for 40 minutes, remove the supernatant, and the resulting precipitate is the luminescent microparticles coated with procalcitonin monoclonal antibody.

[0033] Preparation method of biotin-labeled procalcitonin monoclonal antibody: add procalcitonin monoclonal antibody to a centrifuge tube with a filter membrane, centrifuge at 8000r / min for 6min, wash repeatedly with buffer 3-5 times, add 5μL 22mg / mL biotin (prepared with DMSO) to 50μL antibody solution, incubate at room temperature for 4h, dialyze at 4℃ for 24h, change the liquid once every 6h, collect the liquid in the dialysis bag, dilute and use it for later use.

[0034] Example 1

[0035] A procalcitonin assay kit comprises a mycoavidin-coated photosensitive microsphere, and R1 reagent and R2 reagent which are independent of each other.

[0036] The R1 reagent consists of the following components:

[0037] 35 mg / L of luminescent microparticles coated with procalcitonin monoclonal antibody, 2.5 g / L of surfactant A (a mixture of Tween 80 and PVP K30 in a mass ratio of 1:1), 0.8 g / L of stabilizer (a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 3:2:1), and 5 g / L of glycine buffer (pH = 6.5).

[0038] R2 reagent consists of the following components:

[0039] Biotin-labeled procalcitonin monoclonal antibody 8 mg / L, surfactant B 3 ml / L (a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:3), sodium chloride 5 g / L, sodium azide 1 mL / L.

[0040] Example 2

[0041] A procalcitonin assay kit comprises a photosensitive microsphere coated with streptavidin, and R1 reagent and R2 reagent which are independent of each other.

[0042] The R1 reagent consists of the following components:

[0043] 30 mg / L of luminescent microparticles coated with procalcitonin monoclonal antibody, 2 g / L of surfactant A (a mixture of Tween 80 and PVP K30 in a mass ratio of 2:1), 0.5 g / L of stabilizer (a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 2:1:1), and 4 g / L of Tris buffer (pH = 7.5).

[0044] R2 reagent consists of the following components:

[0045] Biotin-labeled procalcitonin monoclonal antibody 6 mg / L, surfactant B 2 ml / L (a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:2), carboxymethyl cellulose 6 g / L, sodium benzoate 1.2 mL / L.

[0046] Example 3

[0047] A procalcitonin assay kit comprises a mycoavidin-coated photosensitive microsphere, and R1 reagent and R2 reagent which are independent of each other.

[0048] The R1 reagent consists of the following components:

[0049] 40 mg / L of luminescent microparticles coated with procalcitonin monoclonal antibody, 3 g / L of surfactant A (a mixture of Tween 80 and PVP K30 in a mass ratio of 1:1), 1 g / L of stabilizer (a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 10:4:1), and 6 g / L of phosphate buffer (pH = 7).

[0050] R2 reagent consists of the following components:

[0051] Biotin-labeled procalcitonin monoclonal antibody 10 mg / L, surfactant B 4 ml / L (a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:4), β-cyclodextrin 4 g / L, sodium azide 1.5 mL / L.

[0052] Comparative Example 1

[0053] The only difference from Example 1 is that the stabilizer is only a mixture of gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 4:2, and the other components and concentrations are the same as those in Example 1.

[0054] Comparative Example 2

[0055] The only difference from Example 1 is that the stabilizer is only bovine serum albumin, and the other components and concentrations are the same as those in Example 1.

[0056] Comparative Example 3

[0057] The only difference from Example 1 is that the stabilizer is a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 0.2:1:1, and the other components and concentrations are the same as those in Example 1.

[0058] Comparative Example 4

[0059] The only difference from Example 1 is that the surfactant A is only Tween 80, and the other components and concentrations are the same as those in Example 1.

[0060] Comparative Example 5

[0061] The only difference from Example 1 is that the surfactant A is a mixture of Tween 80 and PVP K30 in a mass ratio of 1:3, and the other components and concentrations are the same as those in Example 1.

[0062] Comparative Example 6

[0063] The only difference from Example 1 is that the surfactant B is only Triton X-100, and the other components and concentrations are the same as those in Example 1.

[0064] Comparative Example 7

[0065] The only difference from Example 1 is that surfactant B is a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:1, and other components and concentrations are the same as those in Example 1.

[0066] Comparative Example 8

[0067] A procalcitonin assay kit comprises a mycoavidin-coated photosensitive microsphere, and R1 reagent and R2 reagent which are independent of each other.

