Kit for detecting warfarin sensitivity gene by using electrochemical gene sensor method

A kit and sensitivity technology, which is applied in the field of kits for detection of warfarin sensitivity genes by electrochemical gene sensor method, can solve the problems of limited influence of non-genetic factors and individual differences in warfarin dosage

Inactive Publication Date: 2014-03-12
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the degree of influence of non-genetic factors is relatively limited and is not the main reason for individual differences in warfarin dosage

Method used

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  • Kit for detecting warfarin sensitivity gene by using electrochemical gene sensor method
  • Kit for detecting warfarin sensitivity gene by using electrochemical gene sensor method
  • Kit for detecting warfarin sensitivity gene by using electrochemical gene sensor method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1: Development of Warfarin Sensitivity Gene Detection Reagent

[0079] 1. Design of primers and probes: Through sequence comparison and analysis of the warfarin-sensitive gene sequences on the human genome already in the Genebank database and related nucleic acid sequences reported in domestic and foreign published literature, the warfarin-sensitive The conserved fragment of the sex gene is the target site for amplification, and a highly conserved segment with no secondary structure is selected. According to the basic principles of primer and probe design, multiple pairs of primers and probes are manually designed using software.

[0080]2. Selection of samples: According to relevant domestic and foreign literature reports, samples such as human cells, anticoagulated whole blood, or tissues can be selected.

[0081] 3. Establishment and optimization of the reaction system

[0082] Preparation of samples: 6 cases of anticoagulated whole blood samples and 2 cases...

Embodiment 2

[0095] Embodiment 2: warfarin sensitivity gene detection kit (electrochemical gene sensor method) and its use

[0096] 1. Prepare a kit comprising the following components: Warfarin PCR reaction solution A (888 μL / tube) 1 tube, warfarin PCR reaction solution B (72 μL / tube) 1 tube, warfarin sensitive gene quality control (50μL / tube) 1 tube, blank control (50μL / tube) 1 tube, hybridization solution I (1680μL / tube) 1 tube, hybridization solution II (240μL / tube) 1 tube, hybridization solution III (480μL / tube) 1 tube , Electrochemical sensor (chip) (8 pcs / pack) 3 packs.

[0097] 2. Specimen collection, transportation and storage

[0098] 2.1 Applicable sample types: anticoagulated whole blood, etc.

[0099] 2.2 Sample collection and pretreatment (pay attention to aseptic operation): Draw 2 mL of venous blood from the subject with a disposable sterile syringe, inject it into a glass tube containing EDTA or sodium citrate anticoagulant, and immediately invert the glass tube gently f...

Embodiment 3

[0114] Example 3: Application of Warfarin Sensitivity Gene Detection Kit (Electrochemical Gene Sensor Method) to Detect Clinical Samples

[0115] All the experimental process was completed in the laboratory of the 307th Hospital of the Chinese People's Liberation Army, and the clinical samples were provided by the laboratory of the 307th Hospital of the Chinese People's Liberation Army, including anticoagulated whole blood, extracted DNA samples and other sample types, DNA extraction of specimens , PCR reaction and result analysis are carried out with reference to embodiment 2.

[0116] The detection results of the clinical samples are shown in the accompanying drawings: the genotypes of the warfarin drug sensitivity gene loci in the clinical samples can be visually determined from the detection reports of the two clinical samples (see the accompanying drawingsfigure 1 , figure 2 ).

[0117] figure 1 Explanation: The genotypes of the five warfarin-sensitive gene SNP sites i...

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Abstract

The invention provides a warfarin sensitivity gene detection kit (an electrochemical gene sensor method) which is a kit used for rapid detection of a warfarin sensitivity gene in a human genome by using electrochemical gene sensor technology. The kit is applicable to qualitative detection of *2, *3 and *5 allele types of the cytochrome CYP2C9 gene and 1639G>A and 1173C>T allele types of vitamin K epoxide reductase complex 1 (VKORC 1) in clinical specimens like human peripheral blood and a variety of tissue and cells; the above-mentioned genotypes are helpful for clinical discrimination of patients with increased drug susceptibility to warfarin and enable the dosage range of warfarin to be rapidly determined, curative effects to be guaranteed and the risk of hemorrhage to be reduced. Combination of warfarin gene detection results with INR monitoring enables the maintenance dose of warfarin to be effectively and rapidly adjusted, thereby lowering down the risk of hemorrhage while achieving curative effects.

Description

technical field [0001] The invention relates to a kit for detecting warfarin-sensitive genes by an electrochemical gene sensor method, in particular to a kit for rapidly detecting warfarin-sensitive genes in human genome by using an electrochemical gene sensor technology. This kit has high sensitivity and specificity. Through the kit of the present invention, rapid detection and analysis of farin-sensitive genes in the human genome in samples such as human cells, anticoagulated whole blood, or tissues can be realized. Background technique [0002] At the end of 1998, the American Association for the Advancement of Science listed electrochemical gene sensor technology as one of the top ten advances in natural science in 1998, which shows its significance in the history of science. It has shown great power with its ability to analyze thousands of genome information simultaneously, quickly and accurately. These applications mainly include gene expression detection, mutation d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6834C12Q1/6827C12Q2531/113C12Q2537/143C12Q2565/607
Inventor 陈华云李明肖湘文赵丽黄桃生
Owner DAAN GENE CO LTD
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