Kit for detecting single gene mutations of 50 hereditary diseases, and probe group used by kit

A single-gene mutation and kit technology, applied in the medical field, can solve the problems of huge data volume, small throughput, and complex genetic information analysis.

Active Publication Date: 2021-02-02
北京迈基诺基因科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the next-generation sequencing technology (Nextgenerationsequencing, NGS), traditional genetic disease genetic diagnosis technologies (such as Sanger sequencing, fluorescence in situ hybridization, comparative genomic hybridization, gene chip, etc.) have the disadvantages of low throughput, slow speed, and high precision. low cost, high cost
However, NGS itself also has the problems of huge data volume and complex analysis of genetic information.

Method used

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  • Kit for detecting single gene mutations of 50 hereditary diseases, and probe group used by kit
  • Kit for detecting single gene mutations of 50 hereditary diseases, and probe group used by kit
  • Kit for detecting single gene mutations of 50 hereditary diseases, and probe group used by kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0201] Example 1. Preparation of kits for detection of single-gene mutations in 50 genetic diseases

[0202] 1. Preparation of probe set

[0203]1. Design and synthesize 1059 probes based on the nucleotide sequences of the mutated genes involved in 50 single-gene genetic diseases. Each probe consists of 108 nucleotides, including DNA fragment 1, DNA fragment 2 and DNA fragment 3 from the 5' end to the 3' end. The nucleotide sequence of DNA fragment 1 is 5'-GACTACATGGGACAT-3'. The nucleotide sequence of DNA fragment 3 is 5'-GGAACCTACGACGTA-3'. Both DNA fragments 2 consist of 78 nucleotides. DNA segment 2 is part of the mutated gene involved in 50 monogenic diseases.

[0204] The principles of probe design are as follows: obtain the exon sequences of the mutated genes involved in 50 single-gene genetic diseases from the UCSC database, extend them by 50 bp before and after, and extend to 50 bp if they are less than 50 bp after the extension, and extract the reference sequence...

Embodiment 2

[0221] Embodiment 2, the establishment of the method for detecting whether there is a genetic disease single gene mutation by the kit prepared in embodiment 1

[0222] A. The kit prepared in Example 1 establishes a method for non-invasive prenatal detection of whether the fetus has 45 kinds of genetic diseases and single gene mutations

[0223] After a lot of experiments, the inventors of the present invention have established a method for non-invasive prenatal detection of whether the fetus has 45 kinds of genetic disease single gene mutations using the kit prepared in Example 1. Specific steps are as follows:

[0224] 1. Obtain the type of mutation site, mutation frequency and sequencing depth of the 45 mutation genes of the pregnant woman to be tested (see Table 1, except for CYP21A2, SMN1, HBA1, HBA2, and MT-RNR1 genes)

[0225] (1) Extraction of genomic DNA from pregnant women to be tested

[0226] 10 mL of peripheral blood of pregnant women to be tested was collected w...

Embodiment 3

[0350] The beneficial effect of the kit prepared by embodiment 3, embodiment 1

[0351] Sample R-001 and sample R-003, sample R-002 and sample R-004 are the same normal human peripheral blood sample, and the data volume of sample R-001 and sample R-003 is similar. Sample R-002 and sample R-004 have a similar amount of data.

[0352] Samples R-001 and R-002 were taken, and a target sequence capture library was constructed using a universal BL-blocking kit (a kit from Mikino), and then high-throughput sequencing and data analysis were performed.

[0353] Take samples R-003 and R-004, use the kit prepared in Example 1 to construct a target sequence capture library, and then perform high-throughput sequencing and data analysis.

[0354] The analysis results are shown in Table 3 (Precise BL is the kit prepared in Example 1). The results showed that, compared with the general BL-blocking kit, the capture efficiency and average sequencing depth of the kit prepared in Example 1 were...

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Abstract

The invention discloses a kit for detecting single gene mutations of 50 hereditary diseases, and a probe group used by the kit. The probe group consists of 1059 specific probes, and nucleotide sequences of the 1059 specific probes are sequentially shown as SEQ ID NO:1 to SEQ ID NO:1059. The inventors of the invention develop the kit based on the liquid phase capture technology through a large number of experiments, and on the premise of ensuring the gene screening accuracy, the sequencing cost is greatly reduced, the sequencing depth is improved, and genetic variation information of specific regions of 50 single-gene hereditary diseases are more accurately found. The kit has important application value.

Description

technical field [0001] The invention belongs to the medical field, and in particular relates to a kit for detecting single-gene mutations of 50 genetic diseases and a probe set used therefor. Background technique [0002] Genetic diseases refer to diseases caused by changes in genetic material (such as gene mutations or chromosomal abnormalities). So far, more than 4,000 human single-gene genetic diseases have been identified, and the total number of diseases is huge [0003] Studies have shown that the average person carries 2.8 pathogenic mutations of genetic diseases. Most single-gene genetic diseases are recessive, and healthy parents may carry the same pathogenic mutations. Some routine prenatal examinations (such as imaging tests, nuclear type analysis, etc.) cannot judge the condition of the fetus, so carrier screening is carried out before pregnancy or in the first trimester to screen out the pathogenic mutation sites carried by the parents, prevent birth defects in...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6811C12N15/11C40B40/06
CPCC12Q1/6883C12Q1/6811C40B40/06C12Q2600/156C12Q2535/122C12Q2537/165
Inventor 伍建姬晓雯王海丽
Owner 北京迈基诺基因科技股份有限公司
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