Drug metabolic enzyme related gene SNP fluorescence labeling composite amplification kit

Abstract

The invention discloses a drug metabolic enzyme related gene SNP fluorescence labeling composite amplification kit. The kit comprises a mixture of specificity non-labeled primers and share labeled primers of five SNP loci, performs detection by combining PCR composite amplification with a capillary electrophoresis technology and can simultaneously detect genotypes of the five SNP loci of a drug metabolic enzyme related gene in a single tube within 3 hours. By applying the kit, the drug metabolic enzyme related gene is detected, the relation of the adverse drug reaction and genetic variation is researched, and a theoretical support is provided for rational drug use.

Description

technical field [0001] The invention belongs to the technical field of biological in vitro diagnosis, and in particular relates to a drug-metabolizing enzyme-related gene SNP fluorescent label composite amplification kit and a use method thereof. Background technique [0002] Cytochrome P450 enzymes (cytochrome P450, CYP), also known as drug-metabolizing enzymes, mainly exist in liver microsomes and play a very important role in the biotransformation of exogenous compounds (including drugs and poisons) and endogenous substances. Its activity determines the metabolic rate of the drug, which is directly related to the clearance rate of the drug, and is the first phase enzyme of drug metabolism. The genes of P450 enzymes mainly include three families, CYP1, CYP2, and CYP3, among which CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4, and CYP2E1 are the most important subtypes, which account for more than 80% of the total amount of CYP450 in the liver. More than 90% of the drugs metabol...

AI Technical Summary

Problems solved by technology

Among them, the biggest disadvantage of the fluorescent quantitative PCR method and the sequencing method is the occurrence of false negatives and false positives, and both need to detect the sites one by one. When there are many sites, the operation is complicated and the cost is high.

The SnaPshot detection method also has the following disadvantages: for example, the digestive enzymes used for partial purification are expensive, the operation steps are cumbersome, and the workload is heavy, and the detection success rate of sites with unknown mutations in the primer binding region is low, etc.

Five simultaneous reactions are required to obtain the genotypes of five SNP sites, and this method has disadvantages such as high cost of synthetic probes and inability to perform multiple detections

Chinese patent application number 201310433222.8, "a kit and method for detecting CYP3A4 gene polymorphism", specifically related to gene sequencing technology, although this method only needs one PCR amplification, the sequencing steps are cumbersome and time-consuming

Chinese patent application number 20130137910.X, "Primer system for detecting polymorphic sites related to human cytochrome P450 and its application", which involves multiplex PCR technology and single base extension technology, which can target Seven polymorphic sites of different genes are detected simultaneously, but this method has disadvantages such as expensive reagents and complicated operation process

After searching the existing technical documents, it is found that there is no report on the detection of CYP450-related gene polymorphisms using this method

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