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Gene chip for detecting functional central nerve damage and its production method

A central nervous system and gene chip technology, applied in the field of genes, can solve the problems of high cost and large amount of data, and achieve the effect of simple research work, speed up and progress

Inactive Publication Date: 2007-08-01
陈云 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional high-throughput gene expression profiling chips are costly and require a lot of data. Many gene expression data are unnecessary or useless data for researchers.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Preparation of Gene Chip for Diagnosis and Detection of Functional Central Nervous Injury

[0041] The construction of a.DNA square matrix: 381 kinds of specific genes and negative controls (bacterial control probes or human housekeeping genes (Housekeeping Genes)) closely related to central nervous system damage listed in Table 1 are randomly classified according to their functions Arrangement (in the computer detection program of the chip, there is a list of 381 gene probes generated according to the random arrangement sequence of Oligo probes on the spotting machine) on the DNA-ReadyType II Slide glass slide produced by Clontech Company in the United States. The slide has been processed by combinatorial chemistry, which not only provides efficient nucleic acid binding capacity, but also prevents cross-contamination between spots to the greatest extent.

[0042] b. Probe design and in situ synthesis: According to two important parameters of the gene chip: d...

Embodiment 2

[0044] Example 2: Application of gene chip for diagnosis and detection of functional central nervous system injury

[0045] a. RNA extraction: Use Trizol reagent, chloroform, isopropanol, and 75% alcohol to extract RNA from serum, cerebrospinal fluid, or damaged tissue. After repeated centrifugation, filtration, and washing, the obtained RNA is stored at 4°C until use.

[0046] b. Reverse transcription into cDNA: After mixing RNA with amine-modified random primers (such as 2-hydrogen adenine), RNase inhibitors, aa-dUTP / dNTPs, DTT and SSII Reverse Transcriptase for about 2 hours, use EDTA (pH8 .0) Stop the reaction and then hydrolyze the RNA with NaOH. The reverse-transcribed cDNA was extracted by MinElute filter column produced in the United States, and the purified cDNA was obtained after repeated centrifugation, filtration, washing and drying.

[0047] c. Fluorescent labeling: the sample cDNA was labeled with a fluorescent marker (Cy5 or Cy3) using a Qia-quick filter column...

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PUM

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Abstract

The invention relates to gene chip used for detecting functional central nervous damage diagnosis and the preparing method. said gene chip is equipped with oligonucleotide gene probe array, the oligonucleotide biomolecule of said probe array is the special biological gene factor playing improtant role in multiple central nervous system damage process simutaneously, relating to netural cell death and tissue necrosis, inflammation, irritated response, ion dysequilibrium inside cell, signal transmission, abnormal control of cell cycle, abnormal cell construction and nerve degeneration damage morbid physiology and molecular biology active gene, respectively. The gene chip can be used to check the conditioning way, expression level and change rule of biological factor relative to cerebral ischemia damage inside body in the beginning of clinic for high risk group of cerebral apoplexy, and to predict possibility of cerebral apoplexy; and used to conditioning way and change rule of special biological factor relative to nerve damage inside nerve cell after central nervous system damage, and relationship between cell death and disease occurancy, and provides checking criterion for cilinical treatment.

Description

technical field [0001] The invention belongs to the field of gene technology, and relates to a gene chip for the diagnosis and detection of functional central nervous system damage and a manufacturing method thereof, in particular to a specific detection and central nervous system damage (including brain damage caused by cerebral apoplexy or cerebrovascular accident) , traumatic brain injury and spinal cord injury) pathophysiological and molecular biological changes closely related to the oligonucleotide chip of the main neurotrauma shared genes. Background technique [0002] Central nervous system injury (including brain injury caused by cerebral apoplexy or cerebrovascular accident, traumatic brain injury and spinal cord injury) is a major disease that threatens human health. It results in severe disability and an unusually high mortality rate, as well as enormous medical rehabilitation costs. Every year in the United States, about 430,000 people suffer from cerebral apop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈云刘谦黄卫宫岩华
Owner 陈云
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