阅读说明：本技术 一种多项目炎症标志物质控品及其制备方法 (Multi-item inflammation marker quality control product and preparation method thereof ) 是由 张军 李海燕 宁妍 于 2020-05-25 设计创作，主要内容包括：本发明是关于一种多项目炎症标志物质控品及其制备方法,该方法包括以下步骤：采用血清或血浆作为基质液,将其恢复至室温；将得到的血清或血浆用孔径0.22μm-0.65μm的滤膜过滤,去除杂质得到质控品基质液；在得到的基质液中加入以下炎症标志物抗原：降钙素原、C反应蛋白、血清淀粉样蛋白A、白细胞介素-6、肝素结合蛋白,制成高、中、低三个浓度水平的质控品溶液；将制成的三水平质控溶液中加入防腐剂、稳定剂和保护剂,混合均匀；对获得的三水平质控品进行测试,待浓度达到要求时,真空冷冻干燥、密封,2-8℃保存。本发明只需要进行一次质控操作,即可对所需的多个项目进行同步地质量控制,大大简化了操作流程,具有生产成本低、准确度高、操作简单的优点。(The invention relates to a quality control product of a multinomial inflammation marker and a preparation method thereof, wherein the method comprises the following steps: adopting serum or plasma as matrix liquid, and returning to room temperature; filtering the obtained serum or plasma with a filter membrane with pore size of 0.22-0.65 μm, and removing impurities to obtain quality control substance matrix solution; the following inflammation marker antigens were added to the obtained matrix fluid: procalcitonin, C-reactive protein, serum amyloid A, interleukin-6 and heparin binding protein are prepared into quality control solution with high, medium and low concentration levels; adding a preservative, a stabilizer and a protective agent into the prepared three-level quality control solution, and uniformly mixing; testing the obtained three-level quality control product, and freeze-drying under vacuum when the concentration meets the requirement, sealing, and storing at 2-8 deg.C. The invention can synchronously control the quality of a plurality of required projects by only one-time quality control operation, greatly simplifies the operation flow and has the advantages of low production cost, high accuracy and simple operation.)

1. A preparation method of a multi-item inflammation marker quality control product is characterized by comprising the following steps:

the first step, adopting serum or plasma as a substrate liquid, and returning the substrate liquid to room temperature;

secondly, filtering the serum or the plasma obtained in the first step by using a filter membrane with the aperture of 0.22-0.65 μm, and removing impurities to obtain a quality control product matrix solution;

thirdly, adding the inflammation marker antigen with the following concentration into the quality control substance matrix liquid obtained in the second step: procalcitonin: (0-100) ng/mL, C-reactive protein: (0-200) mg/L, serum amyloid A: (0-200) mg/L, interleukin-6: (0-4000) pg/mL, heparin-binding protein: (0-1000) ng/mL, and preparing quality control solution with high, medium and low concentration levels respectively;

fourthly, respectively adding 0.005-0.1 wt% of preservative, 0.5-5 wt% of stabilizer and 0.5-5 wt% of protective agent into the quality control solutions with the three concentration levels prepared in the third step, and uniformly mixing;

and fifthly, respectively detecting the three-level quality control products obtained in the fourth step, and when the concentration meets the requirement, carrying out vacuum freeze drying, sealing and storing at the temperature of 2-8 ℃.

2. The method of claim 1, wherein in the first step, the serum is selected from one of human serum, newborn bovine serum, calf serum and rabbit serum; the plasma is selected from human plasma; the detection results of the hepatitis B, hepatitis C, AIDS and syphilis marker antibodies in the serum or the plasma are all negative.

3. The method of claim 1, wherein in the third step, the low concentration level of the quality control solution comprises the following inflammation marker antigens: procalcitonin: (0.1-0.5) ng/mL; c-reactive protein: (1-5) mg/L; serum amyloid a: (1-10) mg/L; interleukin-6: (4-10) pg/mL; heparin-binding protein: (1-5) ng/mL.

4. The method of claim 1, wherein in the third step, the medium concentration level of the quality control solution comprises the following inflammation marker antigens: procalcitonin: (0.5-2) ng/mL; c-reactive protein: (10-50) mg/L; serum amyloid a: (10-50) mg/L; interleukin-6: (50-200) pg/mL; heparin-binding protein: (10-30) ng/mL.

5. The method of claim 1, wherein in the third step, the high concentration level of the quality control solution comprises the following inflammation marker antigens: procalcitonin: (5-20) ng/mL; c-reactive protein: (50-200) mg/L; serum amyloid a: (100-300) mg/L; interleukin 6: (500-2000) pg/mL; heparin-binding protein: (30-100) ng/mL.

