Human CYP2C9 and VKORC1 genetic polymorphism detection kit

A gene polymorphism and detection kit technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc. The effect of stable typing detection

Inactive Publication Date: 2015-12-23
WUHAN YZY MEDICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The purpose of the present invention is to overcome the deficiencies in the prior art and provide a detection kit for CYP2C9 and VKORC1 gene polymorphisms, so as to solve the problem of gene polymorphism detection when the sample size is small in the existing gene polymorphism detection technology Problems with unsatisfactory results

Method used

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  • Human CYP2C9 and VKORC1 genetic polymorphism detection kit
  • Human CYP2C9 and VKORC1 genetic polymorphism detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0071] Preparation of CYP2C9 and VKORC1 gene polymorphism detection kit of the present invention comprises the following steps:

[0072] 1. Primer and probe synthesis:

[0073] Design and synthesize 2 groups of specific primers SEQIDNO:1, SEQIDNO:2, SEQIDNO:3 and SEQIDNO:4; SEQIDNO:5 and SEQIDNO:6; 2 groups of specific probes SEQIDNO:7 and SEQIDNO:8, SEQIDNO:9 and SEQIDNO:10, and label FAM fluorescent group at the 5' end of SEQIDNO:7 and SEQIDNO:9, label NFQ-MGB non-luminescence quenching group at the 3' end, label at the 5' end of SEQIDNO:8 and SEQIDNO:10 VIC fluorescent group, 3' end labeled NFQ-MGB non-emission quenching group. The primers and probes were prepared into 100 μM stock solutions for storage.

[0074] 2. Prepare the internal standard system: design and synthesize a pair of internal standard primers for the human-derived gene GADPH, the primer pair sequences are SEQ ID NO: 11 and SEQ ID NO: 12; design and synthesize the internal standard probe, the probe is SE...

Embodiment 2

[0081] The CYP2C9 and VKORC1 gene polymorphism detection kit prepared in Example 1 was used to detect the samples to be tested.

[0082] In this embodiment, 40 cases of anticoagulated whole blood samples were collected, and genomic DNA was extracted from them, and the CYP2C9 and VKORC1 gene polymorphism detection kits obtained in Example 1 were used to detect the gene polymorphisms of CYP2C9 and VKORC1 in the samples to be tested .

[0083] 1. Genomic DNA extraction from blood samples

[0084] Take 300 μl of whole blood, add 900 μl of cell lysate CL, invert and mix well, let stand for 5 minutes, centrifuge at 10,000 rpm (11,500×g) for 1 minute, absorb the supernatant, leave the cell pellet, add to the cell pellet collected by centrifugation 200μl Buffer GS, shake until thoroughly mixed. Add 20 μl ProteinaseK solution and mix well. Add 200 μl buffer GB, mix thoroughly by inversion, place at 56°C for 10 minutes, invert and mix several times until the solution becomes clear (i...

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Abstract

The present invention provides a human CYP2C9 and VKORC1 genetic polymorphism detection kit. The kit includes a PCR buffer, dNTP, MgCl2, 2 groups of specific primers, 2 groups of specific probes, an internal standard system, HotStart Taq enzyme and UNG enzyme. The detection kit for detecting CYP2C9 and VKORC1 genetic polymorphism has the advantages of high specificity, high sensitivity, easy and fast operation, high-throughput, security, and objective interpretation of the results.

Description

technical field [0001] The invention relates to a kit for detecting polymorphisms of human CYP2C9 and VKORC1 genes, in particular to a kit for rapidly and effectively detecting polymorphisms of CYP2C9 and VKORC1 genes in a small amount of samples. Background technique [0002] Warfarin is currently the earliest, largest and most widely used oral anticoagulant in clinical practice, and has been used in anticoagulant therapy for many diseases, such as primary and secondary prevention of venous thromboembolic disease (VTE), atrial Prevention of tremor thromboembolism, valvular disease, artificial valve replacement, and intracardiac thrombosis, etc., but the clinical efficacy and adverse reactions vary greatly from individual to individual, and the dose is difficult to control, especially in the initial stage of warfarin anticoagulant therapy, which can easily lead to Serious bleeding complications. It is estimated that among patients taking warfarin, 15.2% of them experience b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6848C12Q1/6851C12Q2531/113C12Q2537/143C12Q2561/101C12Q2545/101C12Q2521/531C12Q2547/101
Inventor 周鹏飞蔡从利张喆王雯静薛伟沧李丽琼
Owner WUHAN YZY MEDICAL SCI & TECH
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