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High-throughout sequencing method of genes relative to neurological and psychiatric drugs and application of high-throughout sequencing method

A high-throughput and spiritual technology, applied in the field of neuropsychiatric gene detection, can solve the problems of unfavorable data accumulation and scientific research development, low flexibility and high cost, and achieve the advantages of scientific research and clinical use, improve detection sensitivity, Wide coverage effect

Active Publication Date: 2018-07-31
广州海思医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although gene chips can achieve high throughput, they are expensive, inflexible, and rely on known gene polymorphisms, which have limitations in discovering new mutation sites, which is not conducive to the accumulation of data and the development of scientific research
The development of next-generation sequencing has made whole-genome sequencing of different individuals a reality, which can comprehensively mine the genetic variation at the DNA level of different individuals, but high cost is the bottleneck restricting its wide application

Method used

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  • High-throughout sequencing method of genes relative to neurological and psychiatric drugs and application of high-throughout sequencing method
  • High-throughout sequencing method of genes relative to neurological and psychiatric drugs and application of high-throughout sequencing method
  • High-throughout sequencing method of genes relative to neurological and psychiatric drugs and application of high-throughout sequencing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Detection of neuropsychiatric drug-related genes based on the Illumina sequencing platform

[0045] Reagents: Multiplex PCR Kit (Vazyme), KOD-Plus-0902 (TOYOBO), TIANamp BloodDNA Kit (TIANGEN)

[0046] 1. Genomic DNA acquisition:

[0047] Three psychiatric patients: Sample1, Sample2, Sample3 anticoagulant blood samples were extracted using Tiangen Blood DNA Extraction Kit, and were extracted according to the kit instructions. The obtained genomic DNA was detected by NanoDrop 2000 ultra-micro ultraviolet spectrophotometer for A260 / A280, and Qubit quantified the sample concentration C (ng / μl). The result is as follows:

[0048] sample

Sample1

Sample2

Sample3

A260 / A280

1.82

1.90

1.86

C (ng / μl)

31

29

38

[0049] 2. The first round of multiplex PCR reaction

[0050] Human genomic DNA is mixed with PCR primers and PCR amplification reagents for PCR reaction to amplify the target gene. The system is carr...

Embodiment 2

[0104] The anticoagulant blood samples of three psychiatric patients: Sample1, Sample2, and Sample3 were taken for detection. The operation steps were performed according to Example 1. After sequencing and analysis on the Illumina sequencing platform, the following results were obtained:

[0105]

[0106]

[0107] (Note: W means wild type, M means homozygous mutant, H means heterozygous mutant. Human beings are diploid organisms with two homologous chromosomes. Wild type means that both homologous chromosomes are homologous at this site. No mutation; homozygous mutant means that the mutation is present on both chromosomes; heterozygous mutant means that one chromosome is mutated and the other is not.)

[0108] For the three patients, if the attending physician intends to use the drug risperidone, and the metabolism of risperidone is mainly related to the CYP2D6 enzyme, according to the above results, it can be known that the Sample1 CYP2D6 genotype is a normal wild type, ...

Embodiment 3

[0110] Three psychiatric patients: Sample1, Sample2, and Sample3 were tested for anticoagulant blood samples and sequenced using the Sanger method. The sequencing results were compared with those of the patent of the present invention. The results are shown in the following table:

[0111]

[0112]

[0113] It can be seen from the above table that this method is completely consistent with the results obtained by the Sanger sequencing method, which is the gold standard for sequencing. However, the Sanger sequencing method has fewer detection sites and a longer detection time; compared with whole genome resequencing, this method (target region capture) has a lower sequencing volume, which reduces the sequencing cost and is highly targeted, with deeper coverage and data accuracy higher.

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Abstract

The invention belongs to the field of neurological and psychiatric gene detection and particularly relates to a high-throughout sequencing method of genes relative to neurological and psychiatric drugs and application of the high-throughout sequencing method. The invention provides the high-throughout sequencing method of the genes relative to the neurological and psychiatric drugs and the application of the high-throughout sequencing method. The method comprises the following steps of acquisition of a genome DNA, target gene multiplex PCR amplification reaction and product purification, linker sequence PCR reaction and product purification, library detection, high-throughout sequencing and the like. The method can be used for detecting 29 SNP loci of 20 genes relative to drugs for neurological and psychiatric diseases at one time, can cover a vast majority of first-line and second-line drugs for resisting schizophrenia, depression, anxiety, epilepsy, Parkinson's disease, hyperkineticdisorder, pains and the like and provides guidance to medication of a patient to the maximum.

Description

technical field [0001] The invention belongs to the field of neuropsychiatric gene detection, and in particular relates to a high-throughput sequencing method and application of neuropsychiatric drug-related genes. Background technique [0002] At present, major changes have taken place in the spectrum of human diseases. Neuropsychiatric diseases are becoming more and more advanced in the spectrum of human diseases, and their incidence is still rising with the increasing work and life pressure that people are facing. At present, mental illness accounts for about 20.8% of the total disease burden in my country, and it is expected to increase to 25% by 2020. In June 2009, The Lancet published the results of an epidemiological survey of mental disorders in four provinces in China. According to the survey, about 173 million people in China suffer from different types of mental disorders. As the aging population continues to increase, the incidence and prevalence of neurological...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12Q1/6883
CPCC12Q1/6869C12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16C12Q2537/143C12Q2535/122C12Q2525/191
Inventor 周煌凯钟诗龙邓美英陆嫚云徐毓璇姚啟聪
Owner 广州海思医疗科技有限公司
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