CYP2C19 gene polymorphyism detection kit and detection method thereof

A technology for CYP2C19 gene polymorphism, applied in biochemical equipment and methods, measuring devices, microbial determination/inspection, etc. Aiming at complex problems such as synthesis and fixation, it achieves the effect of saving detection cost, fast cost, and solving easy pollution

Active Publication Date: 2013-05-01
NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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  • Abstract
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Problems solved by technology

However, there are also the following disadvantages: 1) The hybridization kinetics of different SNP sites are different, and it is difficult to control the conditions for simultaneous detection of multiple sites; 2) The technology is expensive and complicated: each sample requires a chip, and the cost is greater than ￥1000/sample, which is not conducive to large-scale promotion; the synthesis and fixation of probes are more complicated, especially the production of high-density probe arrays, which is the main speed-limiting step; 3) Poor repeatability, low accuracy, and prone to false positives

Method used

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  • CYP2C19 gene polymorphyism detection kit and detection method thereof
  • CYP2C19 gene polymorphyism detection kit and detection method thereof
  • CYP2C19 gene polymorphyism detection kit and detection method thereof

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specific Embodiment 2

[0049] A kind of CYP2C19 gene polymorphism detection method of the present invention, concrete steps are as follows:

[0050] 1. Production of a CYP2C19 gene polymorphism detection kit based on the GeXP multiple gene expression genetic analysis system, the components included in the kit are the same as in the above-mentioned embodiment 1;

[0051] 2. Collection and extraction of DNA samples

[0052] Put the buccal swab scraped from the oral epithelial cells into 300 μL DNA lysis buffer, and treat it in a constant temperature mixer at 95°C and 1000 rpm for 5 minutes, then take it out, leave it at room temperature until it cools down, and then put it in the sample Add 30 μL extraction buffer, mix well, and centrifuge at 12,000g for 5 minutes. The obtained supernatant is the DNA sample template for PCR;

[0053] 3. Use the extracted nucleic acid as a template for PCR reaction

[0054] 1) Add reagents and samples (PCR plate, see Table 3) to the 96-well sample plate / eight tubes i...

specific Embodiment 3

[0069] Detection Kit Sensitivity and Specificity Analysis

[0070] Sensitivity analysis: After diluting the positive control substance according to a certain copy number ratio, it is detected by PCR amplification and capillary electrophoresis until no signal is detected. The copy number is the lowest detection line, which is the sensitivity of the kit. The kit has a sensitivity of 50 copies.

[0071] Specificity analysis: Single-plex PCR amplification is detected as a single peak of the target fragment size by capillary electrophoresis.

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Abstract

The invention discloses a CYP2C19 gene polymorphyism detection kit and a detection method thereof. The kit comprises ultrapure water, a solution X, a 10*PCR (polymerase chain reaction) buffer solution, PCR primers, a 25mM magnesium chloride solution, DNA (deoxyribonucleic acid) polymerase and a positive control, and is characterized in that the PCR primers include a different genotypic forward and reverse amplification primer, an internal DNA reference forward and reverse amplification primer and an internal reaction reference forward and reverse amplification primer on three SNP (single nucleotide polymorphism) sites of a CYP2C19 gene; and gene sequences of the primers are shown as SEQ ID NO. 1-NO. 13 (sequence identifier number 1-number 13). The detection method comprises the steps of collecting a sample, extracting nucleic acid, taking nucleic acid of a patient as a template to conduct PCR reaction, and using a GeXP genetic analyzer to conduct capillary electrophoretic separation on the sample. The detection kit and the detection method have the advantages of high specificity, high sensitivity, high flux, high reliability, low cost and no false negative results.

Description

technical field [0001] The invention relates to a multiple gene detection kit and a detection method thereof, in particular to a CYP2C19 gene polymorphism detection kit and a detection method thereof. Background technique [0002] Cytochrome P450 (CYP) enzymes play an extremely important role in drug metabolism. CYP2C19 (S-Mephenytoin Hydroxylase) is one of the dominant enzymes in the CYP enzyme system. Its genetic polymorphism affects the metabolism of many clinical drugs, and its activity has obvious individual differences. Drug therapeutic effect and adverse reactions and drug toxicity have an important impact. The FDA has listed 17 drugs as drugs that doctors need to refer to the patient's CYP2C19 gene polymorphism information. Clinically, the detection of CYP2C19 gene polymorphism can provide the basis for individualized medicine. [0003] At present, there are several CYP2C19 genotype detection kits at home and abroad, and the leading method is the DNA microarray ch...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N27/447
Inventor 南丽孙婷婷吴勇
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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