CYP2C19 gene polymorphyism detection kit and detection method thereof

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...

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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 , false negative results; 4) low sensitivity: the chip method requires a large amount of nucleic acid, and generally requires multiple PCR amplification first. As a result, the efficiency of amplifying the target fragments is different, which in turn affects the sensitivity of detection; 5) Due to the variety of chips, it is difficult to formulate a unified quality control standard

[0011] At present, there are no relevant research reports on the CYP2C19 gene polymorphism detection kit and its detection method based on the GeXP multiple gene expression genetic analysis system at home and abroad

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