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Kit and method for drug-related gene typing detection of warfarin

A technology for genotyping and detection methods, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of low specificity and long detection time, and achieve high detection specificity, The effect of reducing renaturation and improving nano-aggregation effect

Inactive Publication Date: 2019-03-19
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, the technical problem to be solved by the present invention is that the genotyping detection kit related to warfarin medication in the prior art has a long detection time and low specificity, and then provides a fast, high specificity test kit for warfarin. Kits and detection methods for drug-related genotyping detection

Method used

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  • Kit and method for drug-related gene typing detection of warfarin
  • Kit and method for drug-related gene typing detection of warfarin
  • Kit and method for drug-related gene typing detection of warfarin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] This embodiment provides a nucleic acid amplification reagent, comprising the following concentrations of components:

[0081] High-fidelity DNA polymerase, 550mg / L;

[0082] dNTPs, 100g / L;

[0083] 10×PCR amplification buffer, 10 μL;

[0084] Nano silica, 110g / L;

[0085] Alkylphenol polyoxyethylene ether (APEO), 1mL / L;

[0086] Add double distilled water to 50 μL.

[0087] The above-mentioned nano silicon dioxide is spherical, 30nm.

[0088] The above-mentioned nano silicon dioxide is a commercially available product, such as purchased from Changtai Micro-Nano Chemical Factory in Shouguang City, Shandong Province.

Embodiment 2

[0090] This embodiment provides a nucleic acid amplification reagent, comprising the following concentrations of components:

[0091] Taq enzyme, 100mg / L;

[0092] dNTPs, 200g / L;

[0093] Tris-HCl, 0.5mL / L;

[0094] MgCl 2 , 40g / L;

[0095] KCl, 5g / L;

[0096] (NH 4 ) 2 SO 4 , 40g / L;

[0097] Nano silica, 20g / L;

[0098] Triton X-100, 5mL / L.

[0099] The above-mentioned nano silicon dioxide is spherical, 50nm.

[0100]Above-mentioned nano silicon dioxide, preparation method is as follows: ammoniacal liquor (concentration is 26wt%), water, dehydrated alcohol are placed in A bottle according to volume ratio 10: 0.5: 89.5, tetraethyl orthosilicate (TEOS) and no Water and ethanol are mixed according to the volume ratio of 15:85 and placed in bottle B. Mix the two bottles of solutions on a heating stirrer, stir and preheat to a reaction temperature of 100° C., and then mix them and add them to a three-neck flask with a thermal constant temperature heating magnetic stirr...

Embodiment 3

[0102] This embodiment provides a nucleic acid amplification reagent, comprising the following concentrations of components:

[0103] Taq enzyme, 500mg / L;

[0104] dNTPs, 50g / L;

[0105] Tris-HCl, 2mL / L;

[0106] MgCl 2 , 10g / L;

[0107] KCl, 20g / L;

[0108] (NH 4 ) 2 SO 4 , 5g / L;

[0109] Nano silica, 100g / L;

[0110] Triton X-100, 1 mL / L.

[0111] The above-mentioned nano silicon dioxide is spherical, 500nm.

[0112] Above-mentioned nano silicon dioxide, preparation method is as follows: ammoniacal liquor (concentration is 28wt%), water, dehydrated alcohol are placed in bottle A according to volume ratio 30: 2: 68, tetraethyl orthosilicate (TEOS) and no Water and ethanol are mixed according to the volume ratio of 30:70 and placed in bottle B. After mixing the two bottles of solutions, they were stirred on a heating stirrer and preheated to a reaction temperature of 120° C., and then mixed and added to a three-necked flask with a thermal constant temperature heati...

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Abstract

The invention belongs to the technical field of gene amplification detection, and particularly relates to a kit and a method for drug-related gene typing detection of warfarin. Use of nano-silica in gene amplification detection is initiatively found, and gene amplification reaction efficiency and reaction specificity can be remarkably improved, so that the nano-silica is applied to drug-related genotyping detection of the warfarin to solve the problems that drug-related genotyping for the warfarin is poor in specificity and low in detection speed, and a large number of samples cannot be rapidly detected in a high-specificity manner in the prior art.

Description

technical field [0001] The invention belongs to the technical field of gene amplification detection, and in particular relates to a detection kit and a detection method for genotyping related to warfarin medication. Background technique [0002] Warfarin (warfarin) is a coumarin oral anticoagulant commonly used in artificial valve replacement, thromboembolic disease and anticoagulant treatment of atrial fibrillation. Due to the narrow therapeutic window of warfarin, during clinical use, the stable treatment dose varies among different individuals. If the dose is insufficient, it will lead to the formation of thrombus, and if the dose is too large, it will increase the risk of bleeding. An estimated 15.2% of patients taking warfarin experience bleeding side effects each year, with fatal major bleeding accounting for 3.5%. The difference in the stable dose of warfarin among different individuals can reach more than 20 times. Pharmacogenetic studies have proved that the genet...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/6883C12Q2600/106C12Q2600/156C12Q2563/107
Inventor 周连群李金泽张芷齐李传宇姚佳张威郭振
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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