Gene panel kit of Immotile cilia syndrome and application of gene panel kit

A syndrome and kit technology, applied in the field of biomedicine, can solve the problems of slow detection speed, inability to detect a large number of genes at the same time, and high cost, and achieve the effect of improving the depth of sequencing

Pending Publication Date: 2019-11-12
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Finally, traditional genetic detection methods cover a small number of genes, have a low detection throughput, and cannot detect a large number of genes at the same time. The detection speed is slow, the efficiency is low, and the cost is high. Such as Sanger sequencing, etc., usually only for the inspection of some sites

Method used

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  • Gene panel kit of Immotile cilia syndrome and application of gene panel kit
  • Gene panel kit of Immotile cilia syndrome and application of gene panel kit
  • Gene panel kit of Immotile cilia syndrome and application of gene panel kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Genomic DNA sample extraction, preparation

[0040] Use QIAamp DNA blood mini kit (QIAGEN, Cat: Y5-51106) to extract and purify DNA from venous blood of subjects, the specific steps are as follows:

[0041] 1. Take venous blood, centrifuge at 1000g for 15 minutes at room temperature, absorb 1ml of blood cells, add 3ml of DNA lysate, mix thoroughly with a vortex shaker, and mark the name and date.

[0042] 2. Pipette 20 μl proteinase K to the bottom of a 1.5ml centrifuge tube.

[0043] Add 200μl blood sample to the centrifuge tube, then add 200μl Buffer AL to the sample, vortex for 15s to mix.

[0044] 3. After incubating at 56°C for 10 minutes, centrifuge quickly to remove the droplets remaining in the cap of the 1.5ml centrifuge tube.

[0045] 4. Add 200 μl of absolute ethanol, and vortex for 15 seconds to mix. After shaking, quickly centrifuge, transfer the mixture to a QIAamp Mini centrifugal column, and centrifuge at 6000g for 1min.

[0046] 5. Place t...

Embodiment 2

[0052] Example 2 Target gene whole exome capture, library construction

[0053] Design and synthesize the probe sequence, as shown in SEQ ID NO.1 to SEQ ID NO.49, and labeled with biotin, prepare the gene Panel liquid phase capture probe.

[0054] The sample obtained in Example 1 was captured and detected by the whole exon gene panel liquid phase capture probe system of the present invention, and the target region library result captured before sequencing was obtained.

[0055] The gene panel liquid chip system of the present invention is used to efficiently enrich the DNA of the whole exon region of the target gene DNA in Example 1, and then perform high-throughput and high-depth sequencing on the Illumina Hiseq platform. The gene panel liquid-phase capture probe system of the present invention is used for library construction and capture experiments. The selected supporting reagents and consumables are provided by corresponding suppliers, including but not limited to the rea...

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Abstract

The invention relates to the field of biomedical technologies and disease diagnosis, and particularly discloses a gene panel capture kit for detecting genetic mutation of a pathogenic gene of Immotilecilia syndrome and application of the gene panel capture kit. The liquid phase capture kit of the gene panel comprises 49 probes corresponding to the related pathogenic gene of the Immotile cilia syndrome. The design of the gene panel capture kitsynthesizes the pathogenic factors of the Immotile cilia syndrome genetic defects of cilia immobility syndrome in multiple layers and aspects, comprehensive covering is achieved, the target capture of the gene panel capture kit has obvious pertinence, and the pathogenic gene with large flux can be accurately and simultaneously captured and genetic-detected. Human target gene exon region DNA is efficiently enriched, and then high-throughput and high-depth sequencing is conducted on a second generation sequencing platform, the depth and coverage ofgene sequencing can be effectively improved, and convenience is provided for obtaining more accurate genetic mutation information of gene pathogenicity and disease prevention and diagnosis.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and specifically provides a design and application of a gene panel (Panel) capture kit for detecting multiple genes and multiple sites of ciliary immobility syndrome. Background technique [0002] Primary ciliary dyskinesia (PCD) is an autosomal recessive genetic disease caused by abnormal ciliary function due to movement disorders caused by structural defects of cilia, with an incidence of about 1 / 10000 to 1 / 20000. The clinical manifestations of PCD patients are repeated respiratory infections, sinusitis, bronchiectasis, visceral transposition, infertility, etc., which seriously affect the lives of patients. Kartagener syndrome (KTS) is a type of primary ciliary movement disorder, and its clinical phenotype also includes visceral transposition. The cilia structure in the human body is closely related to the following tissues or organs and their functions: such as the respiratory tract, Eust...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/156
Inventor 乐雍建傅应云黄启俊刘盛国
Owner SHENZHEN PEOPLES HOSPITAL
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