Gene chip for non-invasive prenatal diagnosis of high-risk hereditary hearing loss and preparation method

A technology for hereditary deafness and prenatal diagnosis, applied in the field of genetic diagnosis, can solve the problems of high-throughput, large-scale gene polymorphism analysis, difficult to meet clinical application, complicated operation, etc., and achieve high repeatability and short time , the effect of simple operation steps

Inactive Publication Date: 2012-10-10
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional detection methods such as restriction fragment length polymorphism (Restriction Fragment Length Polymorphism, RFLP), single strand conformational polymorphism (Single Stranded Conformational Polymorphism, SSCP), disguised high performance liquid chromatography (Denaturing High Performance Liquid Chromatography, DHPLC), mass spectrometry (Mass Spectrometry, MS) and other complex operations, high cost, poor accuracy, and high-throughput, large-scale genetic polymorphism analysis cannot be performed, which is difficult to meet the requirements of clinical applications

Method used

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  • Gene chip for non-invasive prenatal diagnosis of high-risk hereditary hearing loss and preparation method
  • Gene chip for non-invasive prenatal diagnosis of high-risk hereditary hearing loss and preparation method
  • Gene chip for non-invasive prenatal diagnosis of high-risk hereditary hearing loss and preparation method

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Embodiment 1

[0028] Example 1. Preparation of gene chip for non-invasive prenatal diagnosis of high-risk hereditary deafness

[0029] This embodiment relates to a gene chip for non-invasive prenatal diagnosis of high-risk hereditary deafness. The preparation method of the gene chip includes the following steps:

[0030] Wash the slides with double distilled water, soak them overnight in lye, take them out, wash them three times with double distilled water, then soak them in HCl with a volume fraction of 1% for 30 minutes, and put them in a slide tank after cleaning. use. The cleaned slides were aminated with 4-aminobutyltriethoxysilane solution, and then treated with isothiocyanate with phenylisothiocyanate solution, dried with nitrogen, and placed in a 4°C place to avoid Light preservation;

[0031] The chip Zip probe is a random combination of oligonucleotide fragments with a length of 24bp. Use the BLAST function on the NCBI website to compare the homology of these oligonucleotide seq...

Embodiment 2

[0033] Example 2. Non-invasive prenatal diagnosis of high-risk hereditary deafness using gene chips

[0034] 2.1 Sample preparation

[0035] Plasma samples were prepared, and free DNA in plasma was extracted with QIAamp Circulating Nucleic Acid Kit (Qiagen, Basel, Switzerland).

[0036] 2.2 Selection of mutation detection sites

[0037] The 8 mutation sites and probe core sequences that need to be detected are shown in Table 1.

[0038] Table 1

[0039]

[0040] "del" in the mutant form means a deletion mutation, such as 35delG means the deletion of the 35th base G in the nucleotide sequence shown in SEQ ID NO.25; ">" means a substitution mutation, such as 707T>C means SEQ ID NO. The 707th base in the nucleotide sequence shown in .26 is mutated from T to C. By analogy, the positions represented by the first 5 of the above 8 mutations are in the nucleotide sequence shown in SEQ NO ID.25:

[0041] Whether the 35th base is mutated, and the 35th base G is missing;

[0042...

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Abstract

The invention relates to a gene chip for non-invasive prenatal diagnosis of high-risk hereditary hearing loss and a preparation method in the technical field of gene diagnosis. The gene chip comprises a substrate and probes fixed on the substrate, wherein the probes are nucleotide sequences shown as SEQ ID NO.1-24. Usage of the gene chip includes the steps of sample preparation, multiplex PCR (polymerase chain reaction) amplification, ligase detection reaction, chip hybridization, chip scanning and result obtaining. Beside, the invention further relates to a reagent kit comprising the gene chip. The gene chip for diagnosis is simple in operation steps, only requires the multiplex PCR reaction, the ligase detection reaction and hybrid scanning once, is high in detection specificity and good in stability so as to be capable of correctly distinguishing homozygotes from heterozygotes of each locus and high in repeatability of repeated experiments, is short in detection time and low in cost as the substrate can be universally used for other chips by means of universal chip technology.

Description

technical field [0001] The invention belongs to the technical field of gene diagnosis, and specifically relates to a gene chip for non-invasive prenatal diagnosis of high-risk hereditary deafness and a method for using it. Background technique [0002] Deafness is one of the most common disabling diseases and clinical genetic diseases. The reported incidence of newborns is as high as 1 to 3‰. Studies have found that about 60% of newborns' deafness is related to genetic factors, that is, hereditary deafness. About 70% of them are inherited in an autosomal recessive manner. Hereditary deafness is caused by mutations in a single gene or compound mutations in different genes. More than 120 deafness-related genes have been discovered so far, with high genetic heterogeneity of genes and loci. In recent years, studies have found that GJB2, SLC26A4 and other genes are the most common deafness-related genes, 235delC, 299delAT, 176del16 on GJB2, and IVS7-2A>G, 2168A>G on SLC26...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C40B40/06
Inventor 秦胜营段涛贺林霍燃孟梦
Owner SHANGHAI JIAO TONG UNIV
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