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Method for constructing detecting system of hepatitis type B virus DNA

A technology of magnetic nanoparticles and marker genes, which is applied in the field of detection of viral DNA and its mutation, to reduce steric hindrance, ensure hybridization rate, and improve specific selectivity

Inactive Publication Date: 2005-09-14
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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Problems solved by technology

At present, there is no literature that discloses the use of Fe 3 o 4 (Core) / Gold (Shell) Nanoparticle Virus DNA Probe, Method for Detecting Virus DNA and Its Variation Through Secondary Amplification Technology

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  • Method for constructing detecting system of hepatitis type B virus DNA

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example 1

[0015] Example 1, Fe 3 o 4 (Core) / Au(Shell) Nanoparticle HBV DNA Probe for Ultrasensitive Detection of HBV DNA

[0016] Oligonucleotide design and synthesis:

[0017] The oligonucleotide probe consists of 26 nucleotides and consists of two parts, the main part is complementary to the mutation region or conserved region of HBV DNA, and is 20 nucleotides long; the other small part is 6 nucleotides acid T to ensure that the main part of the oligonucleotide is fully combined with its complementary region.

[0018] 1. Screening, cloning and purification of variant strains and wild strains of various types of hepatitis B HBV DNA (acute, chronic / severe, mild and virus carriers). Find out the mutation regions related to the occurrence and development of hepatitis B and conduct systematic research, design probes for common mutation regions; at the same time design probes for HBV DNA conserved sequence regions. Design probes with HBV DNA (accession: X75658) 1873-1912 as the target s...

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Abstract

A process for creating a detection system of HBV DNA includes such steps as designing oligonucleotide probe by using the 1873-1912 sites of HBV DNA as target sequence, dividing the probe into two parts (one including 20 nucleotides and another including 6 nucleotides), using the paraffin thiol to decorate the 5' or 3' terminal of oligonucleotide, using Au-S bone to link to probe to Au or Fe3O4 / Au nanoparticle, using manipulated capture probe, and adding linking chain and signal amplifying substances in the detection system. Its advantages are high sensitivity and specificity and low cost.

Description

technical field [0001] The invention relates to the technical field of virus gene detection. Specifically, the use of Fe 3 o 4 (Core) / gold (shell) nanoparticle virus deoxyribonucleic acid (deoxyribonucleic acid, DNA) probe, through secondary amplification technology, to establish a method for detecting virus DNA and its mutation. Background technique [0002] Nanoparticles are particles with a diameter of 1-100nm. In this scale range, nanomaterials have many new properties, so it has become a hot field of scientific research. Utilizing the unique characteristics of force, electricity, light, and magnetism of nanomaterials, a highly interdisciplinary nanobiomedical technology has been formed, which has a wide range of application values ​​in reality. In 1996, Chad A Mirkin et al covalently bound alkanethiol-modified oligonucleotides to the surface of gold nanoparticles through gold-sulfur (Au-S) bonds to form DNA probes labeled with gold nanoparticles, and Using this prob...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 宁琴姚凯伦罗小平刘祖黎习东卢强华
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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