6. The method of claim 1, wherein in the fourth step, the preservative is at least one selected from proclin-300, KroVin300, sodium azide, thimerosal, and gentamicin.

7. The method of claim 1, wherein in the fourth step, the stabilizer is at least one selected from the group consisting of trehalose, sucrose, lactose, chitosan, and dextran.

8. The method of claim 1, wherein in the fourth step, the protective agent is at least one selected from the group consisting of polyethylene glycol, xanthan gum, gelatin, mannitol, glycerol, and bovine serum albumin.

9. The method for preparing a quality control product of a multinomial inflammation marker according to claim 1, wherein the fifth step specifically comprises: and (3) detecting the three-level quality control product obtained in the fourth step by using a fluorescence immunoassay analyzer, and when the concentration of the three-level quality control product meets the requirement that the deviation of the fixed value quality control detection value does not exceed 10% of the standard value, carrying out vacuum freeze drying, sealing and storing at the temperature of 2-8 ℃.

10. A multi-item inflammation marker quality control prepared by the method of any one of claims 1-9.

Technical Field

The invention relates to a preparation method of an inflammation marker quality control product, in particular to a multi-item inflammation marker quality control product and a preparation method thereof.

Background

Inflammation is a defensive response of the body to external stimuli and can be classified into infectious inflammation caused by infection with bacteria or viruses, and non-infectious inflammation caused by non-infectious factors such as autoimmune system deficiency. Currently, most of the diagnostic treatments for infectious diseases rely on conventional methods such as white blood cell counting, classification, blood sedimentation, enzymatic activity, etc. However, these indicators have limitations, and also cause an increase in the number of leukocytes in non-infected states such as pregnancy, strenuous exercise, intoxication and acute hemorrhage.

Inflammation markers are a series of sensitive substances which are increased by infectious inflammation in blood when the body generates a stress reaction under the influence of infection, trauma or other diseases, such as Procalcitonin (PCT), C-reactive protein (CRP), Serum Amyloid A (SAA), Interleukin-6 (Interleukin-6, IL-6), Heparin-binding protein (HBP), etc., and the detection of which is not influenced by antibacterial drugs, immunosuppressive agents and hormones, and thus, the markers are widely regarded as reliable indicators of inflammation detection.

The methodology and the production process of detecting the inflammation marker by various manufacturers on the market are different at present, so that the adverse effect that the detection results obtained by using the products of different manufacturers are inconsistent can occur; at present, detection kits produced by most manufacturers do not contain quality control products, and the quality control products prepared in clinical laboratories have certain limitations; after the hair is infected, the time for increasing the content of each inflammation marker is different, the detection significance of each item is different from the clinical application, and the possibility of missed diagnosis and misdiagnosis exists in the clinical detection by judging the infection degree only by the content of a single marker, so that the clinical detection usually takes the combined detection of multiple markers as the diagnosis basis, and most of the existing quality control products in the market are single-marker quality control with few types. This means that the laboratory need repeat single sign material accuse when carrying out quality control and detect many times, and the operation is comparatively complicated and fussy, and produces a plurality of single projects and need carry out freeze-drying one by one, and work load is big, and the cost is higher. Therefore, a compound quality control product of multiple inflammation markers with good commercial stability is urgently needed clinically, and is used for indoor quality control and indoor quality evaluation in a laboratory.

Disclosure of Invention

In view of the above, the main objective of the present invention is to provide a stable-performance quality control product with multiple inflammation markers and a preparation method thereof, wherein multiple inflammation markers are compounded into a quality control product, the subsequent detection is simple and convenient, the accuracy is high, and the cost can be reduced to meet the clinical use requirements.

The purpose of the invention and the technical problem to be solved are realized by adopting the following technical scheme.

The preparation method of the multi-item inflammation marker quality control product provided by the invention comprises the following steps:

the first step, adopting serum or plasma as a substrate liquid, and returning the substrate liquid to room temperature;

secondly, filtering the serum or the plasma obtained in the first step by using a filter membrane with the aperture of 0.22-0.65 μm, and removing impurities to obtain a quality control product matrix solution;

thirdly, adding the inflammation marker antigen with the following concentration into the quality control substance matrix liquid obtained in the second step: procalcitonin (PCT): (0-100) ng/mL, C-reactive protein (CRP): (0-200) mg/L, Serum Amyloid A (SAA): (0-200) mg/L, interleukin-6 (IL-6): (0-4000) pg/mL, heparin-binding protein (HBP): (0-1000) ng/mL, and preparing quality control solution with high, medium and low concentration levels respectively;

fourthly, respectively adding 0.005-0.1 wt% of preservative, 0.5-5 wt% of stabilizer and 0.5-5 wt% of protective agent into the quality control solutions with the three concentration levels prepared in the third step, and uniformly mixing;

and fifthly, respectively detecting the three-level quality control products obtained in the fourth step, and when the concentration meets the requirement, carrying out vacuum freeze drying, sealing and storing at the temperature of 2-8 ℃.

