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Ultra-sensitive magnetic nanometer detection technology for hepatitis C virus (HCV)

A superparamagnetic and nanosphere technology, applied in the direction of nanotechnology, nanotechnology, nanotechnology for sensing, etc., can solve the problems of non-applicable application of toxicity, change of magnetic properties, etc., and achieve specificity improvement and detection sensitivity Improvement, accurate and reliable results

Inactive Publication Date: 2017-02-01
北京国际旅行卫生保健中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nanoparticles of metals and alloys have good structural and magnetic properties, but their magnetic properties will be changed due to the oxidation of metals and alloys. In addition, their obvious toxicity also makes them unsuitable for in vivo applications.

Method used

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  • Ultra-sensitive magnetic nanometer detection technology for hepatitis C virus (HCV)
  • Ultra-sensitive magnetic nanometer detection technology for hepatitis C virus (HCV)
  • Ultra-sensitive magnetic nanometer detection technology for hepatitis C virus (HCV)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Detection of microRNA expression in viruses

[0050] 1. Extraction of serum viral DNA by magnetic bead method

[0051] 1. Add 200ul HCV serum samples to 1.5ml EP tubes, then add 20ul proteinase K (10mg / ml) and 40ul lysate [50mM Tris-HCl (pH8.0), 2.5% SDS, 2.5% TritonX-100 etc.] and mix well.

[0052] Note: add yeast tRNA to the lysate in advance (add 4ug yeast tRNA to each reaction)

[0053] 2. Add 400ul binding solution [5M GuSCN, 20mM Tris-HCl (pH6.6), 10% ethanol, etc.] and 25ul siliconized magnetic beads to the reaction tube, vortex and incubate at room temperature for 3 minutes.

[0054] 3. Place the reaction tube on the magnetic stand for about 30 seconds, and use a pipette to remove the supernatant.

[0055] 4. Add 750ul washing solution [20mM NaCI, 2mM Tris-HCl (pH7.5), 50% ethanol] to the tube, mix well and place on the magnetic stand, discard the supernatant.

[0056] 5. Repeat 4.

[0057] 6. Add 50ul of eluent [10mM Tris-HCl (pH8.0)] to suspend...

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Abstract

The invention discloses a method for surface functionalization treatment of a Fe3O4 nanoparticle and constructs a novel, simple, cheap, convenient and highly-sensitive HCV virus detection technology through coupling of produced active groups and a HCV antibody. According to the invention, a superparamagnetic polymethyl acrylate microsphere is synthesized by using suspension emulsion polymerization method; the surface of the microsphere is modified with a carboxyl group and coupled with an anti-HCV antibody; and the coupled superparamagnetic polymethyl acrylate microsphere has HCV enrichment efficiency of 94.50%. The invention provides a method for detecting HCV by using a combination of immunomagnetic beads and fluorescent PCR, belonging to the field of health quarantine.

Description

technical field [0001] The present invention relates to the significant demand in the detection of hepatitis C virus, using magnetic Fe 3 o 4 Nanospheres use methods such as surface polymer chemical reactions to construct superparamagnetic polymer nanospheres and functionalize the surface of the microspheres to generate active groups, and biomodify the surface of the microspheres by coupling with hepatitis C (HCV) antibodies , Constructed a simple, cheap, convenient high-sensitivity new HCV virus detection technology in serum. It belongs to the field of inspection and quarantine. Background technique [0002] Hepatitis C is a global infectious disease caused by hepatitis C virus (HCV), which affects about 1% of the general population worldwide. Chronic infection of hepatitis C virus (HCV) can lead to chronic inflammation, necrosis and fibrosis of the liver, and some patients may develop liver cirrhosis or even hepatocellular carcinoma (HCC), which is extremely harmful to ...

Claims

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Application Information

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IPC IPC(8): C01G49/08C07K16/10C12N15/10C12Q1/70C12Q1/68B82Y15/00C12R1/93
CPCC01G49/08B82Y15/00C07K16/109C12Q1/6806C12Q1/6851C12Q1/707C12Q2523/308C12Q2563/143C12Q2563/155C12Q2531/113C12Q2561/101C12Q2545/113C12Q2547/101
Inventor 曹姗姗汪琳郭敏卓张文玲周鲁林肖利力王瑾李希红赵相鹏尹羿
Owner 北京国际旅行卫生保健中心
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