Kit and method for testing hypotype of virogene of hepatitis B and drug fast mutation

A technology of hepatitis B virus and drug-resistant mutations, applied in biochemical equipment and methods, microbe measurement/testing, etc., can solve the problems of increasing the economic burden of patients and failing to detect HBV genotype subtypes, so as to reduce the economic burden Effect

Active Publication Date: 2007-12-26
亚能生物技术(深圳)有限公司
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  • Abstract
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Problems solved by technology

[0003] Chinese patents 200410025199.X and 200510065445.9 respectively disclose a method for detecting lamivudine-resistant HBV DNA, but this method cannot detect HBV genotypes while measuring drug resistance
So far, there have been no products and reports for simultaneous detection of HBV genotyping and resistance to two commonly used drugs at home and abroad. Therefore, if comprehensive reference information is required clinically, patients need to do multiple tests, which greatly increases the financial burden on patients.

Method used

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  • Kit and method for testing hypotype of virogene of hepatitis B and drug fast mutation
  • Kit and method for testing hypotype of virogene of hepatitis B and drug fast mutation
  • Kit and method for testing hypotype of virogene of hepatitis B and drug fast mutation

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Embodiment 1

[0051] 1. Materials

[0052] 1. Reagents:

[0053] 1) 3%H 2 O 2

[0054]2) 20×SSC (pH 7.0): NaCl 175.3g

[0055] Sodium Citrate 88.2g

[0056] Add 750 mL of distilled water to dissolve, adjust the pH to 7.0 with a pH meter, and finally dilute to 1000 mL, and store in autoclave.

[0057] 3) 10% SDS (pH 7.0): SDS 20g

[0058] Add 180 mL of distilled water to dissolve, adjust the pH to 7.0 with a pH meter, and finally set the volume to 200 mL. 1M sodium citrate (pH5.0): sodium citrate 294g

[0059] Add 700 mL of distilled water to dissolve, adjust the pH to 5.0 with concentrated HCl, and finally set the volume to 1000 mL.

[0060] 4) Solution A (2×SSC, 0.1% SDS, pH7.4): 20×SSC 100mL

[0061] 10% SDS 10mL

[0062] Add distilled water to make up to 1000mL.

[0063] 5) Solution B (0.5×SSC, 0.1% SDS, pH7.4): 20×SSC 25mL

[0064] 10% SDS 10mL

[0065] Add distilled water to make up to 1...

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Abstract

This invention provides method and test kit for synchronously detecting sub-genotypes and drug-resistant mutation of hepatitis B virus (HBV). The test kit comprises: two specific primers for amplifying gene or transcript of HBV, three specific oligonucleotide probes for detecting sub-genotypes of HBV, and thirteen specific oligonucleotide probes. The primers and the probes can also be replaced with the complementary sequences. The test kit can synchronously differentiate three sub-genotypes (B, C and D) of HBV, and detect lamivudine- and adefovir dipivoxil-resistant mutations. The test kit can reduce the detection cost.

Description

technical field [0001] The invention relates to the field of virus subtype and drug resistance mutation detection, in particular to a kit and detection method for simultaneously detecting hepatitis B virus gene subtype and drug resistance mutation. Background technique [0002] According to the World Health Organization, there are about 350 million people in the world with hepatitis B virus, and about 220 million people in Asia. my country is the most important hepatitis B country. In 2002, my country's epidemiological survey showed that the positive rate of hepatitis B surface antigen was 9.09%, about 120 million, and the number of chronic hepatitis B patients was about 30 million. At present, this group of people is in the peak period of incidence and treatment. HBV infection is prevalent worldwide, and its genotype distribution has regional characteristics. In my country and Asia, B and C types are the main types, and there is a small amount of D type. At present, many ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor TIAN JIEREN WEIXIANG ZHULIAO SHENGYUNZOU YAODONG
Owner 亚能生物技术(深圳)有限公司
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