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Deafness susceptibility gene screen test kit

A susceptibility gene and kit technology, which is applied in the field of kits for screening deafness susceptibility genes, can solve the problems of complicated operation, long operation time, and the extension cannot be continued.

Active Publication Date: 2010-03-31
GENERAL HOSPITAL OF PLA
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  • Abstract
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  • Application Information

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Problems solved by technology

The advantage is fast and high-throughput screening, but the operation is too complicated and the cost is too high, so it is not easy to popularize
[0013] 2. LDR method: with ligase detection reaction (LDR) as the core technology, there are currently kits for drug-induced deafness-related gene polymorphism sites (SNPs), which can detect multiple sites Mutation, but requires probe technology, hybridization, and interpretation of results after elution. There are many operating procedures and professional operations are required.
At the same time, although the cost of this method is lower than that of chip technology, it takes at least 5 hours for a detection operation
[0014] 3. RFLP method: Restriction fragment length polymorphism (RFLP) is the core technology. Currently, this type of kit is mainly composed of DNA extraction part, PCR part, and endonuclease reagent. The cost is lower than that of chip technology and LDR technology is relatively low, and the operation is relatively simple, but template preparation, PCR amplification, enzyme digestion and electrophoresis verification are required. The operation time is long, the process is cumbersome, and the relationship between each link is closely related. If one step is wrong, the next step cannot be carried out.
[0017] The four-primer ARMS-PCR method (Tetras primer amplification refractory mutationsystem-PCR, Tetras primer ARMS-PCR) belongs to the category of allele-specific PCR technology. , when the 3′ terminal base of the primer cannot be complementary to the template, the elongation efficiency is significantly reduced. Amplified bands of specific length

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Embodiment Construction

[0051] The invention is further described by means of the following examples.

[0052] The following examples are exemplary rather than limiting, and specific implementation methods can be determined according to the above technical solutions and reagent conditions.

[0053] 1. Selection of samples to be tested

[0054] The samples to be tested were selected from the "Huaxia Ten Thousand Newborn Hearing Rescue Project" undertaken by the People's Liberation Army General Hospital. All blood samples involved in this project were analyzed using the technical method of "DNA extraction-PCR amplification-sequencing" for the 1555 and 1494 sites of the mitochondrial 12S rRNA gene, the IVS7-2 site of the SLC26A4 gene, and the GJB2 gene. 235 loci were genotyped.

[0055] In this embodiment, the above-mentioned genotyped samples are selected as the samples to be tested, wherein 4 homozygous negative genotype samples, 4 heterozygous genotype samples, and 2 homozygous positive genotype sa...

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Abstract

The invention relates to a hereditable deafness susceptibility gene screen test kit. The kit uses mutation from C to T of 1,494 locus of a 12s rRNA gene, mutation from A to G of 1,555 locus of the 12srRNA gene, mutation from A to G of IVS7(-2) locus of an SLC26A4 gene and C(+ / -) of 235 locus of a GJB2 gene as detecting objects, designs and optimizes a set of specific primers respectively againsteach locus to be tested according to the PCR technical principle of a tetra-primer amplification refractory mutation system, amplifies the whole set of the primers in the same reaction tube, and performs primary multiple PCR amplification and primary gel electrophoresis on the four reaction tubes to obtain gene types of four loci simultaneously.

Description

technical field [0001] The invention belongs to the technical field of gene detection, in particular to a kit for screening deafness susceptibility genes. Background technique [0002] The key to the control and prevention of human diseases is to predict the disease, that is, to know the tendency of the disease, and then to carry out targeted health care. At present, there are five levels of disease inspection: tissue and organ level, preclinical level, cell level, protein level, and gene level (molecular level). From this we can see that genetic testing is the earliest early warning and the most accurate and highest level of diagnosis. Genetic testing is to accurately determine the physiological health status of relevant organs through the determination and location analysis of the subject's gene coding sequence, to detect the present and predict the future. International standardized operation, the results can be in line with international standards at any time. It enabl...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王秋菊魏宏泉
Owner GENERAL HOSPITAL OF PLA
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