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Preparation method of platelet quality control product and application of prepared platelet quality control product

A technology for platelet-rich plasma and platelets, which is applied in the field of in vitro diagnosis, can solve the problems that the detection of platelet adhesion function and aggregation function cannot be applied, and achieves the effects of light weight, simplified preparation steps and reduced production cost.

Pending Publication Date: 2020-08-18
上海原科实业发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the simulated platelets are only used for counting and do not have adhesion and aggregation functions, platelet quality control products can only be used for platelet counting items, but not for platelet adhesion function and aggregation function testing items.

Method used

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  • Preparation method of platelet quality control product and application of prepared platelet quality control product
  • Preparation method of platelet quality control product and application of prepared platelet quality control product
  • Preparation method of platelet quality control product and application of prepared platelet quality control product

Examples

Experimental program
Comparison scheme
Effect test

preparation Embodiment 1

[0033] S1, collect 1000mL fresh bovine whole blood, add 20000iU heparin anticoagulant, and centrifuge at 1000rpm for 5 minutes. After centrifugation, the upper layer is platelet-rich plasma, and the lower layer is packed white blood cells and red blood cells;

[0034] S2, transfer 100mL platelet-rich plasma into two 50ml centrifuge tubes, 40mL in each tube;

[0035] S3, centrifuge the plasma in two 50mL centrifuge tubes at 1500rpm for 10 minutes, and collect the platelet-poor plasma in the upper layer;

[0036] S4, weigh 0.450g NaCl, pipette 0.5mL DMSO and add it into purified water to prepare 50mL washing solution;

[0037] S5, weigh 1.197g HEPES, add 0.200g NaOH into purified water to configure 500mL of 10mmol / L HEPES buffer solution, and measure the pH value to be 7.3;

[0038] S6, measure 100mL of the 10mmol / L HEPES buffer prepared in S5, weigh 1.892g of fucose, 0.584g of NaCl, 0.373g of KCl, and 0.7mg of tubulin inhibitor into the HEPES buffer to prepare a loading soluti...

preparation Embodiment 2

[0048] S1, collect 1000mL fresh bovine whole blood, add 3.8g sodium citrate anticoagulant, and centrifuge at 1000rpm for 5 minutes. After centrifugation, the upper layer is platelet-rich plasma, and the lower layer is packed white blood cells and red blood cells;

[0049] S2, transfer 100mL platelet-rich plasma into two 50ml centrifuge tubes, 40mL in each tube;

[0050] S3, centrifuge the plasma in two 50mL centrifuge tubes at 1000rpm for 5 minutes, and collect the platelet-poor plasma in the upper layer;

[0051] S4, weigh 0.450g NaCl, pipette 0.5mL DMSO into purified water, and prepare 50mL washing solution;

[0052]S5, weigh 0.599g HEPES, add 0.100g NaOH into purified water to prepare 500mL 5mmol / L HEPES buffer solution, and measure the pH value to be 7.2;

[0053] S6, measure 100 mL of 5 mmol / L HEPES buffer, weigh 1.135 g of fucose, 0.701 g of NaCl, 0.298 g of KCl, and 0.5 mg of tubulin inhibitor into the HEPES buffer, and prepare 100 mL of loading solution;

[0054] S7,...

preparation Embodiment 3

[0063] S1, collect 1000mL fresh bovine whole blood, add 3.8g sodium citrate anticoagulant, and centrifuge at 1000rpm for 5 minutes. After centrifugation, the upper layer is platelet-rich plasma, and the lower layer is packed white blood cells and red blood cells;

[0064] S2, transfer 100mL platelet-rich plasma into two 50ml centrifuge tubes, 40mL in each tube;

[0065] S3, centrifuge the plasma in two 50mL centrifuge tubes at 1500rpm for 5 minutes, and collect the platelet-poor plasma in the upper layer;

[0066] S4, weigh 0.450g NaCl, pipette 0.5mL DMSO into purified water, and prepare 50mL washing solution;

[0067] S5, weigh 1.796g HEPES, add 0.30NaOH into purified water to prepare 500mL of 15mmol / L HEPES buffer, and measure the pH value to be 7.4;

[0068] S6, measure 100 mL of 15 mmol / L HEPES buffer, weigh 2.648 g of fucose, 0.468 g of NaCl, 0.447 g of KCl, and 0.9 mg of tubulin inhibitor into the HEPES buffer, and prepare 100 mL of loading solution;

[0069] S7, add 1...

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Abstract

The invention discloses a preparation method of a novel platelet quality control product and application of the platelet quality control product prepared by the method. The method comprises the following steps: (1) adding an anticoagulant into animal whole blood, and performing centrifuging to obtain platelet-rich plasma; (2) centrifuging the platelet-rich plasma again to obtain upper-layer platelet-depleted plasma and bottom platelets, washing and centrifuging the bottom platelets for multiple times to obtain concentrated platelets, adding a loading solution, performing water bath at 37 DEG C, and performing centrifuging to obtain platelets loaded with a protective agent; and (3) adding a freeze-drying buffer solution into the platelets loaded with the protective agent, performing pre-freezing for 18-36 hours at the temperature of-60 DEG C to-80 DEG C, and then carrying out freeze-drying; and storing the freeze-dried powder at 2-8 DEG C. The platelet quality control product prepared by the method not only can be used for platelet counting, but also can be suitable for platelet in-vitro adhesion function and aggregation function detection.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis, and in particular relates to a preparation method of a platelet quality control product and an application of the prepared platelet quality control product. Background technique [0002] Cardiovascular and cerebrovascular diseases have become the number one killer that endangers human health. Its pathological basis is arteriosclerosis and its secondary thrombosis and embolism. Platelets play an important role in the process of thrombus formation. Antiplatelet therapy is an important measure for the primary and secondary prevention of such diseases, and platelet function testing is an indispensable means in the prognosis and treatment of the disease. Therefore, it is particularly important to carry out quality control in the laboratory for platelet function testing. Platelet detection mainly includes platelet count, platelet adhesion function test and platelet aggregation function test. However...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 王连升李红梅
Owner 上海原科实业发展有限公司
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