Method for quickly detecting CYP2C19 genetic polymorphism and kit

Abstract

The invention relates to the field of molecular biology, discloses a method for quickly detecting CYP2C19 genetic polymorphism and a kit, and provides the method for quickly detecting the CYP2C19 genetic polymorphism and the kit. A sample to be tested is mixed with sample treating fluid, a DNA extraction step is not needed, and simple and easy treatment on samples can be realized in short time; moreover, further through a selective amplification primer and a selective detection probe, effective distinguishing between wild type variants and mutant type variants of the CYP2C19 gene can be realized, and a whole process from treatment of the samples to acquisition of a final detection result is completed within 1 hour; moreover, the accuracy, the specificity and the sensitivity of the detection can be ensured.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method and kit for rapidly detecting CYP2C19 gene polymorphism. Background technique [0002] Cytochrome P450 (CYP or P450) is one of the main metabolic enzymes in the liver. CYP2C19 is an important member of the second family of P450 enzymes, involved in about 12% of the P450 enzymes responsible for metabolizing drugs, and is an important drug-metabolizing enzyme in the human body. At present, many single nucleotide polymorphisms of CYP2C19 have been found, causing the occurrence of CYP2C19 alleles, among which CYP2C19*2 (G681A, rs4244285), CYP2C19*3 (G636A, rs4986893) and CYP2C19*17 are common in Asian populations allelic type. CYP2C19*2 and CYP2C19*3 can cause the loss of the enzyme activity encoded by the CYP2C19 gene, weaken the ability to metabolize the substrate, and increase the blood drug concentration, thereby causing adverse drug reactions related to the ...

AI Technical Summary

Problems solved by technology

[0005] The main disadvantages of the sequencing method are: (1) Before the PCR reaction, it is necessary to extract DNA from the sample, check the quality and concentration, the operation is complicated, and there is a risk of confusing the sample

(2) When detecting multiple sites, one sample needs to perform multiple reactions, the detection cost is high, the demand for the amount of template is high, the operation intensity is high, and it is easy to cause operation errors and pollution

The main limiting factors of this method are: a) if the mutated site is a weakly mismatched base sequence, the ARMS primers cannot effectively distinguish the wild-type variant and the mutant variant of the target nucleic acid sequence, so that the selectivity of the method is limited. effects, which can lead to false-negative or false-positive results

b) ARMS-PCR only contains a selective amplification mechanism but does not have selective detection, and cannot perform genotype-specific detection. Therefore, when selective amplification is wrong, it will lead to completely wrong conclusions, that is, false negatives and false positives. result

c) ARMS-PCR primer design must cover polymorphic site regions, when these regions have special sequences or structures, amplification (efficiency, specificity, selectivity) will be affected

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