Primers, probes and kits for predicting liver cancer caused by hepatitis B virus

A technology of hepatitis B virus and hepatitis B virus, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve problems such as no uniform standard, small sample size, and single mutation site , to achieve the effect of fast detection speed, high sensitivity and strong selectivity

Active Publication Date: 2011-12-14
AMOYDX BIOTECHNOLOGY RES CENT CO LTD
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Problems solved by technology

[0011] Although many researchers have done research on HBV-related mutations in HCC patients, these studies generally have small sample sizes, and the reported mutation sites are relatively single. Which mutation sites can be used as mutation points for early prediction and diagnosis of HCC remains to be seen. no uniform standard

Method used

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  • Primers, probes and kits for predicting liver cancer caused by hepatitis B virus
  • Primers, probes and kits for predicting liver cancer caused by hepatitis B virus
  • Primers, probes and kits for predicting liver cancer caused by hepatitis B virus

Examples

Experimental program
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Effect test

Embodiment 1

[0041] According to the sequences of the hepatitis B gene and each mutation point, primers and probes were designed using the Primer Premier 5.0 software of Canada's Premier Company. The primer sequences are shown in Table 1.

[0042] A total of 150 serum samples of HCC patients and 150 serum samples of non-HCC and non-LC hepatitis B patients were collected in this experiment as the control group. First, take 100 μL of the patient's serum sample, and extract the DNA of the hepatitis B virus according to the instructions of the hepatitis B virus detection kit (Xiamen Aide Biomedical Technology Co., Ltd.). Take 5 μL of the DNA to be tested as a template, and use real-time fluorescent PCR for amplification. The conditions for PCR amplification are as follows:

[0043] Real-time fluorescent PCR 50μL reaction system:

[0044]

[0045] Real-time PCR reaction conditions are pre-denaturation at 95°C for 3 minutes, 10 cycles: denaturation at 95°C for 20 seconds, annealing at 62°C f...

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Abstract

The invention discloses a primer, a probe and a kit used for predicting liver cancer caused by hepatitis B virus (HBV). The primer and the probe are designed according to mutation sites of HBV. The mutation sites comprise the following 14: C1653T, T1674C, T1753V, A1762T, G1764A, A1846T, A1858T, G1896A, G1899A, C1485T, C1499H, G1613A, C3116T and T53C. According to the invention, early stage prediction of hepatocellular carcinoma is realized through detections upon the 14 mutation sites of HBV. Also, the invention provides a method for detecting HBV mutation sites.

Description

technical field [0001] The present invention relates to biological reagents, in particular to a detection method for predicting hepatitis B virus-related liver cancer, in particular to hepatitis B virus mutations (C1653T, T1674C, T1753V, A1762T, G1764A, A1846T, A1858T, G1896A, G1899A, C1485T , C1499H, G1613A, C3116T and T53C) detection. Background technique [0002] Hepatitis B virus (HBV) infection has become a global problem that seriously threatens human health. About 3 billion people in the world have ever been infected with hepatitis B virus, accounting for about half of the world's total population. Nearly 350 million people are carriers of HBV (Ganem D, Prince AM. Hepatitis B virus infection-natural history and clinical consequences. N Eng J Med 2004; 350: 1118-1129). HBV infection is one of the most important risk factors for hepatocellular carcinoma (HCC) and liver cirrhosis (LC). Epidemiological surveys show that 80% of HCC patients in the world are caused by HB...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
Inventor 王蓟斌张海龙阮力曾骥孟郑立谋
Owner AMOYDX BIOTECHNOLOGY RES CENT CO LTD
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