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Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP2D6 by using hybrid capture method

A technology of hybridization capture and genomics, which is applied in the field of a set of probes and library construction kits for detecting CYP2D6 polymorphisms of pharmacogenomics-related genes by hybridization capture method, which can solve the limitations of reduced activity of metabolic enzymes and detection of complex variant types Sex and other issues, to achieve the effect of short time-consuming, good clinical application prospects, and less time-consuming

Pending Publication Date: 2022-04-12
WUHAN ADICON CLINICAL LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CYP2D6*9 polymorphism is G2613-A2615 deletion, which can also cause decreased activity of metabolic enzymes
[0005] Traditional gene mutation detection methods, such as Sanger sequencing, pyrosequencing, and real-time fluorescent PCR, can only detect a single gene, or some exon mutations of a single gene, and have limitations in the detection of complex mutation types

Method used

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  • Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP2D6 by using hybrid capture method
  • Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP2D6 by using hybrid capture method
  • Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP2D6 by using hybrid capture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Implementation 1: Building the library

[0019] Enzyme digestion library construction

[0020] DNA fragmentation, end repair, and A-tailing (amplified regions)

[0021] First turn on the PCR instrument, and set the temperature of the thermal lid of the PCR instrument to 70°C.

[0022] 1.0.1 The initial amount of DNA library construction is 10 ~ 100ng, mixed well and then placed on ice for instant use.

[0023] 1.0.2 Go to the reagent preparation area, take a new 1.5ml centrifuge tube, and prepare the enzyme digestion reaction mixture. After thawing 10x Fragmentation Buffer and 5x Fragmentation Enzyme (Twist Bioscience) on ice, prepare a mixture according to the table below, mix well and centrifuge, then divide the mixture into eight tubes, and then transfer to the amplification area through the transfer window . (At the same time, the DNA purification magnetic bead solution used in the next step of purification can be taken out of the refrigerator, fully vortexed an...

Embodiment 2

[0111] Embodiment 2 clinical sample detection

[0112] One whole blood sample was taken from each of the four volunteers, and the polymorphism of the CYP2D6 gene in the four samples was detected. First extract the whole blood genomic DNA of 4 volunteers respectively, then build a library according to the method described in Example 1, and successfully construct the library (see figure 1 ), the library was sequenced on an Illumina high-throughput sequencer, and the sequencing parameter PE: 2×150. Then, after quality control of the disembarkation data, relevant databases such as PharmGKB, CPIC, etc. were used to conduct bioinformatics analysis on the data to obtain the distribution of CYP3A4 gene polymorphisms in each sample, and then to verify each site by next-generation sequencing. Rate 100%.

[0113] Volunteer 1 test results:

[0114]

[0115] Volunteer 2 test results:

[0116]

[0117] Volunteer 3 test results:

[0118]

[0119] Volunteer 4 test results:

[0...

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Abstract

The invention provides a group of probes and a kit containing the group of probes. The group of probes and the kit containing the group of probes are used for manufacturing a library building kit for detecting polymorphism of a pharmacogenomics related gene CYP2D6 by using a hybrid capture method. According to the present invention, the CYP2D6 genotype and the drug metabolism capability can be rapidly obtained through the one-time detection, and the kit has advantages of simple operation, low time consumption, easy automatic detection, large sample size related research, and great significance on the discovery of the new genotype.

Description

technical field [0001] The invention relates to a group of probes and a library construction kit for detecting the CYP2D6 polymorphism of pharmacogenomics-related genes by hybridization capture method. Background technique [0002] In recent years, with the gradual promotion of precision medicine, pharmacogenomics (pharmacogenomics, PGx) has developed rapidly as a powerful means to realize individualized drug delivery. PGx mainly studies gene polymorphisms that affect individual differences in drug reactions (including curative effect and adverse reactions), involving genes encoding drug metabolism enzymes, transporters, and targets. Changes in these gene characteristics will affect drug concentrations and targets in vivo Drug sensitivity of tissue. Therefore, the detection and analysis of these genomic biomarkers can help guide clinical precision medication, predict the occurrence of adverse drug reactions, and guide the development and application of new drugs. [0003] T...

Claims

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Application Information

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IPC IPC(8): C12N15/11C40B50/06C12Q1/6883C12Q1/6874
Inventor 周桂兰
Owner WUHAN ADICON CLINICAL LAB
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