Kit for detecting curative effect of aspirin and detection method thereof

A technology of aspirin and a kit, which is applied to the kit for detecting the curative effect of aspirin and its detection field, which can solve problems such as interference with aspirin curative effect detection, inability to accurately detect samples, and false negative detection kits, so as to improve sensitivity and accuracy, reduce The generation of false negatives, the effect of the simple detection method

Active Publication Date: 2015-08-19
华阴市锦前程药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem that the 11-dh-TXB2 detection kit in the prior art is prone to produce false negatives, the inventor has carried out a large number of test analyzes on the detection kit and detection samples, and unexpectedly found that: the preservation of urine samples , usually need to add preservatives (such as 0.01% NaN3), and NaN 3 It will inhibit the activity of horseradish peroxidase (HRP) and interfere with the detection of aspirin efficacy. The existing detection kits cannot accurately detect samples containing NaN3, resulting in false negative detection and analysis results

Method used

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  • Kit for detecting curative effect of aspirin and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of 11-dehydrothromboxane B2 monoclonal antibody

[0022] The preparation of 11-dehydrothromboxane B2 monoclonal antibody comprises the following steps: A) Using 7-week-old Balb / c mice as immunized animals, the purified 11-dehydrothromboxane B2 cross-linked protein is used as immunogen, and immunized The dose is 60 μg / rat. For the first immunization, mix and emulsify the immunogen with the same amount of Freund's complete adjuvant, and inject it subcutaneously at multiple points on the back of the neck. After an interval of 2 weeks, take the same dose of immunogen plus the same amount of Freund's incomplete adjuvant. Splenocytes were mixed with the Sp2 / 0 myeloma cells at a ratio of 5:1, and the hybridoma cells were screened using HAT medium, and then the positive cells were expanded Cultivate, produce and secrete monoclonal antibodies, collect culture supernatant, centrifuge to remove cells and their debris, and obtain.

[0023] The prepared 11-de...

Embodiment 2

[0024] Example 2 Preparation of ELISA plate coated with 11-dehydrothromboxane B2 antibody

[0025] 04μg / The antibody diluent of ml concentration; B) add 100 μ L antibody diluent to the microtiter plate, after covering for 3 hours, wash with deionized water containing 4% Tween-20, pat dry; C) add to the microtiter plate Blocking solution containing 10% skimmed milk powder, after blocking for 2 hours, pour off the liquid in the well, and vacuum dry.

Embodiment 3

[0026] Example 3 Kit for detecting the curative effect of aspirin

[0027] A kit for detecting the efficacy of aspirin, including 11-dehydrothromboxane B2 antibody-coated microtiter plate, horseradish peroxidase-labeled 11-dehydrothromboxane B2 antibody, and serial concentrations of 11-dehydrothromboxane B2 Standard solution, substrate chromogenic solution and 0.3% trypsin dilution.

[0028] The kit for detecting the curative effect of aspirin provided by the present invention is stored in cold storage at 4°C, and can be guaranteed for more than one year. Before use, it is taken out from the cold storage environment and equilibrated at room temperature for more than 15 minutes. All reagents must be shaken before use.

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Abstract

The invention provides a kit for detecting the curative effect of aspirin, belonging to the technical field of biology. The kit comprises a 11-dehydrogenated thromboxane B2 antibody-coated elisa plate, a horseradish peroxidase-marked 11-dehydrogenated thromboxane B2 antibody, 11-dehydrogenated thromboxane B2 standard solution, substrate color developing liquid and an interference shielding agent. The kit for detecting the curative effect of the aspirin comprises the interference shielding agent, so the interference of NaN3 in a detection sample can be effectively avoided, the generation of false negative is reduced, and the detection sensitivity and accuracy for the curative effect of the aspirin are improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a kit for detecting the curative effect of aspirin and a detection method thereof. Background technique [0002] Aspirin can inhibit the generation of thromboxane A2 (TXA2), and is widely used in the prevention and treatment of cardiovascular diseases. Aspirin resistance usually refers to the failure of aspirin treatment to achieve the expected biological effects (such as inhibition of platelet aggregation, inhibition of thromboxane biosynthesis, and prolongation of bleeding time) or failure to prevent atherosclerotic thrombosis events. Timely detection of aspirin resistance is crucial to improving the efficacy of patients with cardiovascular and cerebrovascular diseases. [0003] The paper titled "Aspirin Resistance and Its Experimental Monitoring" published by Zhang Liwei and others in "Diagnostics Theory and Practice" introduced aspirin resistance and its corresponding detection ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543
CPCG01N33/5008G01N33/543
Inventor 陈立国孙丽华
Owner 华阴市锦前程药业有限公司
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