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Immunocapture molecule detection method for complete HBV particles

A virus particle and immune capture technology, applied in the field of molecular biology, can solve the problem of little research on virus particle detection

Pending Publication Date: 2021-08-31
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, little research has been done on the detection of intact HBV virions

Method used

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  • Immunocapture molecule detection method for complete HBV particles
  • Immunocapture molecule detection method for complete HBV particles
  • Immunocapture molecule detection method for complete HBV particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Antibody-magnetic bead coupling

[0037] (1) Vigorously shake the carboxylated magnetic beads to disperse them evenly. Take 3.3mg of magnetic beads into a 2mL EP tube, separate by magnetic force, and wash 3 times with pre-cooled MES buffer;

[0038] (2) Apply a magnetic field, remove the supernatant, quickly add 100 μL of NHS and an equal amount of EDC solution to the EP tube, shake vigorously, and activate the magnetic beads continuously at 25°C for 30 minutes;

[0039] (3) With the help of a magnetic stand, wash the magnetic beads 3 times with pre-cooled MES solution, and use them for antibody coupling as soon as possible;

[0040] (4) Dilute the antibody to be coupled with pre-cooled MES solution to a final antibody concentration of about 0.6g / L, 100μL;

[0041] (5) Resuspend the activated magnetic beads in 100 μL of MES solution and shake vigorously to ensure that the magnetic beads are completely dispersed;

[0042] (6) Take 20 μL of activated magnetic bead susp...

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Abstract

The invention provides an immunocapture molecular detection method for complete HBV virus particles. The immunocapture molecular detection method comprises the following steps that a specific antibody is coupled with carboxyl magnetic beads serving as a medium, virus particles are captured and separated, and then QPCR detection is carried out. Experiments show that the virus particles in the sample can be successfully captured and separated, and the virus particles with different components in the sample can be distinguished due to the difference of the magnetic bead coupling antibodies. By means of the immunocapture molecular detection method for the complete HBV virus particles, it is accidentally found that virion components in cell supernatant and serum are different, HBV DNA in the cell supernatant is mainly derived from capsid viruses, and the content of complete virion is small; and the capsid virus content in the serum specimen is lower than that of the complete virus particles. Along with the increase of the HBV DNA copy number in the serum, the content of the complete virus particles in the serum is obviously increased, which prompts that the detection of the complete virus particles can be used as a new serum marker.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to HBV complete virus particles, in particular to an immune capture molecular detection method of HBV complete virus particles. Background technique [0002] Hepatitis B virus (HBV) is a pseudoretrovirus with a double-stranded DNA genome. Its important feature is that HBV can secrete various forms of virus particles after infecting liver cells, which can be roughly divided into virus particles containing genome and genome-less virus particles. According to the type of genome, it can be divided into HBV RNA-like virions, HBV DNA virions and non-genome virions. Based on the presence or absence of hepatitis B surface antigen (HBsAg) as the standard, virions with different genomes can be divided into nucleocapsid viruses, enveloped viruses and subvirions. The most representative ones are subviral particles, NCs (nucleocapsid) particles, HBV RNA virions and complete virions (HBV Dane particl...

Claims

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Application Information

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IPC IPC(8): C12Q1/6804C12Q1/6851C12Q1/70C12R1/93
CPCC12Q1/6804C12Q1/6851C12Q1/706C12Q2563/131C12Q2563/143C12Q2563/149C12Q2531/113C12Q2563/107C12Q2545/113Y02A50/30
Inventor 汪德强刘俊叶黄爱龙伍晓莉邹春红师悦嫄毛胜蓝王雯
Owner CHONGQING MEDICAL UNIVERSITY
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