Human CYP2C19 gene polymorphism detection specific primer and kit

A CYP2C19, gene polymorphism technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of unsatisfactory gene polymorphism detection effect, etc., to achieve true and credible results , Reduce pollution and stabilize the effect of typing detection

Inactive Publication Date: 2015-10-21
WUHAN YZY MEDICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the embodiments of the present invention is to overcome the deficiencies in the prior art, and provide a detection primer and kit for human CYP2C19 gene polymorphism, so

Method used

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  • Human CYP2C19 gene polymorphism detection specific primer and kit
  • Human CYP2C19 gene polymorphism detection specific primer and kit
  • Human CYP2C19 gene polymorphism detection specific primer and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Preparation of human CYP2C19 gene polymorphism detection kit of the present invention comprises the following steps:

[0056] 1. Primer and probe synthesis:

[0057] Design and synthesize three sets of specific primers: SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9; 3 sets of specific probes SEQ ID NO:10 and SEQ ID NO:11, SEQ ID NO:12 and SEQ ID NO:13, SEQ ID NO:14 and SEQ ID NO:15, and label the FAM fluorescent group at the 5' end of SEQ ID NO:10, SEQ ID NO:12 and SEQ ID NO:14, and label the NFQ-MGB non-luminescent quenching group at the 3' end, in SEQ ID NO:14 The 5' end of NO:11, SEQ ID NO:13 and SEQ ID NO:15 is labeled with VIC fluorescent group, and the 3' end is labeled with NFQ-MGB non-emission quenching group. The primers and probes were prepared into 100 μM stock solutions for storage.

[0058] 2. Prepare the internal standard system: design and synthesize a pair of internal stand...

Embodiment 2

[0071] The human CYP2C19 gene polymorphism detection kit prepared in Example 1 was used to detect the samples to be tested.

[0072] In this embodiment, the anticoagulated whole blood samples of 40 cardiovascular patients were collected, and genomic DNA was extracted therefrom, and the CYP2C19 gene polymorphism of the sample to be tested was detected with the human CYP2C19 gene polymorphism detection kit obtained in Example 1 Condition.

[0073] 1. Genomic DNA extraction from blood samples

[0074]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 Proteinase K solution and mix well. Add 200 μl buffer GB, mix well by inversion, place at 56°C for 10 minutes, invert and mix several times during this time, the solution shoul...

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Abstract

The invention belongs to the field of biotechnologies and medicine, and provides a specific primer used for human CYP2C19 gene polymorphism detection. The nucleotide sequences of the primer are shown in SEQ ID NO:1-9. The invention further provides a kit for human CYP2C19 gene polymorphism detection. The primer and the kit are used for human CYP2C19 gene polymorphism detection and have the advantages of being high in specificity and sensitivity, easy and fast to operate and safe and having high flux, and result interpretation is objective.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a human CYP2C19 gene polymorphism detection specific primer and kit. Background technique [0002] CYP2C19 is one of the most important drug-metabolizing enzymes in the CYP450 family. Many endogenous substrates and about 2% of clinical drugs are metabolized by it. Studies have found that CYP2C19 can affect the metabolism of many important clinically used drugs such as clopidogrel, omeprazole, diazepam, and phenytoin, and its genetic polymorphism is the cause of different metabolism of the same drug between individuals and races. One of the reasons for the ability. In 2010, the US FDA required that the relationship between CYP2C19 and curative effect be indicated on the drug label of clopidogrel, and it was recommended to detect the CYP2C19 genotype before use. [0003] CYP2C19 gene wild type is CYP2C19*1 / *1 type, CYP2C19*2 type, CYP2C19*3 type and CYP2C19*17 are the more common all...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
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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