Primer pair used for detecting CYP2C19 genotypes, fluorescent probe, reagent kit and method

Abstract

The invention discloses a reagent kit used for detecting CYP2C19 genotypes. Allelic genotypes of CYP2C19*2(681G / A) and CYP2C19*3(636G / A) of a CYP2C19 gene are detected by designing a specific primer pair and a fluorescent probe. Asymmetric PCR and the fluorescent probe melting curve analysis technology are combined, different genotypes are judged through melting peaks of specific temperatures, detection of six genotypes related to CYP2C19*2 and CYP2C19*3 can be achieved in one-tube PCR, subsequent processing is not needed, the possibility of reactant pollution is avoided to the largest extent, operation is easy, consumed time is short, specificity is good, accuracy is high, and results are easy to interpret.

Description

technical field [0001] The invention relates to the field of allele typing, in particular to a primer pair, fluorescent probe, kit and method for detecting CYP2C19 gene CYP2C19*2 and CYP2C19*3 genotyping. Background technique [0002] With the development of medical technology, new drugs emerge in endlessly, but there are more and more medication problems. Many drugs used clinically, such as inhibitors-omeprazole, antidepressants-fluoxetine, antiepileptic drugs-phenytoin Tumor drug-cyclophosphamide, etc., there are individual differences in drug use and drug metabolism. For example, the same drug with the same dosage is used in different patients, but the curative effect may be completely different. Some are cured, while others may have serious adverse drug reactions. Even death, according to the statistics of the World Health Organization, about 19 million people die from adverse drug reactions in the world every year. [0003] Studies have shown that individual difference...

AI Technical Summary

Problems solved by technology

[0007] At present, there are many methods for the detection of CYP2C19 genotyping. The conventional methods include: (1) direct sequencing method, by extracting human peripheral venous blood, extracting genomic DNA, and performing DNA direct sequencing after amplification to determine the genotype ; This method can directly display the position and type of known mutation sites, however, its operation steps are cumbersome, reagents and instruments are expensive, the detection period is long and the cost is high

(2) Restriction sequence analysis, based on the principle of disappearance or increase of the restriction endonuclease cutting site formed by mutation; although this method is simple and economical, it is difficult to achieve high-throughput parallel analysis, heterozygous

(3) Chip method, using a known nucleotide sequence as a probe to hybridize with a labeled target nucleotide sequence, and performing qualitative and quantitative analysis through signal detection; this method can be used for high-throughput analysis, however, Its technical cost is high, the detection steps are complicated, and the analysis pan is narrow

The above methods are difficult to meet the needs of clinical testing for CYP2C19 genotyping detection

Images