A calibrator buffer, calibrator, and kit for procalcitonin detection.

Abstract

This application belongs to the field of diagnostic reagent technology, and particularly relates to a calibrator buffer solution, calibrator, and reagent kit for procalcitonin detection kit. The calibrator buffer solution for procalcitonin detection kit provided by this application has good stability for preserving procalcitonin antigen stock solution, while avoiding the lyophilization process, reducing the manufacturer's process flow and costs, and improving the user experience by eliminating the need for reconstitution. As a buffer solution specifically for procalcitonin calibrators, it can solve the technical problem in the prior art where the need for reconstitution of lyophilized powder calibrators leads to excessive detection errors.

Description

Technical Field

[0001] This application belongs to the field of diagnostic reagent technology, and in particular relates to a calibrator buffer, calibrator and reagent kit for procalcitonin detection kit. Background Technology

[0002] Procalcitonin (PCT) is a hormone precursor composed of 116 amino acids with a molecular weight of approximately 12.7 kDa. PCT is expressed by neuroendocrine cells (including C cells in thyroid, lung, and pancreatic tissues) and is enzymatically broken down into (immature) calcitonin, carboxyl-terminal peptide, and amino-terminal peptide.

[0003] Healthy individuals typically have only a small amount of procalcitonin (PCT) in their blood, usually around 0.05 ng / mL. Bacterial infections have shown that PCT levels significantly increase after infection. Animal models have demonstrated that PCT can be expressed in various tissues during sepsis. PCT in sepsis patients contains only 114 amino acids, lacking the N-terminal Ala-Pro. Elevated PCT levels are seen in bacterial sepsis, especially severe sepsis and septic shock. PCT can serve as a prognostic indicator for sepsis patients and is also a reliable indicator of acute severe pancreatitis and its major complications. For patients with community-acquired respiratory infections and air conditioning-induced pneumonia, PCT can be used as an antibiotic. The selection and efficacy assessment of indicators; currently, the in vitro quantitative detection of procalcitonin (PCT) mainly relies on diagnostic kits. PCT diagnostic kits are used in conjunction with PCT calibrators. For example, Roche Diagnostics provides calibrators for its PCT diagnostic kits. However, these calibrators are lyophilized powders. When using these lyophilized powder calibrators, a certain volume of purified water needs to be manually added beforehand for reconstitution. Some manufacturers even require transferring the calibrators to other suitable containers after reconstitution. This manual operation not only introduces errors but also leads to batch-to-batch variations, resulting in a poor user experience. The manufacturer's costs also increase due to the lyophilization process. Furthermore, the biochemical properties and concentrations of different diagnostic markers vary greatly in the human body, making it difficult to use calibrators interchangeably. Summary of the Invention

[0004] In view of this, this application provides a calibrator buffer, calibrator, and kit for a procalcitonin detection kit, which solves the technical problem that the lyophilized powder calibrator in the prior art needs to be reconstituted, resulting in excessive detection error.

[0005] The first aspect of this application provides a calibrator buffer for a procalcitonin assay kit, comprising salt, surfactant, preservative, protein stabilizer, and buffer solution.

[0006] Preferably, the protein stabilizer includes Xibao Biotechnology protein stabilizer DCL0009A, bovine serum albumin, and gelatin.

[0007] Preferably, the salt is sodium chloride or potassium chloride;

[0008] Preferably, the surfactant is at least one of polyoxyethylene sorbitan fatty acid ester, polyoxyethylene-8-octylphenyl ether, and lauryl alcohol polyoxyethylene ether;

[0009] Preferably, the preservative is at least one selected from ProClin, sodium azide, penicillin, streptomycin, gentamicin sulfate, and 5-bromo-5-nitro-1,3-dioxanepropanediol;

[0010] Preferably, the protein stabilizer further includes at least one of human serum albumin, casein, and fish gelatin;

[0011] Preferably, the buffer solution is at least one of PBS, TRIS, MES, HEPES, or MOPS buffer solutions.

