Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Probe set and kit for detecting alpha thalassemia and beta thalassemia related pathogenic genes

A technology for thalassemia and disease-causing genes, which is applied in the direction of recombinant DNA technology, biochemical equipment and methods, and microbial measurement/inspection, can solve the problems of inconvenient detection and high cost of comprehensive detection

Active Publication Date: 2021-02-12
北京迈基诺基因科技股份有限公司
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinical gene screening for thalassemia generally uses Gap-PCR and PCR-RDB methods to detect three deletions and three point mutations of the common HBA1 / HBA2 genes, and 17 point mutations of the HBB gene. The gene kit products are all separate products, which is inconvenient for testing and the cost of comprehensive testing is relatively high

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Probe set and kit for detecting alpha thalassemia and beta thalassemia related pathogenic genes
  • Probe set and kit for detecting alpha thalassemia and beta thalassemia related pathogenic genes
  • Probe set and kit for detecting alpha thalassemia and beta thalassemia related pathogenic genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Preparation of complete set of reagents and kits for detection of thalassemia-related pathogenic genes

[0075] 1. A complete set of reagents for detecting thalassemia-related pathogenic genes

[0076] The complete set of reagents for detecting thalassemia-related pathogenic genes provided in this example, according to the full-length sequences of HBA1, HBA2, and HBB genes in the Hg19 reference genome database as the target sequence, refer to the method in patent WO2013 / 003585: along each sequence in sequence Stack past design probes. The length of each probe sequence used in the present invention is 78bp. The probe synthesized above was biotin-labeled to form the capture probe of this example.

[0077] The specific operation is as follows: Synthesize the above-mentioned probes by a method known in the art, uniformly mix them in a total volume of 1.2ml of dH2O, take 15ul of them and use general-purpose PCR primers (the sequence at the 5' end is GACTACATGGGAC...

Embodiment 2

[0106] Embodiment 2 detects the method for thalassemia

[0107]

[0108]

[0109] Table 2: Rare variants in thalassemia genes

[0110]

[0111]

[0112] The method for detecting thalassemia-related pathogenic genes using the complete set of reagents and kits in Example 1 of the present invention is as follows:

[0113] 1. Whole genome library preparation

[0114] After fragmenting the gDNA to 150-250bp fragments, use the KAPA Library Construction Kit (Catalog No. KK8504) to construct a genome-wide library according to the instructions.

[0115] 2. Capture library preparation of thalassemia-related pathogenic gene probe sets

[0116] Capture using the probe set and capture kit described in Example 1, the specific steps are as follows:

[0117] 1) Take 1 μg of the genome-wide library in step 1, add 13 μl of enrichment buffer and 5 μl of a complete set of reagent solution (this solution is obtained by dissolving the set of reagents in Example 1 with water, and the ...

Embodiment 3

[0143] The corresponding beneficial effect of the optimized probe design method of embodiment 3

[0144] The probe group 1 designed by the probe design method for optimizing gc content described in the present invention adjusts the probe density formula according to the GC content:

[0145] D. go =a+a*10*|gc-0.5|

[0146] where D pc Represents the probe density of a certain GC content, a represents the probe density when the GC content is 50%, which is 20. gc stands for GC content,

[0147] With the probe set 2 designed without gc optimization, after using the methods described in Example 1 and Example 2 for probe preparation, capture sequencing, and bioinformatics analysis, as shown in Table 1, figure 1 , figure 2 As shown, the results of the basic statistics and the uniformity of target region coverage show that: the uniformity of the capture coverage region of probe set 1 designed by the optimized probe design method is better, and more proportion of Targeted regions...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a probe set for detecting alpha thalassemia and beta thalassemia related pathogenic genes. The probe set comprises a first probe set for capturing and detecting HBA1 and HBA2 and a second probe set for capturing and detecting HBB, wherein the first probe set and the second probe set are designed to refer to the Hg38 / Hg19 version of a human genome and are oligonucleotide probes, and the coverage area of the first probe set is chr16:147743-185701 of the Hg38 version. According to the Hg19 version of chr16: 197742-235700, the Hg38 version of chr11: 5225597-5227178 and theHg19 version of chr11: 5246827-5248408, the invention further provides a kit which comprises the first probe set, the second probe set, an enrichment buffer solution, 0.1 M of a hybridization buffer solution, 1 M of a binding buffer solution, a first rinsing solution and a second rinsing solution. The probe set provided by the invention can simultaneously detect HBA1, HBA2 and HBB genes, and can detect 323 mutation types of alpha thalassemia and beta thalassemia which are known at present.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a probe set and a kit for detecting α-thalassemia and β-thalassemia-related pathogenic genes. Background technique [0002] Thalassemia is a high-incidence autosomal recessive genetic disease that seriously threatens human health. The global carrier rate is about 2%, and the carrier rate in southern my country is 3%-24%. [0003] Thalassemia is a hereditary hemolytic hemoglobinopathy in which peptide chain synthesis is reduced or absent due to a defect in the globin gene. The most common are alpha thalassemia and beta thalassemia. Clinically, according to the clinical manifestations, it can be divided into light type (thalassemia carrier), intermediate type and heavy type. [0004] Adult hemoglobin is a tetramer composed of two types of 4 hemoglobin monomers (a pair of α-peptide chains and a pair of β-peptide chains). Only tetramers composed of two α-peptide chains and two β-pept...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/6883C12Q1/6869C12N15/11
CPCC12Q1/6883C12Q1/6869C12Q2600/156C12Q2535/122C12Q2565/507C12Q2563/131C12Q2523/308
Inventor 伍建姬晓雯王海丽刘娜韩路
Owner 北京迈基诺基因科技股份有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com