Screening method of infective gene capture probe related to dementia, capture genome, product and application

A technology for capturing probes and pathogenic genes, which is applied in biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection.

Inactive Publication Date: 2020-01-14
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection methods in the prior art are for the detection of a specific type of the above-mentioned types of dementia, and cannot effectively detect the above-mentioned common types at the same time

Method used

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  • Screening method of infective gene capture probe related to dementia, capture genome, product and application
  • Screening method of infective gene capture probe related to dementia, capture genome, product and application
  • Screening method of infective gene capture probe related to dementia, capture genome, product and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Preparation of capture probes for human dementia-related genes

[0087] This example is aimed at the relevant pathogenic genes of Alzheimer's disease, frontotemporal dementia, Parkinson's disease-related dementia, amyotrophic lateral sclerosis-related dementia and dementia with Lewy bodies: APP, PSEN1, PSEN2, APOE, MAPT, SNCA, FUS, TARDBP, CHCHD10, CHMP2B and TBK1, according to the following design rules of the capture probe set, the capture probes for capturing human dementia-related genes were obtained:

[0088] (a) The length of the probe is 121bp;

[0089] (b) probe design to merge adjacent regions;

[0090] (c) The probe design interval extends 20bp from the target interval to the two wings;

[0091] (d) The probe reference is the sense strand of the genome;

[0092] (e) The consensus sequences on both wings of the probe are shown in SEQ ID NO.1 and SEQ ID NO.2;

[0093]

[0094]

[0095] (f) Ensure probe specificity by comparing whole genome in...

Embodiment 2

[0104] Example 2 Specific capture and sequencing

[0105] This embodiment uses the Alzheimer's disease, frontotemporal lobe dementia, Parkinson's disease-related dementia, amyotrophic lateral sclerosis-related dementia and Lewy body dementia-related gene capture probe set provided in Example 1, and based on the liquid phase Hybrid capture technology enriches target sequences. The technical route of liquid-phase hybridization capture is: 1) preparing a hybridization probe library, 2) using the probes to enrich the target gene, and 3) sequencing the enriched DNA sequence with a high-throughput sequencer. The use of liquid phase hybridization technology can effectively enrich the target gene, reduce the requirement of sample size, and better control the cost of sequencing.

[0106] The library construction method comprises the following steps:

[0107] 1. DNA extraction and fragmentation

[0108] DNA was extracted from 6 peripheral blood samples using the QIAamp DNA Blood Mini...

Embodiment 3

[0151] Example 3 Alzheimer's disease, frontotemporal dementia, Parkinson's disease-related dementia, amyotrophic lateral sclerosis-related dementia and Lewy body dementia-related gene screening

[0152] This example provides a method for screening genes related to Alzheimer's disease, frontotemporal dementia, Parkinson's disease-related dementia, amyotrophic lateral sclerosis-related dementia, and dementia with Lewy bodies. Genetic screening for Alzheimer's disease, frontotemporal dementia, Parkinson's disease-related dementia, amyotrophic lateral sclerosis-related dementia, and Lewy body dementia. The library prepared in Example 2 was subjected to paired-end sequencing on the Illumina sequencing platform. At the same time, the tag sequence on the sample is also sequenced. Analyze the source of sequencing data through data analysis and perform data analysis. The data were compared with the UCSChg19 database through BWA. The results are shown in the table below:

[0153] ...

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Abstract

The invention relates to the technical field of biology, and particularly provides a screening method of an infective gene capture probe related to dementia, a capture genome, a product and an application. The method provides a screening condition of the capture probe. The screening condition is optimized, and the method specifically comprises restriction on the related condition of selection of atarget section, the length of the capture probe, requirements on specificity, GC content, the number of probes and the like. The infective gene capture probe related to dementia comprises coding regions of 11 genes, infective genes for detecting five kinds of diseases of Alzheimer's disease, frontotemporal dementia, dementia related to Parkinson's diseases, dementia related to amyotrophic lateralsclerosis and lewy body dementia can be realized, and reference can be provided for molecular genetics diagnosis of ill individuals, screening of familial Alzheimer's diseases, frontotemporal dementia, the dementia related to Parkinson's diseases, the dementia related to amyotrophic lateral sclerosis and the lewy body dementia, and corresponding heredity consultation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a screening method for capturing probes for dementia-related pathogenic genes, capturing genomes, products and applications. Background technique [0002] With the aging of society, the incidence of dementia is increasing year by year. There are many types of dementia, the main types include Alzheimer's disease (AD), frontotemporal dementia (FTD), Parkinson's disease-related dementia (PDD), muscle Atrophic lateral sclerosis-related dementia (ALS-FTD) and Lewy body dementia (LBD). The clinical diagnosis of this group of different types of dementia mainly relies on auxiliary means such as clinical manifestations, imaging, blood and cerebrospinal fluid related indicators. So far, there is no specific and sensitive indicator to distinguish the above dementia types. Clinical diagnosis is difficult, especially in the early stage of the disease . Genetic factors play an important role in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/6883
CPCC12Q1/6883C12Q2600/156
Inventor 吴志英陶青青
Owner ZHEJIANG UNIV
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