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Primer, probe and detection kit for detecting polymorphism of CYP2C19 genes

A CYP2C19, polymorphism detection technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of low accuracy, poor repeatability, false negative, etc. Intuitive effect of detection results

Inactive Publication Date: 2017-10-03
庄江兴 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the advantages of high throughput and other advantages, gene chip technology has been widely used in SNP detection, but there are also disadvantages such as difficult control of detection conditions, poor repeatability, low accuracy, prone to false positive and false negative results, and low sensitivity.

Method used

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  • Primer, probe and detection kit for detecting polymorphism of CYP2C19 genes
  • Primer, probe and detection kit for detecting polymorphism of CYP2C19 genes
  • Primer, probe and detection kit for detecting polymorphism of CYP2C19 genes

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Embodiment Construction

[0038] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.

[0039] 1. Materials

[0040] 1. Instruments: real-time fluorescent PCR instrument, pipette, centrifuge.

[0041] 2. Design of primers and probes: The present invention simultaneously designs two pairs of high-efficiency specific primers for the detection of target genes, and simultaneously designs two pairs of probes for detection of corresponding CYP2C19 gene rs4244285 and rs4986893 genetic polymorphic sites. The primers and probe series are as follows:

[0042] Primers:

[0043] CYP2C19*2(rs4244285)-F:5'-CCAGAGCTTGGCATATTGTATCT-3'

[0044] CYP2C19*2(rs4244285)-R:5'-TGTCCATCGATTCTTGGTGTTCT-3'

[0045] CYP2C19*3(rs4986893)-F:5'-TCTGCTCCATTATTTTCCAGAAACG-3'

[0046] CYP2C19*3(rs4986893)-F:5'-CTTCAGGGCTTGGTCAATATAGAAT-3'.

[0047] Probe:

[0048] FAM-5'-GATGAAATCGGCTCCCGC-3'BHQ1

[0049] HEX-5'-GATGAAATCGACTCCCGCAG-3'-BHQ1

[0050] ROX-5'-AGACA...

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Abstract

The invention discloses a primer, a probe and a detection kit for detecting polymorphism of CYP2C19 genes, and comprises an amplification primer pair and a fluorescent probe for detecting the polymorphism of rs4244285 and rs4986893 of the CYP2C19 genes. By utilizing the probe marked with different fluorophores, single-tube enclosure is realized and four co-existence gene types of rs4244285 and rs4986893 of the CYP2C19 genes in samples are detected at the same time, and the defect of low detection flux of a conventional fluorescence method is overcome and cross contamination is reduced; the operation is simple and convenient, and the detection is quick; the detection result is intuitive, all gene types in samples to be detected can be qualitatively judged according to amplification curves produced by different fluorescent signals, and a cumbersome analysis process is not needed. The kit has the advantages of quickness, correctness, high flux, low cost, no pollution and the like, and can be used as a medication guide of medicines such as clopidogrel, S-mephenytoin, omeprazole, voriconazole, diazepam and nordiazepam.

Description

technical field [0001] The present invention relates to nucleic acid detection in vitro, in particular to the detection of CYP2C19 genes CYP2C19*2 (rs4244285, c.681G>A) and CYP2C19*3 (rs4986893, c.636G>A) using fluorescence quantitative PCR (FQ-PCR) technology Primers, probes and detection kits for polymorphisms. Background technique [0002] CYP2C19 is a very important drug-metabolizing enzyme. The human CYP2C19 gene is located on chromosome 10, with a length of about 90.21kb, including 9 exons and 8 introns. CYP2C19 genetic polymorphisms can lead to individual differences in enzyme activity, resulting in individual differences in blood drug concentrations. About 2% of clinical drugs are catalyzed by CYP2C19 metabolism, including the metabolism of clopidogrel, S-mephenytoin, omeprazole, voriconazole, diazepam, and norazepam. Individual differences caused by CYP2C19 genetic variation lead to ultrarapid metabolizers (UM), extensive metabolizers (EM), intermediate meta...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/106C12Q2600/156C12Q2600/172C12Q2531/113C12Q2563/107
Inventor 庄江兴
Owner 庄江兴
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