[0012] Preferably, the salt is sodium chloride;

[0013] The surfactant is polyoxyethylene-8-octylphenyl ether;

[0014] The preservative is ProClin;

[0015] The buffer solution is a TRIS buffer solution.

[0016] Preferably, the surfactant polyoxyethylene sorbitan fatty acid ester is from the TW20, TW40 or TW80 series, the surfactant oxyethylene-8-octylphenyl ether is from the TX45, TX100, TX305 or TX405 series, and the surfactant lauryl ether is from the Brij23, Brij30, Brij35, Brij58 or Brij97 series.

[0017] The preservative ProClin is either ProClin 300 or ProClin 950 series.

[0018] Preferably, the calibrator buffer solution of the procalcitonin assay kit comprises the following proportions;

[0019]

[0020] The buffer solution has a concentration of 0.01M to 0.1M and a pH of 5.0 to 9.0.

[0021] Preferably, the calibrator buffer solution of the procalcitonin assay kit comprises the following proportions;

[0022]

[0023] The TRIS buffer solution had a concentration of 0.02 M and a pH of 8.

[0024] A second aspect of this application provides a calibrator for a procalcitonin detection kit, comprising procalcitonin and a buffer solution for the procalcitonin detection kit calibrator.

[0025] Preferably, the calibrators for the procalcitonin assay kit include a first calibrator for the procalcitonin assay kit and a second calibrator for the procalcitonin assay kit.

[0026] The concentration of the calibrator for the first procalcitonin assay kit is 0.48–0.52 ng / mL;

[0027] The concentration of the calibrator for the second procalcitonin assay kit is 9.60–10.2 ng / mL.

[0028] A third aspect of this application provides a procalcitonin detection kit, including a procalcitonin detection reagent, a procalcitonin detection quality control product, and a calibrator for the procalcitonin detection kit.

[0029] It should be noted that since the calibration buffer provided in this application does not require manual reconstitution of the lyophilized powder, it is beneficial to the accuracy of the test results after long-term storage of the procalcitonin test kit when used with the procalcitonin test reagent and the procalcitonin test quality control product.

[0030] In summary, this application provides a calibrator buffer solution, calibrator, and reagent kit for procalcitonin assay kits. The calibrator buffer solution, comprising salt, surfactant, preservative, protein stabilizer, and buffer solution, maintains the signal retention rate of the procalcitonin assay kit calibrator for more than 12 months at low temperatures (2–8°C) and for more than 7 days at 37°C. This demonstrates that the calibrator buffer solution provided in this application can maintain the stability of the procalcitonin antigen stock solution over a long period. Furthermore, as a novel preservation process for procalcitonin calibrators, it avoids the drawbacks of conventional procalcitonin calibrators, which require lyophilization to obtain lyophilized powder. The reconstitution of lyophilized powder can lead to uncontrollable factors and reduced accuracy of clinical test results, thus solving the technical problem of excessive detection errors caused by the reconstitution of lyophilized powder calibrators in existing technologies. Detailed Implementation

[0031] This application provides a calibrator buffer solution, calibrator, and reagent kit for procalcitonin detection, which solves the technical problem in the prior art where the lyophilized powder calibrator needs to be reconstituted, resulting in excessive detection error.

[0032] The technical solutions of this application will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0033] Example 1

[0034] Given that existing procalcitonin diagnostic kits use lyophilized powder as the calibrator, which requires manual reconstitution with purified water, this manual process can easily lead to detection errors. Example 1 of this application provides a novel procalcitonin calibrator preservation process. This process preserves the procalcitonin calibrator using a calibrator buffer solution. The buffer solution for the procalcitonin test kit comprises salts, surfactants, preservatives, protein stabilizers, and a buffer solution. This buffer solution provides good long-term stability for procalcitonin, ensuring the calibrator remains stable for 12 months at 2–8°C. This avoids the need for reconstitution of lyophilized powder prepared using conventional preservation methods, thus improving the accuracy of clinical test results. This solves the technical problem of excessive detection errors caused by the need for reconstitution of lyophilized powder calibrators in existing technologies. Furthermore, it reduces the lyophilization process, lowers the manufacturer's process flow and costs, and eliminates the need for reconstitution, improving the user experience.

