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A method for rapid detection of hepatitis B circular dna

A technology of DNA molecules and probes, which is applied in the field of enrichment, separation and rapid detection of hepatitis B circular DNA, can solve the problems of unsatisfactory specificity and sensitivity, and achieve the effect of high sensitivity, good application prospects, efficient and accurate detection

Active Publication Date: 2020-01-17
THE AFFILIATED HOSPITAL OF TRADITIONAL CHINESE MEDICAL TO SOUTHWEST MEDICAL UNIV
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Problems solved by technology

However, the specificity and sensitivity of these methods are not ideal

Method used

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  • A method for rapid detection of hepatitis B circular dna
  • A method for rapid detection of hepatitis B circular dna

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Experimental program
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Effect test

experiment example 1

[0035] Experimental Example 1 The present invention detects the method for HBV cccDNA

[0036] 1. Detection method

[0037] 1. Preparation of HBV cccDNA-specific DNA molecular probe magnetic nanospheres

[0038] Preparation process such as figure 1 Shown:

[0039] 1. Preparation of HBV cccDNA-specific DNA molecular probe magnetic nanospheres

[0040] 1.1 Preparation of Fe by chemical coprecipitation method 3 o 4 Nanoparticles:

[0041] 20ml FeCl 3 (1.0mol / L) with 5ml FeCl 2(2.0mol / L, prepared in 2.0mol / L hydrochloric acid solution) the solution is mixed evenly and added to 250ml 0.7mol / L ammonia solution, and the dark brown precipitate obtained after centrifugation is mixed with 150ml 2.0mol / L perchloric acid The solution was dispersed, washed with ultrapure water until neutral, and dried to obtain Fe 3 o 4 Nanoparticles.

[0042] 1.2 Preparation of silicon-shell magnetic nanoparticles by inverse emulsion method:

[0043] Mix Triton X-100, n-hexanol and cyclohexane...

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Abstract

The invention discloses a method of rapidly detecting hepatitis B circular DNA. The method of rapidly detecting HBV cccDNA comprises the following steps: (I) adopting an HBV cccDNA specificity DNA molecular probe magnetic nano microsphere to separate and enrich HBV cccDNA; (II) amplifying and purifying HBV cccDNA; and (III) carrying out rapid detection. The HBV cccDNA specificity DNA molecular probe magnetic nano microsphere is prepared by the following steps: (1) adopting a chemical co-precipitation method to prepare Fe3O4 nano particles; (2) adopting inverse emulsion to prepare a silicon shell magnetic nano particle; (3) modifying the silicon shell magnetic nano particle by avidin; and (4) coupling modified magnetic nano particle with an ssDNA probe aiming at specific sequence of cccDNA.The provided method can effectively and rapidly detect HBV cccDNA, has high specificity and sensitivity, and has a good application prospect in clinic.

Description

technical field [0001] The invention relates to a method for enriching, separating and rapidly detecting hepatitis B circular DNA. Background technique [0002] Chronic hepatitis B is a disease caused by infection with hepatitis B virus (HBV), and there is currently no specific drug that can cure it. HBV cccDNA is the key factor for the persistence of hepatitis B virus infection and is difficult to cure. By detecting HBV cccDNA, we can understand the status and infectivity of HBV infection, evaluate the efficacy of drug treatment of HBV, help understand the degree of HBV clearance from the body, and judge when to stop for clinicians Whether HBV will actively re-replicate and lead to hepatitis B recurrence after antiviral treatment and cessation of treatment provides an objective indicator, and it is also an indicator to evaluate whether extrahepatic tissues are infected by HBV and whether the transplanted liver is reinfected in liver transplantation. [0003] Scholars at ho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/26
CPCG01N27/26G01N27/3278
Inventor 郭永灿涂植光赵朝辉王友强
Owner THE AFFILIATED HOSPITAL OF TRADITIONAL CHINESE MEDICAL TO SOUTHWEST MEDICAL UNIV
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