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Human CYP2C19 gene polymorphism detection primers, probes and kit, and application of human CYP2C19 gene polymorphism detection primers

A technology of CYP2C19 and gene polymorphism, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems that are not conducive to reducing product development costs, limiting detection sensitivity, and reducing primer concentration. , to achieve the effects of reducing the difference in amplification efficiency, avoiding sample cross-contamination, and high sensitivity

Inactive Publication Date: 2020-07-10
安徽同科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Reducing the primer concentration can reduce the primer dimer at the same time, but it limits the detection sensitivity; the experiment of adjusting the ratio of upstream and downstream primers is cumbersome, which is not conducive to reducing product development costs

Method used

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  • Human CYP2C19 gene polymorphism detection primers, probes and kit, and application of human CYP2C19 gene polymorphism detection primers
  • Human CYP2C19 gene polymorphism detection primers, probes and kit, and application of human CYP2C19 gene polymorphism detection primers
  • Human CYP2C19 gene polymorphism detection primers, probes and kit, and application of human CYP2C19 gene polymorphism detection primers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1: Human CYP2C19 Gene Polymorphism Detection Kit

[0063] The test kit described in this embodiment includes the following components:

[0064] Public primers and probes: public primer UP1 (nucleotide sequence shown in SEQ ID NO.1), β-globin specific upstream primer globin F (nucleotide sequence shown in SEQ ID NO.14), β-globin specific Sex downstream primer globin R (nucleotide sequence shown in SEQ ID NO.15), β-globin specific probe globin probe (nucleotide sequence shown in SEQ ID NO.16);

[0065] Specific Primers and Probes:

[0066] CYP2C19 681G-specific upstream primer C2W1F (nucleotide sequence shown in SEQ ID NO.2), CYP2C19 681A-specific downstream primer C2M1F (nucleotide sequence shown in SEQ ID NO.3) for the qualitative detection of CYP2C19 681G>A gene polymorphism nucleotide sequence), CYP2C19 681 downstream primer C2R1 (nucleotide sequence shown in SEQ ID NO.4), CYP2C19 681 specific probe C2probe (nucleotide sequence shown in SEQ ID NO.5);

[006...

Embodiment 2

[0087] Embodiment 2: Human CYP2C19 gene polymorphism detection method

[0088] Use the test kit of embodiment 1 to detect, comprise the following steps:

[0089] 1. Preparation of DNA

[0090] PCR amplification obtains products containing the gene sequences of SEQ ID NO.17, SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20, SEQ ID NO.21, and SEQ ID NO.22, and is connected to pMD18 after electrophoresis purification -T vector, transformed into Escherichia coli and then shaken to extract the plasmid and send it for sequencing verification, see the sequencing results figure 1 - Figure 6, where the SNP site is marked with a black border, and the Nano drop2000 detection concentration is diluted to 10 with 1 times TE buffer (pH=8.0) 6 Copies / μL are reserved and named as V1, V2, V3, V4, V5, V6 respectively.

[0091] 1.1 Preparation of plasmid reference products, the names and components of the reference products are as follows:

[0092]

[0093]

[0094] 1.2 Storage of reference pro...

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Abstract

The present invention provides human CYP2C19 gene polymorphism detection primers, human CYP2C19 gene polymorphism detection probes and a human CYP2C19 gene polymorphism detection kit and an application of the human CYP2C19 gene polymorphism detection primers. The human CYP2C19 gene polymorphism detection primers comprise specific primers for G681A detection, specific primers for G636A detection, specific primers for transcription upstream C-806T detection, specific primers for internal reference gene beta-globin detection and a common primer for PCR reactions. The human CYP2C19 gene polymorphism detection primers increase compatibility of multiple PCR reactions, also ensure consistency of amplification efficiency and reduce result deviations caused by difference of amplification efficiency.

Description

technical field [0001] The invention relates to a human CYP2C19 gene polymorphism primer, a probe kit and applications thereof, which belong to the field of biotechnology. Background technique [0002] Clopidogrel is an antiplatelet drug widely used in cardiovascular diseases. Platelet activation and aggregation are the key factors for arterial thrombosis after acute coronary syndrome and percutaneous coronary intervention (PCI). Antiplatelet therapy combining the two drugs clopidogrel and aspirin is essential for the prevention of ischemic events in patients with acute coronary syndrome and after PCI. However, after receiving this antiplatelet therapy, about 10% of patients still experience cardiovascular events such as stent thrombosis, death, and stroke. The reason is that there are differences in the response of different patients to clopidogrel, and among many influencing factors, gene polymorphism plays a decisive role. [0003] Clopidogrel forms a thiol derivative ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2531/113C12Q2537/143C12Q2563/107C12Q2545/101
Inventor 徐阳宋冬梅何胜祥陈林峰
Owner 安徽同科生物科技有限公司
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