Primer combination for differentiating verapamil individualized medication types

Abstract

The invention provides a primer combination for differentiating verapamil individualized medication types. An extension primer which has different molecular weights at different single nucleotide polymorphism (SNP) sites is utilized, so that through matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), a plurality of sites pertinent to metabolism of a hypertension reducing medicine namely verapamil can be detected at the same time, and finally the primer combination for differentiating verapamil individualized medication types is obtained. A preparation method of the primer combination comprises the steps of according to 8 objective SNP sites to be detected, respectively designing a multiplex amplification primer and an extension primer; compounding amultiplex amplification primer reaction system and an extension primer reaction system; in the reaction systems, performing amplification and a single-base extension reaction on the 8 objective SNP sites at the same time with multiple sets of primers; and performing time-of-flight mass spectrometry analysis on products after the single-base extension reaction, identifying the genotypes of different drug metabolism pertinent SNPs according to products of different molecular weight extension primers represented by mass spectrum peaks, and guiding the medication of the hypertension reducing medicine namely the verapamil. The primer combination disclosed by the invention can detect 8 metabolism pertinent SNP sites of the hypertension reducing medicine namely the verapamil at the same time, and has the advantages of being low in cost, free from synthetized probes, short in time consumption, simple and convenient in result analysis, and extremely broad in application field.

Description

technical field [0001] The invention belongs to the field of molecular biology detection, and relates to a method for detecting multiple PCR single-base extension products by using mass spectrometry characteristic peaks and its products. The method can simultaneously detect multiple PCR single-base extensions in four multiplex PCR reactions Amplified oligonucleotide products. More specifically, the method uses different time-of-flight mass spectrometry characteristic peaks generated by different target oligonucleotide fragments in the process of mass spectrometry typing, and simultaneously detects multiple target SNP sites to guide the antihypertensive drug Vera The use of Pami. Background technique [0002] Human genetic information is stored in the genome. In 2002, the Human Genome Project, an international cooperation project, was finally completed, drawing a fine map of the human genome structure and providing a reference sequence for related research. The human genome...

AI Technical Summary

Problems solved by technology

However, the detection method and kit provided by the invention patent only target the polymorphic site of the adrenergic receptor gene, and the detection objects have limitations

Moreover, this method mainly uses the detection method of fluorescent quantitative PCR. Since fluorescent quantitative PCR needs to design specific probes for the variation of SNP sites, the throughput of this method is low, and only one polymorphic site can be measured in one experiment. Suitable for multiple SNP detection

In addition, if it is necessary to obtain all relevant SNP information, it is necessary to conduct multiple detection tests, which will increase the cost

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