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Method for detecting necrosis degree of hepatic cells through level of plasma free DNA

A liver cell necrosis and level detection technology, which is applied in the field of plasma free DNA level detection of liver cell necrosis, can solve the problems of other researches that have not been reported in the literature

Inactive Publication Date: 2011-01-26
赵英仁
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some scholars in China have detected the level of plasma nucleosomes in patients with severe hepatitis, and believe that it is higher than that of acute hepatitis, chronic hepatitis and liver cirrhosis, and compared it with ALT. Other studies have not been reported in the literature. Quantitative determination of nucleosomes in severe hepatitis is feasible

Method used

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  • Method for detecting necrosis degree of hepatic cells through level of plasma free DNA
  • Method for detecting necrosis degree of hepatic cells through level of plasma free DNA
  • Method for detecting necrosis degree of hepatic cells through level of plasma free DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1, preparation of primer β-globin standard.

[0027] 1. Main reagents and equipment for the experiment

[0028] 1) Commonly used chemical reagents such as sodium hydroxide, potassium hydroxide, sodium chloride, potassium chloride, absolute ethanol, glacial acetic acid, ammonium acetate, sodium acetate, and isoamyl alcohol are all produced by Xi’an Chemical Reagent Factory, and the purity level is analytical pure;

[0029] 2) EB (subpackaged by Sigma Company, purchased from Huamei Bioengineering Company);

[0030] 3) loading buffer (TaKaRa products of Treasure Biotechnology, purchased from Xi'an Zhou Dingguo Bioengineering Company);

[0031] 4) agar powder (GEGE TECH Shanghai Co., Ltd., purchased from Xi'an Zhou Dingguo Biological Engineering Company);

[0032] 5) Agarose (the original product of Amresco Company, purchased from Xi'an Zhou Dingguo Bioengineering Company);

[0033] 6) Nucleic acid molecular weight reference material 500bp DNA maker, 2000bp DNA ...

Embodiment 2

[0155] Embodiment two, extraction and quantification of plasma nucleosome

[0156] 1. The main materials of the experiment

[0157] Fluorescent PCR reaction mix SYBR Premix Ex Taq TM , with SYBR Green I, reaction buffer, dNTP, TaKaRa Ex Taq TM HS, magnesium ions (TaKaRa product of Bao Biotechnology, purchased from Bao Tech Bioengineering Company), and reagents were stored according to the instructions.

[0158]

[0159] 2. Research object

[0160] A total of 204 patients with acute and chronic liver damage and liver cancer caused by hepatitis B, hepatitis C, and drug-induced hepatitis were admitted to the Department of Infectious Diseases of the First Affiliated Hospital of Xi'an Jiaotong University School of Medicine from March 2009 to June 2010. The clinical diagnosis was in line with the viral hepatitis prevention and treatment program revised by the Xi'an Conference in 2000. Aged 5-79 years old, with an average of 45.14±14.547 years old; 146 males, aged 5-79 year...

Embodiment 3

[0192] Embodiment three, clinical experiment

[0193] 1. Research object

[0194]A total of 204 patients with hepatitis B virus, hepatitis C virus and drug-induced acute and chronic liver damage and liver cancer were admitted to the Department of Infectious Diseases of the First Affiliated Hospital of Xi'an Jiaotong University School of Medicine from March 2009 to June 2010 , to rule out various immune diseases. There is no absolute pathological boundary between clinical diagnosis of chronic hepatitis and compensated liver cirrhosis. Some clinical diagnoses of chronic hepatitis may have shown early liver cirrhosis, both of which are mainly inflammatory reactions; severe hepatitis, acute hepatitis and liver cancer all present Necrosis or apoptosis of liver cells or cancer cells, mainly cell death. The patients were divided into three groups: the first group was CH and LC group, including patients with chronic hepatitis and liver cirrhosis; the second group was DLC group, incl...

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Abstract

The invention relates to a method for detecting the necrosis degree of hepatic cells through the level of plasma free DNA, and belongs to the technical field of biology. In the method, through the separation and extraction method and quantification of the plasma free DNA, the necrosis degree of the hepatic cells is reflected by the level of the free DNA. The method comprises the following steps of: preparing a primer beta-globin serving as a standard substance; extracting the plasma DNA in a plasma specimen of hepatopath; performing a real-time fluorescent quantitative polymerase chain reaction on the plasma DNA extracted in the plasma specimen; and measuring and calculating the concentration of the plasma free DNA. The method provides a new diagnostic index for the diagnosis of severe hepatitis.

Description

technical field [0001] The invention relates to a method for detecting the degree of hepatic cell necrosis at the plasma free DNA level, and belongs to the field of biotechnology. Background technique [0002] Liver disease is a common clinical disease. Only hepatitis B, hepatitis C and other viral hepatitis patients have nearly 500 million people in the world, accounting for 1 / 12 of the global population. China is an area with a high incidence of chronic hepatitis B, and about 350,000 people die of hepatitis B every year. Related diseases, such as liver cirrhosis, liver cancer, severe hepatitis, etc., 1% of hepatitis B patients have severe hepatitis. Severe hepatitis is a type of syndrome of liver failure caused by a large amount of liver necrosis in a short period of time under the action of various pathogenic factors. Its clinical manifestations are characterized by severe disease, rapid development, many complications, and high mortality , is still an urgent problem to ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 赵英仁何英利阎志李倩崔美灵王科陈天艳刘红莉
Owner 赵英仁
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