[0068] The R1 reagent consists of the following components:

[0069] 50 mg / L of luminescent microparticles coated with procalcitonin monoclonal antibody, 4 g / L of surfactant A (a mixture of Tween 80 and PVP K30 in a mass ratio of 1:1), 0.2 g / L of stabilizer (a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 3:2:1), and 2 g / L of glycine buffer (pH = 6.5).

[0070] R2 reagent consists of the following components:

[0071] Biotin-labeled procalcitonin monoclonal antibody 5 mg / L, surfactant B 5 ml / L (a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:3), sodium chloride 6 g / L, sodium azide 1 mL / L.

[0072] Comparative Example 9

[0073] A procalcitonin chemiluminescent immunoassay kit was prepared according to Example 1 of Chinese invention patent CN 112904023A.

[0074] When the kit of the present invention is used, its determination method is to adopt Zhejiang Quark fully automatic chemiluminescence immunoassay instrument, use rate method to measure, the detection main wavelength is 680nm, and the operation is as follows: add 15μL of physiological saline, sample or standard, then add 25μL of R1 reagent and 25μL of R2 reagent, incubate for 6 minutes for the first time, and read the absorbance A1; finally, add 100μL of streptavidin-coated photosensitive microspheres, incubate for 6 minutes for the second time, read the absorbance A2 after 5 minutes, and calculate ΔA=(A2-A1).

[0075] Procalcitonin content (ng / mL) = (ΔA sample ÷ ΔA calibrator) × calibrator concentration.

[0076] Sample requirements:

[0077] (1) Serum: Serum samples must be collected using standard test tubes to ensure that the samples have been fully agglutinated before centrifugation. Serum samples can be stored at 2-8°C, and it is recommended that the test be completed within 24 hours after sampling. For long-term storage, it is recommended to store at -20°C for three months after aliquoting to avoid repeated freezing and thawing.

[0078] (2) Plasma: Both heparin plasma and EDTA plasma are suitable. Plasma samples can be stored at 2-8°C, and it is recommended to complete the test within 24 hours after sampling. If stored for a long time, it is recommended to store at -20°C for three months after aliquoting to avoid repeated freezing and thawing.

[0079] (3) Whole blood: Whole blood sample with EDTA anticoagulant added. The sample needs to be turned upside down several times before testing. It can be stored at 2-8°C. It is recommended to complete the test within 24 hours after sampling. If stored for a long time, it is recommended to store it at -20°C for three months after aliquoting to avoid repeated freezing and thawing.

[0080] (4) Samples with severe hemolysis, lipemia or turbidity cannot be used for measurement.

[0081] (5) Before testing, the sample and test solution must be equilibrated to ambient temperature before use.

[0082] Experiment 1: Precision Test

[0083] The kits described in Examples 1-3 and Comparative Examples 1-9 were used to test samples of two different concentrations, 0.2 ng / mL and 20 ng / mL. Each concentration was tested 10 times, and the mean value, standard deviation (SD) and coefficient of variation (CV) of the precision of the kit were calculated. The coefficient of variation is usually used to measure the precision of a determination method. The smaller the coefficient of variation, the better the precision of the result of the determination method. For clinical chemistry test items, the method precision with a coefficient of variation of less than 5% is generally recognized to be acceptable. The results are shown in Table 1.

[0084] Coefficient of variation (%) = (standard deviation / mean value) × 100%.

[0085] Table 1 Precision test results

[0086]

[0087] Experiment 2: Stability test

[0088] The kits provided in Examples 1-3 and Comparative Examples 1-9, each containing 12 groups in parallel, were stored together in a refrigerator at 2-8°C. Under the storage condition of 2-8°C, the same standard sample with a procalcitonin antigen content of 10 mg / L was measured at 0 month, 6 months and 12 months, respectively. Each sample was measured 10 times and the average was taken. The results are shown in Table 2.

[0089] Table 2 Stability test results

[0090]

[0091]

[0092] As shown in Table 2, the kits described in Examples 1-3 provided by the present invention have basically no change within 12 months under the condition of 2-8°C, and have good stability; while the reagents in the kits described in Comparative Examples 1-9 have obvious changes within 12 months. The stability of the kit of Example 1 is better than that of the kits of Comparative Examples 1-9, indicating that the present invention significantly improves the stability of the procalcitonin assay kit under the combined action of the stabilizer, surfactant A, surfactant B, and the addition of multiple solvents.

