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Method for detecting hereditary hearing loss relative connexin 26 gene GJB2 mutation and kit for detection

A technology of connexin and hereditary deafness, which is applied in the field of genetic engineering, can solve problems such as the inability to comprehensively and accurately analyze the relationship between deafness and the inability to accurately determine hereditary deafness, and achieve the effect of facilitating diagnosis and improving the detection rate

Inactive Publication Date: 2009-02-11
ドングァン アオマイヤ ジェネティック テクノロジー カンパニー リミテッド
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the detection of GJB2 gene mutation is limited to the sequence determination of exon 2 in the coding region or restriction endonuclease analysis for a single common mutation site. Although this detection and analysis method can detect whether the GJB2 gene is mutated, However, it is only limited to the sequence determination of the coding region of the GJB2 gene, that is, exon 2, or the restriction enzyme analysis of a single common mutation site, so it is impossible to comprehensively and accurately analyze the relationship between deafness and GJB2 gene mutation, and therefore cannot accurately determine Whether it is hereditary deafness

Method used

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  • Method for detecting hereditary hearing loss relative connexin 26 gene GJB2 mutation and kit for detection
  • Method for detecting hereditary hearing loss relative connexin 26 gene GJB2 mutation and kit for detection
  • Method for detecting hereditary hearing loss relative connexin 26 gene GJB2 mutation and kit for detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Detection of the GJB2 mutation of the connexin 26 gene in an autosomal recessive family with hereditary deafness

[0024] 1. Test samples

[0025] One deaf family with autosomal recessive inheritance as the transmission characteristic was selected, with a total of 27 people, including 3 deaf patients. Among the subjects, there were 2 deaf patients, and peripheral whole blood DNA was extracted from the subjects (Yuan Huijun et al., Chinese Journal of Otorhinolaryngology, 1998, 33(2): 67-70; Li Weimin et al., Journal of Clinical Otorhinolaryngology, 2001, 15 (Supplement): 53-58), as a test sample. For the pedigree of the family, see figure 1 .

[0026] The proband, male, 15 years old, Han nationality, had a full-term vaginal delivery. He was deaf when he was 1 year old, and his medication history was unknown. Physical examination: the general condition is normal, the tympanic membrane is intact, and the temporal bone CT examination excludes inner ear deform...

Embodiment 2

[0088] Example 2 Detection of GJB2 mutation in sporadic cases of hereditary deafness-related connexin 26 gene of unexplained sensorineural deafness

[0089] 1. Test samples

[0090] Select 50 sporadic patients with unexplained sensorineural deafness, and extract peripheral whole blood DNA from the individuals (Yuan Huijun et al., Chinese Journal of Otorhinolaryngology, 1998, 33(2): 67-70; Li Weimin et al., Clinical Otorhinolaryngology Department Journal, 2001, 15 (Supplement): 53-58), as a test sample.

[0091] 2. Primer design 3. PCR amplification reaction 4. Purification and sequence determination of PCR products are the same as in Example 1.

[0092] 5. Results: See Table 1.

[0093] Table 1 Example 2 GJB2 gene detection results

[0094] genotype number of people 9G>A / IVS1+1G>A 1 235delC / 257C>G 1 235delC / 299delAT 2 235delC / wide type 1 235delC / 235delC 1 299delAT / 299delAT 1 176del16 / 139G>T 1 wide type / wide type 42...

Embodiment 3

[0095] Example 3 Type I kit for GJB2 mutation of hereditary deafness-associated connexin 26 gene and its application

[0096] 1. The type I kit (100 copies) suitable for heel blood spots contains the following components:

[0097] (1) Solution I (the main component is 5% Chelex) 25ml;

[0098] (2) PCR amplification reaction reagents, including dNTP, 2×PCR buffer (for exon 1 amplification), 10×PCR buffer (for exon 2 amplification), Mg++, triple distilled water, Taq enzyme 1 (for amplifying exon 1), Taq enzyme 2 (for amplifying exon 2);

[0099] (3) the forward primer F1 of the amplification GJB2 gene basic promoter region, exon 1 and its cut region described in claim 2 and the amplification GJB2 gene basic promoter region, exon 1 and its Reverse primer R1 for the cut region, forward primer F2 for amplifying exon 2 and its cut region, and reverse primer R2 for amplifying exon 2 and its cut region mixed in equal proportions;

[0100] (4) Instruction manual.

[0101] 2. Detect...

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Abstract

The invention relates to a method for testing gap linking protein 26 gene GJB2 mutation related to hereditary hearing impairment, which completely enlarges GJB2 gene base boot sector, exon 1, exon 2 and shear zone by polymerase chain reaction; and then, DNA sequence is measured to detect whether GJB2 genetic mutation exists; the method of the invention can completely cover all the genetic mutation of the GJB2 gene base boot sector, the exon 1, the exon 2 and the shear zone, so that detectable rate of the GJB2 genetic mutation and diagnosis rate of hereditary hearing impairment related to the GJB2 gene are improved; compared with that sequence measurement is separately carried out on a GJB2 code area, the method is more comprehensive and reliable, so as to be beneficial to the diagnosis of hereditary hearing impairment.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a detection method for the GJB2 mutation of the connexin 26 gene associated with hereditary deafness. The invention also relates to a kit for detecting the GJB2 mutation of the hereditary deafness-related gap connexin 26 gene in vitro according to the method. Background technique [0002] Deafness is one of the most common genetic diseases clinically, and it is the most common disease that causes speech communication impairment. According to statistics from various countries, 1 / 2000-1 / 1000 children are born with severe deafness; at the same time, more than half of childhood deafness is caused by genetic factors (Marazita ML, et al., Am J Med Genet.1993, 46: 486-491; Kalatzis V et al., Hum Mol Genet. 1998, 7:1589-1597; Morton CC, Hum Mol Genet. 2002, 11:1229-1240). 70% of hereditary deafness is nonsyndromic hearing impairment (NSHI), and the remaining 30% is syndromi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 袁永一戴朴韩东一金政策
Owner ドングァン アオマイヤ ジェネティック テクノロジー カンパニー リミテッド
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