[0035] For the specific types of salts, surfactants, preservatives, protein stabilizers, and buffer solutions used in the calibrator buffer solution for the procalcitonin diagnostic kit, this application preferably uses sodium chloride or potassium chloride as the salt; the surfactant is at least one of polyoxyethylene sorbitan fatty acid ester, polyoxyethylene-8-octylphenyl ether, and lauryl polyoxyethylene ether; the preservative is at least one of ProClin, sodium azide, penicillin, streptomycin, gentamicin sulfate, and 5-bromo-5-nitro-1,3-dioxanepropanediol; the protein stabilizer is at least one of Xibao Biotechnology protein stabilizer DCL0009A, bovine serum albumin, human serum albumin, casein, gelatin, and fish gelatin; and the buffer solution is at least one of PBS, TRIS, MES, HEPES, or MOPS buffer solution.

[0036] This application provides 29 formulations for the calibrator buffer solution used with the procalcitonin diagnostic kit, as shown in Table 1.

[0037]

[0038] In Table 1, A1–A5 are selected from PBS, TRIS, MES, HEPES, and MOPS buffer solutions, respectively; B1–B6 are selected from Xibao Biotechnology protein stabilizer (product number: DCL0009A), bovine serum albumin, human serum albumin, casein, gelatin, and fish gelatin protein stabilizer, respectively; C1–C2 are selected from sodium chloride and potassium chloride, respectively; D1–D12 are selected from TW20, TW40, TW80, TX45, TX100, TX305, TX405, Brij23, Brij30, Brij35, Brij58, and Brij97, respectively; E1–E7 are selected from PC300, PC950, sodium azide, penicillin, streptomycin, gentamicin sulfate, and 5-bromo-5-nitro-1,3-dioxanepropanediol, respectively.

[0039] For the calcitonin concentration in the calibrator buffer solution of the calcitonin diagnostic kit provided in this application, the preferred composition is 0.1–4.0 wt% salt, 0.01–3.00 wt% surfactant, 0.05%–1 wt% preservative, 0.01–20.00 wt% protein stabilizer, with the remainder being buffer solution; wherein the buffer solution concentration is 0.01 M–0.05 M, pH is 5.0–9.0, and the total amount of salt, surfactant, preservative, protein stabilizer, and buffer solution is 100 wt%.

[0040] For the optimal formulation of the calibrator buffer solution for the procalcitonin diagnostic kit provided in this application, the preferred formulation includes 0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w% ProClin, 2 w% Xibao Biotechnology protein stabilizer DCL0009A, 1 w% bovine serum albumin, 1 w% gelatin, and the balance being TRIS buffer solution; wherein the buffer solution concentration is 0.02M, pH is 8, and the total amount of sodium chloride salt, polyoxyethylene-8-octylphenyl ether surfactant, ProClin preservative, Xibao Biotechnology protein stabilizer DCL0009A, bovine serum albumin, gelatin protein stabilizer, and TRIS buffer solution is 100 w.

[0041] Example 2

[0042] Example 2 of this application provides a method for preparing calcitonin test kit calibrators using the calibrator buffer solution described in Example 1, to evaluate whether the buffer solution provided in this application will cause calibrator errors. The preparation method involves mixing the salt, surfactant, preservative, protein stabilizer, and buffer solution formulation described in Example 1 with the calcitonin antigen stock solution. The calcitonin test kit calibrators are low-concentration (Cal1) and high-concentration calcitonin test kit calibrators (Cal2). The prepared low-concentration (Cal1) and high-concentration calcitonin test kit calibrators (Cal2) are detected using a fully automated chemiluminescence immunoassay analyzer (iNova3000). The calcitonin luminescence values ​​are shown in Tables 2-5.