[0093] Experiment 3: Accuracy Test

[0094] The test kits described in Examples 1-3 and Comparative Examples 1-9 were used to measure samples with a test concentration of 10 mg / L, repeated 5 times, and the average was taken to calculate the relative deviation (CB). The test results are shown in Table 3. For clinical chemistry test items, the relative deviation does not exceed ±15%, which is considered to have excellent accuracy.

[0095] Table 3 Accuracy test results

[0096]

[0097]

[0098] The relative deviations in Table 3 all do not exceed ±0.60%, indicating that the kit provided by the present invention has high accuracy.

[0099] Experiment 4: Specificity test

[0100] Humankatacalcin, humancalcitonin, humanalpha CGRPb and humanbeta CGRP, which have similar structures to procalcitonin, were selected and prepared into samples with concentrations greater than physiological concentrations. The present method was used for determination and the cross-reaction rate was calculated. The results are shown in Table 4.

[0101] Table 4 Specificity test results

[0102]

[0103] As shown in Table 4, the cross-reaction rates of the present method with procalcitonin, calcitonin, calcitonin gene-related peptide b and calcitonin gene-related peptide were all less than 0.01%, indicating that the procalcitonin assay kit of the present invention has good specificity.

[0104] Experiment 5: Linearity Test

[0105] A pure procalcitonin sample was taken at 500 ng / mL and diluted to prepare 8 samples of different concentrations, namely 200 ng / mL, 150 ng / mL, 100 ng / mL, 50 ng / mL, 10 ng / mL, 5 ng / mL, 2.5 ng / mL and 0 ng / mL. The kit described in Example 1 was used to detect 8 concentration gradients of standard products. Each gradient concentration was detected 3 times and the average value was taken. The average value of the measured value and the theoretical concentration of the standard product were subjected to regression analysis. The X-axis represents the theoretical concentration, the Y-axis represents the average of the measured value, and the general R 2 ≥0.9900 is considered to have good linearity, see the results Figure 1 .

[0106] The obtained linear regression equation is: y = 0.9984x-0.1934, correlation coefficient: R 2 =0.9999. The results show that the kit provided by the present invention has a good linear correlation.

[0107] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A procalcitonin assay kit, characterized in that: The procalcitonin assay kit comprises R1 reagent, R2 reagent and streptavidin-coated photosensitive microspheres which are independent of each other, wherein the R1 reagent comprises 30-40 mg / L of luminescent microparticles coated with procalcitonin monoclonal antibody, 2-3 g / L of surfactant A and 0.5-1 g / L of stabilizer, and the R2 reagent comprises 6-10 mg / L of biotin-labeled procalcitonin monoclonal antibody and 2-4 mL / L of surfactant B. The surfactant A is a mixture of Tween 80 and PVP K30 in a mass ratio of 1-2:1, the surfactant B is a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:2-4, and the stabilizer is a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 2-10:1-4:

1.

2. The procalcitonin assay kit according to claim 1, characterized in that The surfactant A is a mixture of Tween 80 and PVP K30 in a mass ratio of 1-1.5:1, and the surfactant B is a mixture of Triton X-100 and polyoxyethylene alkyl ether EMULGEN 709 in a mass ratio of 1:2-3.

3. The procalcitonin assay kit according to claim 1, characterized in that The stabilizer is a mixture of bovine serum albumin, gelatin and tetrasodium ethylenediaminetetraacetic acid in a mass ratio of 2-5:1-3:

1.

4. The procalcitonin assay kit according to any one of claims 1 to 3, characterized in that The R1 reagent includes a buffer, and the R2 reagent includes an osmotic agent and a preservative.

5. The procalcitonin assay kit according to claim 4, characterized in that The buffer is selected from one or more of a phosphate buffer, a Tris buffer, a MOPS buffer, a PBS buffer, a MES buffer and a glycine buffer, and the pH of the buffer is 6.5-7.

5.

6. The procalcitonin assay kit according to claim 4, characterized in that: The osmotic agent is selected from one or more of sodium chloride, β-cyclodextrin and carboxymethyl cellulose.

7. The procalcitonin assay kit according to claim 4, characterized in that: The preservative is selected from one or more of phenol, sodium benzoate and sodium azide.

8. The procalcitonin assay kit according to claim 4, characterized in that: The R1 reagent includes 4-6 g / L of buffer, and the R2 reagent includes 4-6 g / L of osmotic agent and 1-1.5 mL / L of preservative.

9. Use of the procalcitonin assay kit according to any one of claims 1 to 8 in the assay of procalcitonin in serum for non-diagnostic purposes.

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