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Rapid, simple and convenient deafness gene detection method and kit

A kit and primer combination technology is applied in the field of biomedicine to achieve the effects of saving detection time, high precision and short detection time period

Inactive Publication Date: 2021-07-13
北京尔惠基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method has been widely used in the detection of antigens and antibodies, but it is rarely used in the field of nucleic acid detection

Method used

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  • Rapid, simple and convenient deafness gene detection method and kit
  • Rapid, simple and convenient deafness gene detection method and kit
  • Rapid, simple and convenient deafness gene detection method and kit

Examples

Experimental program
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Effect test

Embodiment 1

[0034]Embodiment 1, the primer combination of detection deafness gene mutation and the preparation of deafness gene PCR-colloidal gold chromatography kit

[0035] This embodiment provides a combination of primers for detecting the mutation of the deafness gene, which can be used to detect the c.235delC mutation of the GJB2 gene (recorded as GJB2:c.235delC, that is, the deletion of the 235th C in the coding region of the wild-type GJB2 gene) and the mutation of the SLC26A4 gene. The c.919-2A>G mutation (denoted as SLC26A4:c.919-2A>G, that is, the A mutation at position 919 in the coding region of the wild-type SLC26A4 gene is changed to G), which is detected by the combination of primers for detecting GJB2:c.235delC and Composition of primer sets to detect SLC26A4:c.919-2A>G.

[0036] The primer combination for detecting GJB2:c.235delC consists of G235 WF, G235 MF and G235 WR, and G235 WF, G235 MF and G235 WR are marked by the 5' end of the single-stranded DNA shown in sequence...

Embodiment 2

[0060] Example 2, Deafness Gene PCR-Colloidal Gold Chromatography Kit Detection Sample

[0061] 1 sample

[0062] Sample to be tested: peripheral blood sample.

[0063] 2 sample testing

[0064] Utilize the deafness gene PCR-colloidal gold chromatography kit of embodiment 1 to detect the sample to be tested, the steps are as follows:

[0065] 2.1 Sample processing

[0066] Take 200 μL of the sample to be tested, add 100 μL of sample release agent, let it stand on ice for 10 minutes, and centrifuge at 10,000 g for 2 minutes. Take 100 μL of the supernatant for use (storage at -80°C for a long time).

[0067] 2.2PCR amplification

[0068] Configure the PCR amplification reaction solution (20 μL for each reaction) as shown in the table below, wherein the concentrations of the six primers in the detection solution A are all optimal concentrations, that is, 200 nM:

[0069] components 1 reaction volume Detection solution A 7.5μL Detection solution B 1...

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Abstract

The invention discloses a rapid, simple and convenient deafness gene detection method and a kit. The kit comprises a primer combination for detecting deafness gene mutation, the primer combination is composed of G235WF, G235MF, G235WR, SIV72WF, SIV72WR and SIV72MR, and the sequences of the primer combination are as follows: the 5'tail ends of the G235WF, the G235MF, the G235WR, the SIV72WF, the SIV72WR and the SIV72MR are respectively marked with fluorescein FITC, TAMATA, biotin biotin, biotin biotin, fluorescein FAM and DIG. Experiments prove that the kit disclosed by the invention can be used for rapidly, accurately and sensitively detecting mutation conditions of two sites of GJB2: c.235delC and SLC26A4:c.919-2A>G.

Description

technical field [0001] The invention relates to a quick and simple deafness gene detection method and kit in the field of biomedicine. Background technique [0002] Deafness is one of the most common diseases that threaten human health and disability, and it is also the most common disease that causes communication barriers. Deafness is divided into non-synthetic deafness and comprehensive deafness according to different performance characteristics. Studies have shown that at least 50% of congenital deafness is caused by genetic factors, and about 77% of non-synthetic deafness is autosomal recessive. There are many pathogenic factors for deafness. Among all deaf patients, hereditary deafness accounts for about 50%; deafness-causing gene mutations can be found in 70%-80% of hereditary deafness patients. [0003] According to different ways of inheritance, hereditary deafness is divided into five types: autosomal dominant inheritance (AD), autosomal recessive inheritance (AR)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11G01N33/543G01N33/558
CPCC12Q1/6883C12Q1/686G01N33/54313G01N33/558C12Q2600/156
Inventor 韩菲
Owner 北京尔惠基因科技有限公司
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