Primers, probes, kit and method for detecting polymorphism of human CYP2C19 gene

Abstract

The invention belongs to the technical field of gene engineering and discloses a composition of primers and probes for detecting polymorphism of human CYP2C19 gene, a kit containing the composition of the primers and the probes, and a fluorescence PCR method for detecting the polymorphism of the human CYP2C19 gene by using the composition of the primers and the probes or the kit. Based on a TaqMan fluorescence PCR technology, the primers, the probes and the kit are simple and rapid, and are high in sensitivity; in addition, through reasonable collocation of the primers and the probes, the interaction between the primers, the interaction between the primers and the probes and the interaction between the probes can be effectively avoided; the detection errors are reduced. When the primers, the probes and the kit, provided by the invention, are used for detecting the polymorphism of the human CYP2C19 gene, the primers, the probes and the kit has the advantages that the sensitivity is high, the specificity is high, the operation is simple, rapid and safe, the result is simply and intuitively determined and read, and blood samples or dried blood spots samples on filter paper can be directly used as templates.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to primers, probes, kits and methods for detecting human CYP2C19 gene polymorphisms. Background technique [0002] CYP2C19 is an important member of the second subfamily of CYP450 enzymes and an important drug-metabolizing enzyme in the human body. About 2% of clinical drugs are metabolized by it. At present, the US FDA has announced that there are 19 drugs that use CYP2C19 as a pharmacogenomic biomarker, including clopidogrel, S-mephenytoin, omeprazole, voriconazole, diazepam, and norazepam. Gene polymorphisms of CYP2C19 can lead to individual differences in enzyme activity, causing the population to appear as an extensive metabolizer (EM), an intermediate metabolizer (IM) and a poor metabolizer (PM). There are many polymorphic sites in the CYP2C19 gene. The allelic types carried by Orientals are CYP2C19*2, CYP2C19*3 and CYP2C19*17. It can cause the loss of the enzyme...

AI Technical Summary

Problems solved by technology

Although the sequencing method is the "gold standard" for gene polymorphism detection, it requires a long detection time, complicated operation, and low sensitivity; the PCR-gene chip method has high operational requirements, and hybridization and washing must be protected from light operation, and the chip is expensive; the PCR-melting curve method is non-specific and cannot precisely locate the position of the gene change, and the difference in Tm caused by a single base difference is not very obvious, which usually results in inconspicuous peaks on the melting curve Therefore, there are certain difficulties in the clinical promotion of these three methods

The traditional fluorescent quantitative PCR method is widely used clinically. This method has certain requirements on the number of samples and the distribution of polymorphisms. When the sample size is small, it cannot accurately detect the polymorphisms of the samples. It cannot meet the requirements of high efficiency, accuracy, simplicity, etc. At the same time, it also needs to perform cumbersome and complicated operation procedures such as genomic DNA extraction on the sample, so there is an urgent need for a fast, effective, accurate detection method that does not require DNA extraction

Images