Primer combination for detecting allele CYP2C19*3 and detection kit thereof

Abstract

The invention provides a primer combination for detecting an allele CYP2C19*3. The primer combination includes an upstream primer p1, a downstream primer p2, a gene extension primer p3 and a gene extension primer p4 respectively having the sequences shown in Seq ID No.1-4. The invention also provides a primer combination-containing kit for detecting the allele CYP2C19*3. Based on design thoughts of specific primer extension and a high-resolution melting curve, genotyping can be directly carried out in a premise of not carrying out special amplification post-processing. The method perform genotyping by using a fluorescent dye identification extension product, requires no use of special labeled probes, and is simple in method, low in cost, and easy to popularize in clinic.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a primer combination and a detection kit for detecting allele CYP2C19*3. Background technique [0002] There are multiple alleles in the CYP2C19 gene, including *1, *2, *3...*17, etc. Among them, the allele with normal enzyme activity is CYP2C19*1, while the enzymes encoded by CYP2C19*2 and CYP2C19*3 alleles are inactive. The slow metabolism of drugs caused by CYP2C19*2 and CYP2C19*3 accounts for 99% of the slow metabolism of Orientals, so the detection technology of CYP2C19*2 and CYP2C19*3 has been paid more and more attention. If the patient is *1 / *1, it is a fast metabolizer; if it is *1 / *2 or *1 / *3, it is an intermediate metabolizer; if it is *2*2 or *2 / *3 or *3 / * 3 belongs to the slow metabolism type. The CYP2C19*2 allele can be identified by detecting the G681A variation of CYP2C19, and the CYP2C19*3 allele can be identified by detecting the G636A variation of CYP2C19. ...

AI Technical Summary

Problems solved by technology

Therefore, the specificity is much lower than that of sequencing methods

[0012] 4. PCR-LDR method: This method involves two steps of gene-specific amplification and ligase detection reaction, which are costly and time-consuming, and because special treatment after amplification is required, it is easy to cause sample contamination

[0013] 5. Solid-phase chip method: This method involves multiple steps such as gene-specific amplification, hybridization, and detection. It is costly and time-consuming, and the processing after amplification is relatively complicated, which may easily cause sample contamination.

The disadvantage of this method is that the gene-specific primers must terminate at the G636A site, resulting in the inability to optimize the primer design, resulting in affected amplification efficiency

Therefore, in the case of a small amount or low quality of sample DNA, it is likely to cause failure of amplification and typing

Another disadvantage of this method is the need to use specially labeled probes, which increases the cost

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