The purpose of the invention and the technical problem to be solved are further realized by adopting the following technical scheme.

Preferably, in the aforementioned method for preparing a quality control product of multiple item inflammation markers, in the first step, the serum is selected from one of human serum, newborn bovine serum, calf serum and rabbit serum; the plasma is selected from human plasma; the detection results of the hepatitis B, hepatitis C, AIDS and syphilis marker antibodies in the serum or the plasma are all negative.

Preferably, in the above method for preparing a quality control product of multiple items of inflammation markers, in the third step, the quality control product solution at low concentration level contains the following antigens of inflammation markers: procalcitonin (PCT): (0.1-0.5) ng/mL; c-reactive protein (CRP): (1-5) mg/L; serum amyloid a (saa): (1-10) mg/L; interleukin-6 (IL-6): (4-10) pg/mL; heparin Binding Protein (HBP): (1-5) ng/mL.

Preferably, in the above method for preparing a quality control product of multiple items of inflammation markers, in the third step, the medium-concentration level of the quality control product solution contains the following antigens of inflammation markers: procalcitonin (PCT): (0.5-2) ng/mL; c-reactive protein (CRP): (10-50) mg/L; serum amyloid a (saa): (10-50) mg/L; interleukin-6 (IL-6): (50-200) pg/mL; heparin Binding Protein (HBP): (10-30) ng/mL.

Preferably, in the above method for preparing a quality control product of multiple items of inflammation markers, in the third step, the quality control product solution at a high concentration level contains the following antigens of inflammation markers: procalcitonin (PCT): (5-20) ng/mL; c-reactive protein (CRP): (50-200) mg/L; serum amyloid a (saa): (100-300) mg/L; interleukin-6 (IL-6): (500-2000) pg/mL; heparin Binding Protein (HBP): (30-100) ng/mL.

Preferably, in the aforementioned method for preparing a quality control product of multiple item inflammation markers, in the fourth step, the preservative is at least one selected from proclin-300, KroVin300, sodium azide, thimerosal and gentamicin.

Preferably, in the aforementioned method for preparing a quality control product of a multiple item inflammation marker, the stabilizer in the fourth step is at least one selected from trehalose, sucrose, lactose, chitosan and dextran.

Preferably, in the aforementioned method for preparing a quality control product of a multi-item inflammation marker, in the fourth step, the protective agent is at least one selected from the group consisting of polyethylene glycol, xanthan gum, gelatin, mannitol, glycerol and bovine serum albumin.

Preferably, in the preparation method of the quality control product for multiple items of inflammation markers, the fifth step specifically comprises: and (3) detecting the three-level quality control product obtained in the fourth step by using a fluorescence immunoassay analyzer, and after the concentration of the three-level quality control product meets the requirement that the deviation of the fixed value quality control detection value does not exceed 10% of the standard value, carrying out vacuum freeze drying, sealing and storing at the temperature of 2-8 ℃.

The purpose of the invention and the technical problem to be solved are further realized by adopting the following technical scheme.

According to the invention, the multi-item inflammation marker quality control product is prepared by the method.

Compared with the prior art, the invention has the following beneficial effects:

1. the preparation method of the multi-project inflammation marker quality control product provided by the invention can be used for synchronously controlling the quality of a plurality of required projects only by performing one-time quality control operation, thereby greatly simplifying the operation process and having the advantages of low production cost, high accuracy and simplicity in operation.

2. According to the preparation method of the multi-item inflammation marker quality control product, disclosed by the invention, the multiple inflammation markers are compounded and then are simultaneously detected, so that the interference is reduced, and the quality control operation is simplified.

3. The preparation method of the quality control product of the multi-item inflammation marker reduces the times of freeze drying in the production process and reduces the cost.

4. The quality control product of the multi-item inflammation marker uses serum or plasma as a matrix liquid, has good clinical consistency, and improves the detection accuracy.

The foregoing is a summary of the present invention, and in order to provide a clear understanding of the technical means of the present invention and to be implemented in accordance with the present specification, the following is a detailed description of the preferred embodiments of the present invention.

Detailed Description

For a further understanding of the invention, reference will now be made to the preferred embodiments of the invention by way of example, and it is to be understood that the description is intended to further illustrate features and advantages of the invention, and not to limit the scope of the claims.

The following materials or reagents, unless otherwise specified, are commercially available.