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Detection kit for c.740G>T mutation of PJVK gene

A kit and gene technology, applied in the field of kits for detecting PJVK gene mutation genes, can solve rare problems and other problems

Inactive Publication Date: 2012-10-17
王秋菊 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the first discovery and reporting of the PJVK gene in 2006, scholars have found more than 10 mutations of the gene in patients from Iran, Morocco, Turkey and the Netherlands, but sensorineural deafness and auditory neuropathy / auditory neuropathy are rare in my country Mutations in this gene have been reported in patients with disorders

Method used

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  • Detection kit for c.740G>T mutation of PJVK gene
  • Detection kit for c.740G>T mutation of PJVK gene
  • Detection kit for c.740G>T mutation of PJVK gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] [Example 1] Extraction of blood sample to be tested and PCR amplification of PJVK gene coding region

[0063] 1. Preparation of blood sample DNA of the subject to be tested

[0064] 1. Research object

[0065] 150 patients with sporadic non-syndromic sensorineural deafness and auditory neuropathy / acoustic neuropathy spectrum disorder and 99 normal hearing controls without family history were screened for PJVK gene according to the following method. Among the 150 patients, 1 patient showed typical abnormal auditory function characteristics of auditory neuropathy spectrum disorder, and was diagnosed as non-syndromic auditory neuropathy spectrum disorder after clinical observation and systematic auditory function examination. The PJVK gene detection of this patient with auditory neuropathy spectrum disorder found that the patient was heterozygous for the c.740G>T mutation. No c.740G>T mutation was found in the screening of 99 normal hearing persons.

[0066] All partici...

Embodiment 2

[0096] [Example 2] Purification and Quantification of PCR Amplified Product of Coding Region of PJVK Gene

[0097] 1. Purification of PCR products——96-well plate method

[0098] 1. Add 50 μl sterile water to the 96-well plate containing the PCR product and mix well.

[0099] 2. Transfer it to the Millipore purification plate, put it on the vacuum pump for about 3 minutes, and see that there is no water in the purification plate.

[0100] 3. Add 50 μl of deionized water to the purification plate again, and continue to filter until there is no water in the purification plate.

[0101] 4. Remove the purification plate from the vacuum pump, add 20 μl of deionized water to the plate, let it rest for 15 minutes, shake it for another 15 minutes, and then suck it into a new 96-well plate.

[0102] 5. Store in a -20°C refrigerator.

[0103] 2. Quantification by electrophoresis

[0104] 1. Sample preparation

[0105] Take a 96-well spotting plate, add 6 μl of sample buffer to eac...

Embodiment 3

[0116] [Example 3] Direct Sequencing of PCR Amplified Product of Purified PJVK Gene Coding Region

[0117] 1. Purity and dosage requirements of PCR product DNA template

[0118] DNA purity: OD 260 / OD 280 =1.6~2.0.

[0119] DNA concentration: PCR product 10ng / μl.

[0120] DNA consumption:

[0121] PCR product

[0122]

[0123] 2. Sequencing reaction

[0124] 1. The reagents required for the sequencing reaction should be freshly prepared, and the reagents that need to be sterilized by autoclaving must be sterilized before use. The equipment required for the sequencing reaction (such as 384-well plates, tips, etc.) should also be clean and sterile.

[0125] 2. In order to ensure the freshness of sequencing samples and reaction reagents, it should be operated on ice when adding samples.

[0126] 3. The current reaction system is 5 μl, and the amount of various reagents added is shown in Table 2.

[0127] Table 2 Sequencing reaction system of PJVK gene PCR amplificat...

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Abstract

The invention provides a detection kit for c.740G>T mutation of the PJVK gene. Causes and types of sensorineural deafness and auditory neuropathy / auditory neuropathy spectrum disorders are diagnosed through detecting whether patients have the mutation gene or not. The mutation gene and a detection method mentioned above are beneficial for clinical screening of PJVK mutation for patients with sensorineural deafness and auditory neuropathy / auditory neuropathy spectrum disorders, providing bases for diagnosis and treatment of patients with sensorineural deafness and auditory neuropathy / auditory neuropathy spectrum disorders.

Description

technical field [0001] The present invention relates to the field of gene detection, in particular to a kit for detecting PJVK gene mutation gene; the present invention also relates to PJVK mutation gene and its use in the diagnosis and / or treatment of sensorineural deafness and auditory neuropathy / auditory neuropathy spectrum disorder application. Background technique [0002] Auditory Neuropathy Spectrum Disorders (ANSD) is a hearing disorder in which the outer hair cells of the cochlea function normally, but the sound signal cannot be transmitted synchronously from the inner ear to the brain. It is an abnormality in the process of auditory information transmission and processing . The audiological examination showed hearing loss mainly in low-frequency hearing loss, the auditory brainstem response could not be elicited, and the cochlear microphonic potential and otoacoustic emissions were mostly normal. This type of disease was defined and named in the 1990s. It is a ne...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王秋菊
Owner 王秋菊
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