[0043]

[0044]

[0045] Table 2

[0046]

[0047] Table 3

[0048]

[0049]

[0050] Table 4

[0051] Source of mutation SS df MS F <![CDATA[F 0.05 ]]> p Total variation 8.36053099 49 Bottle mutation 1.76251419 9 0.195835 1.187 2.124 >0.05 In-bottle mutation 6.5980168 40 0.16495

[0052] Table 5

[0053] As shown in Tables 2-5, the 10 groups of low-concentration procalcitonin assay calibrators (Cal1) prepared by uniformly mixing the buffer solution provided in Example 1 of this application with the procalcitonin antigen stock solution exhibited small variability, and the variability among multiple tests of any one group of low-concentration procalcitonin assay calibrators (Cal1) was also small. This indicates that the buffer solution provided in this application can be used with low-concentration (0.48–0.52 ng / mL) procalcitonin assay calibrators (Cal1) with small preparation error with the procalcitonin antigen stock solution. Simultaneously, the high-concentration procalcitonin assay calibrator (Cal2) also showed similar results, indicating that the buffer solution provided in this application can also be used with high-concentration (9.60–10.2 ng / mL) procalcitonin assay calibrators (Cal2) with small preparation error with the procalcitonin antigen stock solution. Therefore, it can be used to preserve procalcitonin and has the potential to be used as a buffer solution for procalcitonin calibrators. The formulations for bottles 1-10 are as follows:

[0054] Bottle number 1: 0.02M-TRIS (pH8.0)+1w%BSA+0.1w%PC300+0.9w%NaCl+0.1w%TX100;

[0055] Bottle No. 2: 0.1M PBS (pH 7.4) + 1 w% BSA + 0.1 w% PC300 + 0.1 w% TX100;

[0056] Bottle number 3: 0.05M-TRIS(pH8.0)+2w%BSA+0.1w%PC300+0.9w%NaCl+0.1w%TX100+2w%DCL0009A+0.05%TW20;

[0057] Bottle No. 4: 0.05M-TRIS (pH 8.0) + 2w% BSA + 0.1w% PC300 + 0.9w% NaCl + 0.1w% TX100 + 2w% gelatin;

[0058] Bottle No. 5: 0.02M-TRIS(pH8.0)+1w%BSA+0.1w%PC300+0.9w%NaCl+0.1w%TX100+2w%DCL0009A;

[0059] Bottle No. 6: 0.05M-TRIS (pH 8.0) + 2w% BSA + 0.1w% PC300 + 0.9w% NaCl + 0.1w% TX100 + 2w% gelatin + 0.1w% gentamicin sulfate;

[0060] Bottle No. 7: 0.02M-TRIS(pH8.0)+1w%BSA+0.1w%PC300+0.9w%KCl+0.1w%TX100+2w%DCL0009A;

[0061] Bottle No. 8: 0.02M-TRIS (pH 8.0) + 1w% BSA + 0.1w% PC300 + 0.9w% NaCl + 0.1w% Brij35;

[0062] Bottle number 9: 0.05M-HEPES (pH7.4)+2w%BSA+0.1w%PC300+0.9w%NaCl+0.1w%Brij35;

[0063] Bottle No. 10: 0.02M-TRIS (pH 8.0) + 2w% BSA + 0.1w% Sodium azide + 0.9w% KCl + 0.1w% TX100 + 2w% DCL0009A + 2w% Gelatin.

