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Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP3A4 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 CYP3A4 polymorphisms of pharmacogenomics-related genes by hybridization capture method, which can solve the problems of false positive or false negative and detection sensitivity Low cost, complicated operation and other problems, to achieve the effect of simple operation, short time consumption and high efficiency

Pending Publication Date: 2022-06-24
南昌艾迪康医学检验实验室有限公司
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Currently commonly used methods for detecting CYP3A4 gene polymorphisms include DNA sequencing analysis (DNA sequencing analysis), restriction fragment length polymorphism polymerase chain reaction (Polymerase chain reaction-restriction fragment length polymorphism, PCR-RFLP), high performance liquid chromatography (Highper- formanceliquidchromatography, HPLC), capillary electrophoresis (Capillary Electrophoresis, CE), etc., these methods all have disadvantages such as complicated operation, long time-consuming, high cost, low detection sensitivity and easy to cause false positive or false negative

Method used

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  • Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP3A4 by using hybrid capture method
  • Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP3A4 by using hybrid capture method
  • Group of probes and library building kit for detecting polymorphism of pharmacogenomics related gene CYP3A4 by using hybrid capture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Implementation 1: Construction of the library

[0020] Enzymatic digestion library construction

[0021] DNA Breaking, End Repair and A-tailing (Amplified Region)

[0022] First turn on the thermal cycler and set the thermal lid temperature of the thermal cycler to 70°C.

[0023] 1.0.1 The initial amount of DNA library construction is 10-100 ng, mix well and leave on ice for use.

[0024] 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 the mixture according to the following table, mix well and centrifuge, then dispense the mixture into eight tubes, and then transfer it to the amplification area through the transfer window . (At the same time, the DNA purification magnetic bead solution used in the next step can be taken out of the refrigerator, fully shaken and mixed, and then transferred...

Embodiment 2

[0129] Example 2 Clinical sample detection

[0130] One whole blood sample from each of the four volunteers was taken to detect the CYP3A4 gene polymorphism in these four samples. First, the whole blood genomic DNAs of 4 volunteers were extracted respectively, and then the library was constructed according to the method described in Example 1, and the library was successfully constructed (see figure 1 ), the library was sequenced on an Illumina high-throughput sequencer, and the sequencing parameter PE: 2×150. Then, after the quality control of the off-machine data, relevant databases such as PharmGKB, CPIC, etc. are used to perform bioinformatics analysis on the data, and the CYP3A4 gene polymorphism distribution of each sample is obtained. rate 100%.

[0131] Volunteer 1 test results:

[0132]

[0133] Volunteer 2 test results:

[0134]

[0135] Volunteer 3 test results:

[0136]

[0137] Volunteer 4 test results:

[0138]

[0139] It can be seen from the d...

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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 CYP3A4 by utilizing a hybrid capture method. According to the present invention, the genotype and the drug metabolism capability of CYP3A4 can be rapidly obtained through one-time detection, and the kit has advantages of simple operation, low time consumption and easy automatic detection, is especially suitable for the large sample size related research, and provides great significance for the discovery of the new genotype.

Description

technical field [0001] The invention relates to a set of probes and a library-building kit for detecting the polymorphism of a pharmacogenomics-related gene CYP3A4 by using a hybridization capture method. Background technique [0002] In recent years, with the gradual promotion of precision medicine, 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 responses (including efficacy and adverse reactions), involving genes encoding drug metabolizing enzymes, transporters, targets, etc. Changes in these gene characteristics will affect drug concentration and target in vivo. Therefore, the detection and analysis of these genomic biomarkers can help guide clinical precision medicine, predict the occurrence of adverse drug reactions, and guide the development and application of new drugs. [0003] The CYP450 family is a family of isoenzymes involve...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6883C40B50/06C12Q1/6874
CPCC12Q1/6883C40B50/06C12Q1/6874C12Q2600/106C12Q2600/156C12Q2531/113C12Q2535/122
Inventor 周桂兰
Owner 南昌艾迪康医学检验实验室有限公司
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