Method for detecting copy number of CYP2D6 gene based on unlabeled probe melting technology, primer pair and unlabeled probe combination and kit

Abstract

The invention relates to a method for detecting the copy number of a CYP2D6 gene, in particular to a method for detecting the copy number of the CYP2D6 gene based on an unlabeled probe melting technology, a primer pair and unlabeled probe combination and a kit. The defects of high cost, long period, low universality and the like of a traditional gene copy number detection method are overcome. The method comprises the following steps: (1) designing and synthesizing a non-labeled probe, a biotin labeled downstream primer and a biotin labeled upstream primer; (2) obtaining genome DNA of a sample to be detected; (3) preparing a PCR reaction system; (4) carrying out PCR reaction; (5) acquiring a single-chain PCR (Polymerase Chain Reaction) product by utilizing the magnetic beads marked by streptavidin; and (6) melting curve analysis and CYP2D6 gene copy number determination. The CYP2D6 gene copy number analysis can be completed on the premise that a fluorescent probe is not needed.

Description

technical field [0001] The invention relates to a method for detecting the copy number of CYP2D6 gene, in particular to a method for detecting the copy number of CYP2D6 gene based on unlabeled probe melting technology, a combination of primer pairs and unlabeled probe and a kit. Background technique [0002] The product encoded by the human CYP2D6 gene is a phase I drug-metabolizing enzyme involved in the metabolism of about 30% of clinical drugs. There are extensive gene polymorphisms and copy number variations in the CYP2D6 gene, and copy number variations can cause significant changes in the enzyme activity. By analyzing the copy number and polymorphism of CYP2D6 gene, the rapid detection of the enzyme activity can be realized. [0003] There are various traditional methods for detecting gene copy number, including TaqMan fluorescent probe technology, multiplex ligation-dependent probe amplification (MLPA) and next-generation sequencing technology. Although these techno...

AI Technical Summary

Problems solved by technology

Although these technologies are widely used, their shortcomings are still obvious: TaqMan fluorescent probes are expensive to synthesize raw materials, difficult to obtain, and usually cannot overcome the defects of low signal-to-noise ratio, and poor repeatability of experimental data

The operation of MLPA technology is complicated, and capillary electrophoresis is required for product analysis, and the whole experiment takes too long

The next-generation sequencing technology is suitable for the detection of large samples, but the sequencing platform it relies on is expensive and requires professional experimental operators and data analysis personnel, so it is not suitable for general laboratories, primary hospitals or medical laboratories

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