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Magnetic biological probe and test strip for detecting hepatitis B virus (HBV) and preparation method and using method of biological probe

A biological probe and detection line technology, applied in the field of biotechnology and nanomaterials, can solve the problems of "two pairs of half" reagents that cannot well reflect the existence of hepatitis B virus, high cost of detection experiments, and inability to quantitatively detect HBV.

Inactive Publication Date: 2013-09-11
上海爱纳玛斯医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem solved by the present invention is to overcome the defects that the "two-and-a-half" reagents cannot well reflect the existence of hepatitis B virus, cannot quantitatively detect HBV, and the HBV-DNA quantitative PCR detection experiment is high in cost and time-consuming, and provides a A magnetic nanoparticle biological probe for hepatitis B virus detection and its preparation method, and a magnetic immunochromatographic test strip for HBV detection, its preparation method and use method

Method used

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  • Magnetic biological probe and test strip for detecting hepatitis B virus (HBV) and preparation method and using method of biological probe
  • Magnetic biological probe and test strip for detecting hepatitis B virus (HBV) and preparation method and using method of biological probe
  • Magnetic biological probe and test strip for detecting hepatitis B virus (HBV) and preparation method and using method of biological probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1 magnetic nanoparticles

[0061] A method for preparing magnetic nanoparticles, comprising the steps of:

[0062] 1) Preparation of magnetic particles by solvothermal method: 0.85g FeCl 3 ·6H 2 O and 0.4033g PAA were respectively dissolved in 40ml of ethylene glycol, and then 1.2g of urea was added to dissolve. After mixing, they were added to a stainless steel reactor with a tetrafluoroethylene liner, tightened the reactor, and heated at 220°C for 16 hours. With the assistance of magnetic separation, wash with ethanol and deionized water three times each to obtain magnetic particles with an average particle size of 90 nm;

[0063] 2) Silicon oxide-coated magnetic particles: Mix 90 mg of magnetic particles with 1.0 mol / L hydrochloric acid solution, sonicate for 30 minutes, wash with deionized water for 5 times with the assistance of magnetic separation, and then mix the magnetic particles with alcohol-water volume ratio Mix 125mL of a 70:30 alcohol-water ...

Embodiment 2

[0065] Embodiment 2 magnetic nanoparticles

[0066] A method for preparing magnetic nanoparticles, comprising the steps of:

[0067] 1) Preparation of magnetic particles by solvothermal method: 0.81g FeCl 3 ·6H 2 O and 2.66g of sodium citrate were dissolved in 30ml of ethylene glycol respectively, mixed and added to a stainless steel reactor with a tetrafluoroethylene liner, tightened the reactor, heated and reacted at 200°C for 24 hours, and then assisted by magnetic separation , washed three times with ethanol and deionized water respectively, to obtain magnetic particles with an average particle diameter of 250nm;

[0068] 2) Silicon oxide-coated magnetic particles: Mix 110 mg of magnetic particles with 1.0 mol / L hydrochloric acid solution, sonicate for 15 minutes, wash with deionized water for 7 times with the assistance of magnetic separation, and then mix the magnetic particles with alcohol-water volume ratio Mix 125 mL of 90:10 alcohol-water mixture, add 100 μL of te...

Embodiment 3

[0070] Embodiment 3 is used for the magnetic nanoparticle bioprobe that HBV detects

[0071] The preparation method of the magnetic nanoparticle biological probe that is used for HBV detection, it comprises the steps:

[0072] (1) Add 15 mg of the magnetic nanoparticles prepared in Example 1 or 2 to 50 g of ethanol aqueous solution with a volume ratio of alcohol to water of 70:30, add 50 μL of ammonia water and 35 μL of aminopropyltriethoxysilane, and stir for 2 hours at room temperature , continue to stir for 3 hours in a water bath at 70°C, wash with ethanol to obtain surface-aminated magnetic nanoparticles; mix the surface-aminated magnetic nanoparticles with dimethyl sulfoxide (DMSO), add 5.5 mg of succinic anhydride to react for 24 hours , washed three times with DMSO under the assistance of magnetic separation, and then washed three times with deionized water to obtain surface carboxylated magnetic nanoparticles, the carboxyl content of these surface carboxylated magneti...

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Abstract

The invention discloses a magnetic nanoparticle biological probe and a test strip for detecting hepatitis B virus (HBV) and a preparation method and a using method of the biological probe. The method for preparing the biological probe comprises the following steps of: performing surface amino functionlization treatment on magnetic nanoparticles with the particle size of 50-300nm, performing carboxylation treatment, mixing and reacting the magnetic nanoparticles subjected to surface carboxylation, carbodiimide and N-hydroxy succinimide in a buffer solution, and washing to obtain a reactant A; mixing and reacting the reactant A and an antibody for a HBV marker in a coupled buffer solution to obtain a reactant B; mixing and reacting the reactant B and a solution of a compound containing amino, and washing to obtain the product. The biological probe is high in specificity, sensitive in signal and high in stability and is suitable for detecting multiple hepatitis B markers according to the selected targeting markers and matched antibodies. According to the test strip, an obtained magnetic signal can be subjected to secondary determination through visual inspection and MAR detection, the personal error can be greatly reduced, and the accuracy is high.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and nanomaterials, in particular to a magnetic nanoparticle biological probe for detecting hepatitis B virus, a magnetic immunochromatographic test strip, and a preparation method and application method thereof. Background technique [0002] Hepatitis B virus, referred to as hepatitis B, is a disease caused by the infection of the body by the hepatitis B virus (HBV for short). Hepatitis B virus is a hepatotropic virus that mainly exists in liver cells and damages liver cells, causing inflammation, necrosis, and fibrosis of liver cells. According to the report of the World Health Organization, about 2 billion people in the world have been infected with HBV, of which 350 million people are chronically infected, and about 1 million people die every year from liver failure, liver cirrhosis and primary liver cancer (HCC) caused by HBV infection. ). The results of the hepatitis B epidemic virus...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/576
Inventor 崔正权
Owner 上海爱纳玛斯医药科技有限公司
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