Aspirin individualized drug gene detection kit and detection method

An aspirin and gene detection technology, which is applied in the field of detection kits for the detection of aspirin personalized medicine gene detection kits, and achieves high accuracy, accurate treatment and prevention of cardiovascular events, and the effects of precise treatment and prevention of cardiovascular events.

Pending Publication Date: 2019-01-15
中科基因生物科技(江苏)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The existing technology mainly has the following technical problems: lack of a gene detection kit for aspirin

Method used

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  • Aspirin individualized drug gene detection kit and detection method
  • Aspirin individualized drug gene detection kit and detection method
  • Aspirin individualized drug gene detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Genetic detection kit for one-person detection of aspirin individualized medicine, including the following components:

[0046] (1) 4 pairs of primers for amplification and sequencing of PTGS1 (-842A>G), ITGA2 (759C>T), ITGB3 (176T>C) and PEAR1 (-9-3996G>A), each primer 1OD.

[0047] The nucleotide sequences of each primer are shown in SEQ NO.5-12, see Table 1 for details:

[0048] Table 1: Primer sequences

[0049]

[0050] (2) PCR amplification reagent: 15 μL of Taq enzyme mixture (dNTP, 10×PCR reaction buffer, MgCl 2 ) (purchased from Novozyme);

[0051] (3) PCR product purification reagent: 3.2 μL SAP enzyme mixture (SAP enzyme, ExoI enzyme, deionized water) (purchased from Novizyme);

[0052] (4) Sequencing reagents: 4.4 μL BigDye mix (Bigdye, 5×seq), 5 μL EDTA, 60 μL ethanol solution (100%), 200 μL ethanol solution (70%), 16 μL Hi-Di.

[0053] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

Embodiment 2

[0055] Using the one-person detection aspirin individualized drug gene detection kit in Example 1, the steps are as follows:

[0056] (1) extract the genomic DNA of the sample;

[0057] (2) PCR amplification of PTGS1(-842A>G), ITGA2(759C>T), ITGB3(176T>C) and PEAR1(-9-3996G>A) loci

[0058] For PTGS1 (-842A>G), ITGA2 (759C>T), ITGB3 (176T>C) and PEAR1 (-9-3996G>A) sites, design primers according to Table 1 of Example 1 for PCR amplification.

[0059] Each reaction system has a total volume of 16 μL, including 7.5 μL of Taq enzyme mixture (dNTP, 10×PCR reaction buffer, MgCl 2 ), deionized water 5.5 μL, primer pair (10uM) 1 μL, genomic DNA (100ng / ul) 1ul. The reaction conditions are 95°C for 15 minutes, 14 cycles (-0.5°C / cycle) of 94°C for 30 seconds, 63°C for 30 seconds, and 72°C for 45 seconds; then 30 cycles of 94°C for 30 seconds and 56°C for 30 seconds , 72°C for 45 seconds, and finally 72°C for 10 minutes.

[0060] (3) Purification of PCR products

[0061] Each reacti...

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Abstract

A Gene detection kit for detect individualized aspirin use is characterize in that that kit comprises the following components: PTGS1 (-842A) G), ITGA2 (759C) T), ITGB3 (176T) C) and PEAR1 (-9-3996G)site amplification and sequence primer, PCR amplification reagent, PCR product purification reagent and DNA sequencing reagent, that nucleotide sequence of each prim is as follows: SEQ NO. 5-12 as shown in table 1. The invention has the following technical effects: a gene detection kit for individualized aspirin medication of Chinese population is provided, and the sensitivity is high and the accuracy is high. The kit is used for PTGS1 (-842A) G), ITGA2 (759C) T), ITGB3 (176T) C) and PEAR1 (-9-3996G) A) genotyping, which can detect genetic differences between individuals and determine the riskof aspirin resistance in advance, so that individualized antiplatelet therapy can be used to achieve accurate treatment and prevent cardiovascular events.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a gene detection kit and a detection method for detecting aspirin individualized drug use. Background technique [0002] Aspirin is an antipyretic and analgesic non-steroidal anti-inflammatory drug, which mainly produces antipyretic, analgesic and anti-inflammatory effects by inhibiting the synthesis of prostaglandins, bradykinin, and histamine. The mechanism of antipyretic action is to act on the hypothalamic thermoregulatory center to dilate peripheral blood vessels, increase skin blood flow, and increase heat dissipation to achieve a cooling effect. Its analgesic effect is a peripheral analgesic; aspirin is also a commonly used anti-platelet aggregation drug. It inhibits platelet cyclooxygenase, reduces the production of prostaglandins, and inhibits platelet aggregation. It is widely used clinically to reduce the risk of acute myocardial infarction in suspected...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2535/122
Inventor 姜昕王斐陆军景丹丹
Owner 中科基因生物科技(江苏)有限公司
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