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

Primer composition, kit and application for genetic detection of preimplantation hereditary deafness

A hereditary deafness and pre-embryo implantation technology, applied in the field of molecular biology, can solve the problems of high risk of misdiagnosis, long time-consuming, complicated operation, etc., to reduce the interference of results, improve accuracy, and reduce the risk of misdiagnosis

Active Publication Date: 2022-03-18
SUZHOU BASECARE MEDICAL DEVICE CO LTD
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Hereditary deafness is the main birth defect in my country, and the prognosis and treatment are difficult to achieve complete recovery. Therefore, tertiary prevention of deafness and birth defects has been carried out in China
However, traditional PGD detection mainly uses PCR-based next-generation sequencing technology or capillary electrophoresis to detect STR to complete the analysis. The detection throughput is low, the operation is complicated, and it takes a long time. It is usually impossible to realize the simultaneous detection and detection of pathogenic variants and polymorphic sites. analyze
This method can only detect a small number of loci, so the risk of misdiagnosis caused by allele dropout 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
  • Primer composition, kit and application for genetic detection of preimplantation hereditary deafness
  • Primer composition, kit and application for genetic detection of preimplantation hereditary deafness
  • Primer composition, kit and application for genetic detection of preimplantation hereditary deafness

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Analysis of pathogenic variants in family samples

[0060] 1. Sample preparation

[0061] The blood genomic DNA of both husband and wife and their affected offspring (proband) was extracted, the DNA concentration was measured by Qubit 2.0, and a certain amount was taken as the DNA sample to be tested.

[0062] 2. Target sequence capture and library construction

[0063] 1. Target Enrichment Capture

[0064] 1) After the DNA sample was vortexed and centrifuged, place it on an ice box, take two 0.2ml PCR tubes for each sample, and prepare the reaction system according to Table 3:

[0065] table 3

[0066] components Reaction volume (μl) dna sample 2(50ng) nuclease free water 4 5× Amplification Mixture 4

[0067] 2) After the preparation is completed, evenly distribute to two 0.2ml PCR tubes, 5μl in each tube. Add 5 μl of primer pool 1 to one tube, and 5 μl of primer pool 2 into the other tube, vortex to mix, and centrifuge...

Embodiment 2

[0101] Example 2 Pre-implantation detection of embryos

[0102] 1. Sample Preparation

[0103] Three to five cells from the trophectoderm of blastocyst-stage embryos were isolated as samples to be tested. The blastocyst stage embryos were sampled according to conventional embryo biopsy techniques, and the cells taken out were washed with 1×PBS (without Ca 2+ , Mg 2+ ) and washed 3 times, placed in a 0.2ml PCR tube with a volume not exceeding 2.5μl. Single-cell samples first use PicoPlex TM WGA kit for whole genome amplification.

[0104] 2. Target sequence capture and library construction

[0105] With embodiment 1.

[0106] The joints used for the 8 embryo samples to be tested G30218032601P01 to G30218032601P08 are SEQ ID NO.11 to SEQ ID NO.18 respectively.

[0107] 3. Fluorescent quantitative PCR to measure library concentration

[0108] With embodiment 1.

[0109] 4. On-machine sequencing and analysis of sequencing results

[0110] Dilute the library to 100pM, ta...

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 relates to a primer composition, a kit and an application for detecting preimplantation hereditary deafness genes. The present invention designs primer pairs for PCR amplification for the coding region sequences of the deafness-causing genes GJB2 and SLC26A4, and related SNP sites on chromosome 7 and / or 13, and can target the target region of the disease-causing gene through targeted capture technology. Enrichment, library construction, and subsequent sequencing and haplotype analysis can be further used to analyze related gene information, which has important guidance and research value for auxiliary diagnosis and treatment of hereditary deafness. The primer composition of the present invention can realize the simultaneous detection and analysis of pathogenic variants and polymorphic sites, so that the detection of pathogenic variants and linkage analysis can be completed in the same reaction system, reducing the interference of human factors on the results, and can improve The accuracy of detection can effectively reduce the risk of misdiagnosis caused by ADO.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a primer composition, kit and application for preimplantation hereditary deafness gene detection. Background technique [0002] Hereditary deafness is a large class of genetic diseases. According to WHO statistics, in 2013, the number of hearing disabled people in the world reached 360 million, accounting for 5.3% of the world's total population. Worldwide, 1 out of every 1,000 newborns is born with congenital deafness. children. According to statistics from various countries, congenital hearing impairment ranks first among birth defects, with an incidence rate of about 1‰~3‰. According to the results of the second national sample survey of disabled people released in December 2006, there are 27.8 million people with hearing and speech disabilities in my country, accounting for 33.52% of the total number of disabled people, and 900,000 new cases of birth defects each year, inclu...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 曹扬冒燕刘慧敏孔令印梁波
Owner SUZHOU BASECARE MEDICAL DEVICE CO LTD
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