[0064] Experimental Example 1

[0065] Example 1 of this application provides a buffer solution for preparing calibrators for the procalcitonin assay kit to optimize the effect of the buffer formulation provided in this application in preserving procalcitonin. The preparation method is as follows: 1 w% BSA and 0.05 w% PC300, as well as different buffer solutions, are mixed evenly with the procalcitonin antigen stock solution to prepare calibrators for the procalcitonin assay kit, including low concentration (Cal1, 0.48-0.52 ng / mL) and high concentration (Cal2, 9.60-10.2 ng / mL). The prepared low concentration (Cal1) and high concentration (Cal2) calibrators were stored at 37°C. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviation was calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results of the procalcitonin luminescence values ​​are shown in Table 6.

[0066]

[0067]

[0068] Table 6

[0069] As shown in Table 6, among the buffer solutions provided in this application, the TRIS buffer exhibits the best preservation effect on the procalcitonin antigen stock solution, with a high signal retention rate compared to other buffer solutions. Furthermore, this application has optimized the optimal concentration and pH value of the TRIS buffer. The preparation method involves uniformly mixing 1 w% BSA and 0.05 w% PC300, along with different concentrations or pH values ​​of TRIS buffer solutions, with the procalcitonin antigen stock solution to prepare the procalcitonin detection kit calibrator, including low concentrations (Cal1, 0.48–0.52 ng / mL) and high concentrations. The calibrators for the high-concentration procalcitonin assay kit (Cal2, 9.60–10.2 ng / mL) were prepared and stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the procalcitonin assay kit (magnetic microparticle chemiluminescence method). The luminescence value results of the procalcitonin assay kit are shown in Tables 7 and 8.

[0070]

[0071]

[0072] Table 7

[0073]

[0074] Table 8

[0075] As can be seen from Table 7-8, when the TRIS buffer solution concentration is 0.02M and the pH is 8, it has the best preservation effect on the procalcitonin antigen stock solution, and the signal retention rate is higher than that of other buffer solutions with different concentrations and pH values.

[0076] Experiment Example 2

[0077] Example 2 of this application provides a buffer solution for preparing calibrators for the procalcitonin assay kit to optimize the effect of the buffer formulation provided in this application in preserving calcitonin. The preparation method is as follows: 1 w% BSA, 0.05 w% PC300, and different protein stabilizers are mixed evenly with the calcitonin antigen stock solution to prepare calibrators for the procalcitonin assay kit, including low concentration (Cal1, 0.48-0.52 ng / mL) and high concentration calibrators (Cal2, 9.60-10.2 ng / mL). The prepared low concentration (Cal1) and high concentration calibrators (Cal2) were stored at 37°C. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviation was calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the calcitonin assay kit (magnetic microparticle chemiluminescence method). The results of the detected calcitonin luminescence values ​​are shown in Table 9.

[0078]

[0079] Table 9

[0080] As shown in Table 9, among the buffer solutions provided in this application, the Xibao Biotechnology protein stabilizer DCL0009A, bovine serum albumin, and gelatin exhibit the best preservation effect on the calcitonin antigen stock solution, with high signal retention rates compared to other protein stabilizers. Furthermore, this application has optimized the optimal concentrations of Xibao Biotechnology protein stabilizer DCL0009A, bovine serum albumin, and gelatin. The preparation method involves uniformly mixing 1 w% BSA and 0.05 w% PC300, along with different concentrations of Xibao Biotechnology protein stabilizer DCL0009A, bovine serum albumin, and gelatin, with the calcitonin antigen stock solution to prepare the calcitonin detection kit calibrator. The calibrators included low-concentration (Cal1, 0.48–0.52 ng / mL) and high-concentration calcitonin assay kit calibrators (Cal2, 9.60–10.2 ng / mL). After preparation, the low-concentration (Cal1) and high-concentration calcitonin assay kit calibrators (Cal2) were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the calcitonin assay kit (magnetic microparticle chemiluminescence method). The calcitonin luminescence value results are shown in Table 10-12.

