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Primer and probe for screening human deafness multi-gene mutation and application method thereof

A human and probe technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as poor repeatability and cross-contamination

Inactive Publication Date: 2014-06-11
曾骥孟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gene chip technology is used for the detection of genetic deafness-causing genes, which has the advantages of high throughput and high degree of automation, but has disadvantages such as poor repeatability, easy cross-contamination, and false positive results.

Method used

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  • Primer and probe for screening human deafness multi-gene mutation and application method thereof
  • Primer and probe for screening human deafness multi-gene mutation and application method thereof
  • Primer and probe for screening human deafness multi-gene mutation and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0175] In this embodiment, the detection of the GJB2 gene is taken as an example to illustrate the method of single-tube fluorescence PCR detection of the sample to be tested.

[0176] The samples used in the experiment were a total of 1,200 samples of deaf-mute students and their family members, including 400 deaf-mute students and 800 family members.

[0177] Utilize above-mentioned fluorescent PCR method to identify the method for GJB2 gene mutation in 1200 people as follows:

[0178] (1) DNA extraction of samples:

[0179] A 400 μL whole blood sample was drawn from each person, and DNA was extracted using RBC MagCore Genomic DNA Whole Blood Kit kit (Cat.No.: MGB400-04) according to the kit's operating instructions. After the quality of the extracted DNA was detected by an ultraviolet spectrophotometer, the extracted DNA was adjusted to 10 ng / μL with Tris-HCl solution (10 mmol / L, pH 8.0) as a template for PCR amplification;

[0180] (2) Fluorescent PCR amplification:

[...

Embodiment 2

[0203] In this example, the detection of the GJB3 gene is taken as an example to illustrate the method of single-tube fluorescent PCR detection of the sample to be tested.

[0204] The samples used in the experiment were a total of 1,200 samples of deaf-mute students and their family members, including 400 deaf-mute students and 800 family members.

[0205] Utilize above-mentioned fluorescent PCR method to identify the method for GJB3 gene mutation in 1200 people as follows:

[0206] (1) DNA extraction of samples:

[0207] A 400 μL whole blood sample was drawn from each person, and DNA was extracted using RBC MagCore Genomic DNA Whole Blood Kit kit (Cat.No.: MGB400-04) according to the kit's operating instructions. After the quality of the extracted DNA was detected by an ultraviolet spectrophotometer, the extracted DNA was adjusted to 10 ng / μL with Tris-HCl solution (10 mmol / L, pH 8.0) as a template for PCR amplification;

[0208] (2) Fluorescent PCR amplification:

[0209...

Embodiment 3

[0228] In this embodiment, the detection of the SLC26A4 gene is taken as an example to illustrate the method of single-tube fluorescence PCR detection of the sample to be tested.

[0229] The samples used in the experiment were a total of 1,200 samples of deaf-mute students and their family members, including 400 deaf-mute students and 800 family members.

[0230] The method for identifying the SLC26A4 gene mutation in 1200 people using the above fluorescent PCR method is as follows:

[0231] (1) DNA extraction of samples:

[0232] A 400 μL whole blood sample was drawn from each person, and DNA was extracted using RBC MagCore Genomic DNA Whole Blood Kit kit (Cat.No.: MGB400-04) according to the kit's operating instructions. After detecting the amount of extracted DNA with a UV spectrophotometer, adjust the extracted DNA to 10 ng / μL with Tris-HCl solution (10 mmol / L, pH 8.0) as a template for PCR amplification;

[0233] (2) Fluorescent PCR amplification:

[0234] The PCR sys...

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Abstract

The invention discloses a primer and a probe for screening human deafness multi-gene mutation and an application method thereof, and belongs to the technical field of gene detection. The invention discloses a primer and a probe for screening human deafness GJB2 gene mutation GJB2-M1, GJB-M2 or GJM2-M3, GJB-M4 and GJB2-M5, and an application method thereof, a primer and a probe for screening human deafness GJB3 gene mutation GJB3-M1 or GJB3-M2 and an application method thereof, and primer and a probe for screening human deafness SLC26A4 gene mutation SLC26A4-M1 and SLC26A4-M2 and an application method thereof. By adopting the primer and the probe and the application method thereof provided by the invention, a plurality of samples can be detected in the same polymerase chain reaction (PCR) reaction, and compared with the traditional detection method, the application method has the advantages of being high in accuracy rate, fewer in steps, and less in elapsed time.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to primers, probes and application methods for multi-gene mutation screening of human deafness. Background technique [0002] Deafness is a common disease that seriously affects human health. It is also one of the most common clinical genetic diseases. Because of its complex etiology, high incidence and difficult treatment, it greatly affects the patient's interpersonal communication and quality of life. According to statistics, the incidence of deafness in newborns is about 1‰ to 3‰, and about 70 million people in the world suffer from different degrees of hearing loss. my country is the country with the largest population of hearing impairments in the world. Among people, hearing disabilities account for about 24% of the total, involving tens of millions of families across the country. [0003] There are many causes of deafness, including genetic factors and env...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2561/113C12Q2537/143C12Q2563/107
Inventor 曾骥孟庄建立刘斌赖金娇
Owner 曾骥孟
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