Composition for detecting CYP2C19 gene polymorphism and its application

Abstract

The invention discloses a composition for detecting CYP2C19 gene polymorphism and its application. The single nucleotide polymorphism site in the CYP2C19 gene is detected through FAM, HEX, ROX (modified) three-channel multiple PCR reaction. In order to improve the detection simplicity and specificity, one-tube parting detection is realized through the technical manner of a probe parting, thus the whole operation and the reaction process are simplified; on this basis, locked nucleic acid modification and second-grade structure modification are performed on the probe to increase the Tm value of the probe, and further improve the probe combining specificity; finally, the lowest detection limit, accuracy and the specificity of the whole kit are improved through adjusting the mixture ratio of the primer pair, dosage of Taq enzyme, dosage of magnesium ion and others; the lowest detection concentration of the kit can reach 1 mg / mu l; the accuracy and the specificity can reach 100%.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a technology for detecting CYP2C19 gene polymorphism. Background technique [0002] Clopidogrel, a thienopyridine antiplatelet drug, is a second-generation ADP receptor antagonist. Clopidogrel is an inactive prodrug that needs to be activated by the cytochrome P450 enzyme system in liver cells, about 85% of which are esterified into inactive metabolites and metabolized by the intestinal tract, and only about 15% of clopidogrel is Activated to generate active metabolites to exert their antiplatelet pharmacological effects. [0003] Clopidogrel is the basic antithrombotic drug for the treatment of acute coronary syndrome and percutaneous coronary intervention, but 4% to 30% of patients experience a decline in the efficacy of clopidogrel during treatment, and even clopidogrel resistance. Clopidogrel is a prodrug, which mainly depends on the metabolism of CYP2C19...

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Problems solved by technology

And on this basis, the probe is modified by locking nucleic acid and secondary structure, increasing the Tm value of the probe, and further improving the specificity of the probe binding. Finally, through the adjustment of the above system, the accuracy and specificity are evenly balanced. It can reach 100%, and the minimum detection concentration can reach 1ng / μL, which solves the problem of low accuracy and specificity in the detection of CYP2C19 gene polymorphism in the past

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