[0081]

[0082] Table 10

[0083]

[0084]

[0085] Table 11

[0086]

[0087] Table 12

[0088] As can be seen from Table 10-12, when the concentrations of Xibao Biotech protein stabilizer DCL0009A, bovine serum albumin, and gelatin are 2w%, 2w% (including 1w% BSA base solution), and 1w%, respectively, the preservation effect on calcitonin antigen stock solution is the best, and the signal retention rate is higher than that of buffer solutions containing other concentrations of Xibao Biotech protein stabilizer DCL0009A, bovine serum albumin, and gelatin.

[0089] Experimental Example 3

[0090] Example 3 of this application provides a buffer solution for preparing calibrators for the procalcitonin assay kit to optimize the effect of the buffer formulation provided in this application in preserving procalcitonin. The preparation method is as follows: 1 w% BSA and 0.05 w% PC300, along with different salts, are mixed evenly with the procalcitonin antigen stock solution to prepare calibrators for the procalcitonin assay kit, including low concentration (Cal1, 0.48–0.52 ng / mL) and high concentration (Cal2, 9.60–10.2 ng / mL). The prepared low concentration (Cal1) and high concentration (Cal2) calibrators were stored at 37°C. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using a fully automated chemiluminescence immunoassay analyzer (iNova3000) from Yangpu Medical and a procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results of the detected procalcitonin luminescence values ​​are shown in Table 13.

[0091]

[0092] Table 13

[0093] As can be seen from Table 13, when the salt is selected from sodium chloride and the concentration is 0.9 w%, the preservation effect of procalcitonin antigen stock solution is the best, and the signal retention rate is higher than that of buffer solutions with other salts.

[0094] Experiment Example 4

[0095] Example 4 of this application provides a buffer solution for preparing calibrators for the procalcitonin assay kit to optimize the effect of the buffer formulation provided in this application in preserving procalcitonin. The preparation method is as follows: 1 w% BSA and 0.05 w% PC300, along with different surfactants, are mixed evenly with the procalcitonin antigen stock solution to prepare calibrators for the procalcitonin assay kit, including low concentration (Cal1, 0.48–0.52 ng / mL) and high concentration (Cal2, 9.60–10.2 ng / mL). The prepared low concentration (Cal1) and high concentration (Cal2) calibrators were stored at 37°C. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using a fully automated chemiluminescence immunoassay analyzer (iNova3000) from Yangpu Medical and a procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results of the detected procalcitonin luminescence values ​​are shown in Table 14.

[0096]

[0097]

[0098] Table 14

[0099] As shown in Table 14, among the buffer solutions provided in this application, surfactant TX100 exhibits the best preservation effect on the procalcitonin antigen stock solution, with a higher signal retention rate than other surfactants. Furthermore, this application has optimized the optimal concentration of surfactant TX100. The preparation method involves uniformly mixing 1 w% BSA, 0.05 w% PC300, and different concentrations of surfactant TX100 with the procalcitonin antigen stock solution to prepare the procalcitonin detection kit calibrator, including low concentrations (Cal1, 0.48–0.52 ng / mL). The high-concentration procalcitonin assay kit calibrator (Cal2, 9.60–10.2 ng / mL) and the prepared low-concentration (Cal1) and high-concentration procalcitonin assay kit calibrators (Cal2) were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results of the procalcitonin luminescence values ​​are shown in Table 15.

[0100]

[0101]

[0102] Table 15

[0103] As can be seen from Table 15, when the concentration of surfactant TX100 is 0.1 w%, it has the best preservation effect on procalcitonin antigen stock solution, and the signal retention rate is higher than that of surfactant TX100 at other concentrations.

