Method for detecting hepatitis b virus subtype based on DNA paper-folding mark

A hepatitis B virus, virus technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of lack of visual labels in genetic information analysis

Pending Publication Date: 2019-08-02
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the resolution of AFM can achieve sub-10nm level, its application in genetic information analysis is still limited by the lack of unique visual labels

Method used

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  • Method for detecting hepatitis b virus subtype based on DNA paper-folding mark
  • Method for detecting hepatitis b virus subtype based on DNA paper-folding mark
  • Method for detecting hepatitis b virus subtype based on DNA paper-folding mark

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Embodiment 1, the kit that detects different genotypes of virus

[0090] figure 1 It is a schematic diagram of directly reading DNA origami markers to identify HBV genotypes based on AFM imaging, specifically: HBV genomic DNA is extracted from serum samples, amplified by asymmetric PCR and digested with λ exonuclease, and the HBV target genome fragments are used with the corresponding DNA origami markers are used as "amplified signals" for site-specific labeling, and the corresponding markers are read directly on the AFM image to translate genotype information.

[0091] 1. Kits for detecting different genotypes of viruses

[0092] Kits for detecting different genotypes of viruses, including multiple sets of biological products;

[0093] Each set of biological products includes one or more groups of reagents;

[0094] Each set of biological products corresponds to 1 virus genotype and 1 shape of DNA origami;

[0095] Each group of reagents consists of a DNA origami w...

Embodiment 2

[0129] Embodiment 2, the application of triangle DNA origami and cross DNA origami in detecting HBV genotype

[0130] Figure 6 It is a schematic diagram of the structure of the DNA origami label and the position of the probe extension used in the experiment.

[0131] 1. Detection of HBV genotype by triangle DNA origami and cross DNA origami

[0132] 1. Extraction of total nucleic acid from the sample to be tested

[0133] The samples in this study came from the whole blood samples of patients who were diagnosed with HBV infection by ELISA immune reaction pre-detection in Wuxi People's Hospital, of which 5 cases were pre-diagnosed as genotype B, and the other 6 cases were genotype C. Use the TIANampVirus DNA / RNAKit Viral Genome Nucleic Acid Extraction Kit to extract HBV viral DNA from whole blood samples.

[0134] The DNA was eluted in 50 μl of elution buffer (included with the kit), stored at -80° C. for future use, and the HBV viral DNA of 11 samples was obtained.

[013...

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Abstract

The invention discloses a method for detecting a hepatitis b virus subtype based on a DNA paper-folding mark. A kit provided by the invention includes multiple sets of biological products; each set ofbiological products includes one or multiple sets of reagents; each set of biological products is corresponding to one virus genotype and a DNA folding paper of one shape; each set of reagents is composed of a DNA folding paper with an extending area and a mediate probe corresponding to the DNA folding paper; the extending area of the DNA folding paper in each set of reagents is a free area in size of 54-1119nt extending after 5 Ts extending from one short chain at each endpoint of the DNA folding paper. The invention develops a set of self-assembling method-based DNA folding paper nanometerstructure which is used as a specific mark. A specifically designed probe is used for capturing and marking a clinic hepatitis B virus sample DNA and finally recognizing a corresponding HBV genotype from atomic force microscope (AFM) scanning.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting hepatitis B virus subtypes based on DNA origami markers. Background technique [0002] Chronic hepatitis B virus (HBV) infection is a serious public epidemic, which may evolve into liver cirrhosis and hepatocellular carcinoma (HCC) without proper diagnosis and treatment measures, and its harm spreads globally and potentially huge health harm. Existing methods for detecting HBV infection mainly rely on serological and molecular testing. Serological testing identifies hepatitis B surface antigen (HBsAg) as an important diagnostic marker in patient sera by identifying virus-encoded antigens and their corresponding antibodies, usually by enzyme immunoassay analysis. Although commercial standard HBV antigen assays have been widely used in clinical applications, detection limits that are highly dependent on viral antigen concentration may lead to erroneous results ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70
CPCC12Q1/706C12Q1/707C12Q1/6816C12Q2563/155
Inventor 师咏勇潘敦刘轲樊春海贺林
Owner SHANGHAI JIAO TONG UNIV
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