Detection type gene chip for detecting various peptitis

A gene chip and hepatitis technology, which is applied in the field of gene chips for detecting hepatitis mutations, can solve the problems that it is difficult to replace conventional detection methods, the chip detection cost is high, and the detection cost is high, so as to facilitate large-scale promotion and application and reduce the amount of samples to be tested. The effect of less, high degree of automation

Inactive Publication Date: 2003-01-22
赵伟 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] (2) Existing chips can usually only diagnose one disease, and the cost of chip detection is relatively expensive, so it is difficult to replace conventional detection methods at present
[0022] Hepatitis virus is a rout

Method used

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  • Detection type gene chip for detecting various peptitis
  • Detection type gene chip for detecting various peptitis
  • Detection type gene chip for detecting various peptitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] 1. Primer sequence:

[0060] Wherein the HBV primer1 sequence is as follows:

[0061] 5′-GTTTTATTCATATTCCCTCT-3′

[0062] 5′-GGTTTTTATTAGGGTTCAA-3′

[0063] The HBV primer2 sequence is as follows:

[0064] 5′-GAGGCTGTAGGCATAAAT-3′

[0065] 5′-GGGCATTTGGTGGTCTG-3′

[0066] A total of two pairs of primers were designed for HCV amplification primers,

[0067] Wherein the HCV primer1 sequence is as follows:

[0068] HCVi: 5′-GCCTTGTGGTACTGC-3′

[0069] HCVlC: 5'-5ACCGCTCGGAAGTC-3'

[0070] The sequence of HCV primer21 is as follows:

[0071] HCV2: 5′-GGCTTTACCGGCGAC-3′

[0072] HCV2C: 5′-GCTCATACCAiGCAC-3′

[0073] 2. DNA and RNA extraction from serum samples

[0074] Add 100 μl of serum to 10 μl of DEPC-ethanol solution (10% diethylpyrocarbonate (DEPC), 90% absolute ethanol) and let it stand at room temperature for 10 minutes, then in a boiling water bath for 10 minutes, then dry the tube cover at 65°C for 15 minutes. Minutes, centrifuged for 5 minutes (12000 rpm...

Embodiment 2

58.0

CCACACGTCG CGCCTCATTT

[0095] 2. Synthesize the above probe and label the amino group at the 5' end, dilute it with spotting buffer, spot the sample on the glass slide with the modified arm molecule with a spotting instrument, and fix it.

[0096] Serum sample DNA, RNA extraction

[0097] Add 100 μl of serum to 10 μl of DEPC-ethanol solution (10% diethylpyrocarbonate (DEPC), 90% absolute ethanol) and let it stand at room temperature for 10 minutes, then in a boiling water bath for 10 minutes, then dry the tube cover at 65°C for 15 minutes , centrifuged for 5 minutes (12000 rpm, centrifuged for 10 minutes), set aside.

[0098] 3. Prepare template DNA by reverse transcription

[0099] Add the following reagents in sequence to the eppendorf tube containing the DNA and RNA precipitates prepared in step 2:

[0100] Milli-Q H2O (ultrapure water) 11 μl, 15 μmol / L HCV, HCV, reverse transcription primer 2 μl and fully dissolve the precipitate, mix the sample, cen...

Embodiment 3

[0115] 1. Design probes for virus detection of HBV, HCV, HDV, HGV, TTV and other viruses; design probes for detection of subtypes of HBV and HCV; design the following probes for detection of mutation types of HBV viruses .

[0116] 2. Synthesize the above probe and label the amino group at the 5' end, dilute it with spotting buffer, spot the sample on the glass slide with the modified arm molecule with a spotting instrument, and fix it.

[0117] Serum sample DNA, RNA extraction

[0118] Add 100 μl of serum to 10 μl of DEPC-ethanol solution (10% diethylpyrocarbonate (DEPC), 90% absolute ethanol) and let it stand at room temperature for 10 minutes, then in a boiling water bath for 10 minutes, then dry the tube cover at 65°C for 15 minutes. Minutes, centrifuged for 5 minutes (12000 rpm, centrifuged for 10 minutes), set aside.

[0119] 3. Prepare template DNA by reverse transcription

[0120] Add the following reagents in sequence to the eppendorf tube containing the DNA and RN...

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Abstract

The present invention provides a detection gene chip for diagnosing serveral kinds of hepatitis and new primer with high detection rate for using the gene chip in various subtype amplification. The gene chip includes detection quality controlling system and disease diagnosing system. The chip can be used to detect the virus ites of hepatitis B and hepatitis C and detect hepatitis D, hepatitis E, hepatitis G and TTV simultaneously. It has short detection period, high diagnosis accuracy and low diagnosis cost and may be used in epidemiological investigation and blood detection for identifying hepatitis gene type.

Description

technical field [0001] The technical field that the present invention relates to is a gene chip for hepatitis, especially a gene chip for detecting hepatitis mutation. Background technique [0002] Biochip mainly refers to microanalysis units and systems constructed on the surface of solid chips through planar microfabrication technology and supramolecular self-assembly technology. Biochips can integrate many different functional devices: for example, pretreatment of biological samples, extraction of genetic material, amplification of specific gene fragments, biological probe arrays and capillary electrophoresis to form an overall microfluidic system to achieve Accurate, rapid, and informative screening or detection of compounds, proteins, nucleic acids, cells, and other biological components. The gene chip is the most important type of biochip, which integrates a large number of densely arranged gene probes and can analyze a large number of genes in a short time, so that p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 赵伟刘全俊刘伟陆祖宏
Owner 赵伟
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