[0104] Experimental Example 5

[0105] Example 5 of this application provides a buffer solution for preparing calibrators for the procalcitonin assay kit to optimize the effect of the buffer formulation provided in this application on preserving procalcitonin. The preparation method is as follows: 1 w% BSA, 0.05 w% PC300, and different preservatives are mixed evenly with the procalcitonin antigen stock solution to prepare calibrators for the procalcitonin assay kit, including low concentration (Cal1, 0.48-0.52 ng / mL) and high concentration (Cal2, 9.60-10.2 ng / mL). The prepared low concentration (Cal1) and high concentration (Cal2) calibrators were stored at 37°C. Solutions containing the target protein were taken on day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviation was calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results of the detected procalcitonin luminescence values ​​are shown in Table 16.

[0106]

[0107]

[0108] Table 16

[0109] As shown in Table 16, among the buffer solutions provided in this application, the preservative ProClin300 exhibits the best preservation effect on the procalcitonin antigen stock solution, with a higher signal retention rate than other preservatives. Furthermore, this application has optimized the optimal concentration of the preservative ProClin300. The preparation method involves uniformly mixing 1 w% BSA, 0.05 w% PC300, and different concentrations of the preservative ProClin300 with the procalcitonin antigen stock solution to prepare the procalcitonin detection kit calibrator, including low concentrations (Cal1, 0.48–0.52 ng / L). The low-concentration (Cal1) and high-concentration procalcitonin assay kit calibrators (Cal2, 9.60–10.2 ng / mL) were prepared and stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The luminescence values ​​of the calibrators were measured and the deviations were calculated using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results of the procalcitonin luminescence values ​​are shown in Table 17.

[0110]

[0111]

[0112] Table 17

[0113] As can be seen from Table 17, when the concentration of the preservative ProClin300 is 0.1 w% (including 0.05 w% ProClin300 base solution), it has the best preservation effect on the procalcitonin antigen stock solution and the signal retention rate is higher than that of other concentrations of preservative ProClin300 buffer solution.

[0114] Experimental Example 6

[0115] Example 6 of this application provides a performance test of the procalcitonin assay kit calibrator prepared by uniformly mixing the procalcitonin antigen stock solution with the calibrator buffer solution, and the commercially available lyophilized powder calibrator provided by Roche Diagnostics, to compare their effects on preserving procalcitonin (37°C). The test method includes reconstituted the lyophilized powder calibrator provided by Roche Diagnostics with the procalcitonin assay kit calibrator, and then preparing a solution containing 0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w% ProClin, 2 w% Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 1 w% gelatin, with the remainder being TRIS buffer solution; wherein the concentration of the TRIS buffer solution is 0.0. The calibrators for the procalcitonin assay kit were prepared by mixing 25M, pH 8.0, and the procalcitonin antigen stock solution. The prepared calibrators included low-concentration (Cal1, 0.48–0.52 ng / mL) and high-concentration (Cal2, 9.60–10.2 ng / mL) calibrators. The prepared low-concentration (Cal1) and high-concentration (Cal2) calibrators were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The results were obtained using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results are shown in Table 18.

[0116]

[0117] Table 18

[0118] As shown in Table 18, the lyophilized powder calibrators provided by Roche Diagnostics exhibit poor stability after reconstitution. After 7 days, the signal retention value of the low-concentration (Cal1, 0.48–0.52 ng / mL) calibrator for the procalcitonin assay kit was only 60%, while the signal retention value of the high-concentration (Cal2, 9.60–10.2 ng / mL) calibrator was 51%. In contrast, the calibrator buffer solution provided in this application (0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w% ProClin, 2 w% Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 1 w% gelatin, with the remainder being TR) is significantly better. The TRIS buffer solution (0.025M, pH 8.0) showed a signal retention rate of 94% for low-concentration (Cal1, 0.48–0.52 ng / mL) procalcitonin assay kit calibrators after 7 days, while the signal retention rate for high-concentration (Cal2, 9.60–10.2 ng / mL) procalcitonin assay kit calibrators was 99%. This demonstrates that the procalcitonin assay kit calibrator buffer solution provided in this application can maintain the stability of the procalcitonin antigen stock solution over a long period. It also avoids the need for conventional procalcitonin calibrators to be prepared as lyophilized powder, which increases the manufacturer's production process and costs, and reduces the user experience.

