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Kit for directly observing distribution and location of hepatitis B virus (HBV) cccDNA in liver tissue

A liver tissue, direct technology, applied in the field of in situ detection of hepatitis B virus cccDNA, can solve the problems of HBVcccDNA distribution relationship, cannot reflect the relationship of HBVcccDNA, cannot observe the distribution and location of HBVcccDNA, etc., and achieve the effect of high sensitivity and simple method

Active Publication Date: 2012-07-04
钟彦伟
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  • Description
  • Claims
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Problems solved by technology

[0011] In addition, this method is a specific method for quantitatively detecting HBVcccDNA in liver tissue, but it cannot relate the distribution of HBVcccDNA to liver tissue morphology or histopathological characteristics, cannot reflect the relationship between HBVcccDNA and liver tissue structure, and cannot observe the distribution of HBVcccDNA in liver tissue. Intracellular distribution and localization

Method used

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  • Kit for directly observing distribution and location of hepatitis B virus (HBV) cccDNA in liver tissue
  • Kit for directly observing distribution and location of hepatitis B virus (HBV) cccDNA in liver tissue
  • Kit for directly observing distribution and location of hepatitis B virus (HBV) cccDNA in liver tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1 directly observes the detection method of HBVcccDNA distribution and localization in liver tissue

[0059] 1. Research samples

[0060] The research subjects were 50 patients with chronic hepatitis B, liver cirrhosis, and hepatocellular carcinoma from March 2010 to May 2011 in the 302 Hospital of the People's Liberation Army, including 34 males and 16 females, ranging in age from 7 to 68 years old. The diagnosis was in line with the diagnostic criteria for viral hepatitis revised by the Tenth National Viral Hepatitis Conference in 2000, excluding hepatitis A, C, D, E, and G viruses, transfusion-transmitted viruses, cytomegalovirus, Epstein-Barr virus infection, and alcoholic and drug-related infections. Sexual, autoimmune liver disease. Liver tissue samples from non-hepatitis B patients were also collected as negative controls.

[0061] 2. Method

[0062] 1. Processing of tissue slices

[0063] Put the liver biopsy tissue from the patient into 10% forma...

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Abstract

The invention relates to a method and a kit for detecting hepatitis B virus cccDNA in situ. The method comprises the following steps of: fixing a sample section of a liver tissue; digesting; performing in-situ rolling cycle and fluorescence-quantitation polymerase chain reaction (PCR) amplification; and detecting an amplification product. By the method, the distribution and location of the hepatitis B virus (HBV) cccDNA in the liver tissue can be directly observed, the operation is simple and convenient, the liver tissue is not required to be made into homogenate, nucleic acid is not required to be extracted to serve as a template, and judgment results are not required to be made into a standard curve.

Description

Technical field: [0001] The invention relates to a method for in-situ detection of hepatitis B virus cccDNA, in particular to a method and kit for directly observing the distribution and location of HBVcccDNA in liver tissue. Background technique: [0002] The detection of hepatitis B virus covalently closed circular DNA (covalently closed circular DNA, hereinafter referred to as HBVcccDNA) is of great significance in the study of the pathogenesis of viral diseases and antiviral therapy. [0003] At present, there are literatures using the nested PCR method to detect HBV cccDNA expression in situ in liver tissue. This method cannot exclude the interference of non-closed circular DNA and integrated HBV DNA. [0004] PCR technology can amplify a small amount of nucleic acid several million times in a reaction tube for analysis, but it cannot reflect the relationship between the amplification product and the tissue structure. Although the in situ hybridization technique can re...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 钟彦伟
Owner 钟彦伟
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