[0119] Example 7

[0120] Example 7 of this application provides a performance test of the procalcitonin assay kit calibrator prepared by uniformly mixing the procalcitonin antigen stock solution with the calibrator buffer solution and the commercially available lyophilized powder calibrator provided by Roche Diagnostics, to compare their effects on preserving procalcitonin (2-8°C). The test method includes reconstituted the lyophilized powder calibrator provided by Roche Diagnostics with the procalcitonin assay kit calibrator, and then preparing a solution containing 0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w% ProClin, 2 w% Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 1 w% gelatin, with the remainder being a TRIS buffer solution; wherein the concentration of the TRIS buffer solution is 0. The calibrators for the procalcitonin assay kit were prepared by mixing 0.25M, pH 8.0, and the procalcitonin antigen stock solution. The prepared calibrators included low-concentration (Cal1, 0.48–0.52 ng / mL) and high-concentration (Cal2, 9.60–10.2 ng / mL) calibrators. The prepared low-concentration (Cal1) and high-concentration (Cal2) calibrators were stored at 37°C. Solutions containing the target protein were taken from day 0 and day 7, respectively. The results were obtained using Yangpu Medical's fully automated chemiluminescence immunoassay analyzer (iNova3000) and the procalcitonin assay kit (magnetic microparticle chemiluminescence method). The results are shown in Table 19.

[0121]

[0122] Table 19

[0123] As shown in Table 19, the calibrator buffer (0.9 w% sodium chloride, 0.1 w% polyoxyethylene-8-octylphenyl ether, 0.1 w% ProClin, 2 w% Siba Biotech protein stabilizer DCL0009A, 1 w% bovine serum albumin, 1 w% gelatin, with the remainder being TRIS buffer solution; wherein the TRIS buffer solution concentration is 0.02 M, pH 8.0) of the procalcitonin assay kit provided in this application resulted in a low concentration (Cal1, 0.48–0.52 ng / mL) after 7 days. The calibrator of the procalcitonin assay kit has a signal retention value as high as 90%, while the high-concentration procalcitonin assay kit calibrator (Cal2, 9.60~10.2ng / mL) has a signal retention value of 90%. This indicates that the calibrator buffer solution of the procalcitonin assay kit provided in this application can maintain the stability of the procalcitonin antigen stock solution for a long time. At the same time, it avoids the need for conventional procalcitonin calibrators to be prepared by lyophilization to obtain lyophilized powder, which increases the manufacturer's production process and cost, and reduces the user's experience.

[0124] The above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application.

Claims

1. A calibrator for a procalcitonin assay kit, characterized in that, Includes procalcitonin and procalcitonin test kit calibrator buffer, wherein the procalcitonin test kit calibrator buffer is composed of the following proportions; Sodium chloride 0.9% w% Polyoxyethylene-8-octylphenyl ether TX100 0.1w% ProClin300 0.1%w% Xibao Biotechnology Protein Stabilizer DCL0009A 2w% Bovine serum albumin 1w% Gelatin 1w% Remaining TRIS buffer solution; The TRIS buffer solution had a concentration of 0.025 M and a pH of 8.

2. The calibrator for a procalcitonin detection kit according to claim 1, characterized in that, The calibrators for the procalcitonin assay kit include a first calibrator for the procalcitonin assay kit and a second calibrator for the procalcitonin assay kit. The concentration of the calibrator for the first procalcitonin assay kit is 0.48~0.52 ng / mL; The concentration of the calibrator for the second procalcitonin assay kit is 9.60~10.2 ng / mL.

3. A procalcitonin assay kit, characterized in that, It includes procalcitonin detection reagents, procalcitonin detection quality control products, and a calibrator for a procalcitonin detection kit as described in claim 1.

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